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Leonurine is a natural product unique to the Lamiaceae plant Leonurus japonicus Houtt., and it has attracted attention due to its anti-oxidative stress, anti-apoptosis, anti-fibrosis, and metabolic regulation properties. Also, it plays an important role in the prevention and treatment of nonalcoholic fatty liver disease (NAFLD) through a variety of biological mechanisms, but its mechanism of action remains to be elucidated. Therefore, this study aims to preliminarily explore the mechanisms of action of leonurine in NAFLD. Mice were randomly divided into four groups: the normal control (NC) group, the Model (M) group, the leonurine treatment (LH) group, and the fenofibrate treatment (FB) group. The NAFLD model was induced by a high-fat high-sugar diet (HFHSD) for 12 weeks, and liver pathological changes and biochemical indices were observed after 12 weeks. Transcriptomic analysis results indicated that leonurine intervention reversed the high-fat high-sugar diet-induced changes in lipid metabolism-related genes such as stearoyl-CoA desaturase 1 (Scd1), Spermine Synthase (Sms), AP-1 Transcription Factor Subunit (Fos), Oxysterol Binding Protein Like 5 (Osbpl5), and FK506 binding protein 5 (Fkbp5) in liver tissues. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis results suggest that leonurine may exert its lipid-lowering effects through the AMP-activated protein kinase (AMPK) signaling pathway. Liver lipidomic analysis showed that leonurine could alter the abundance of lipid molecules related to fatty acyl (FAs) and glycerophospholipids (GPs) such as TxB3, carnitine C12-OH, carnitine C18:1-OH, and LPC (20:3/0:0). Molecular biology experiments and molecular docking techniques verified that leonurine might improve hepatic lipid metabolism through the alpha-1A adrenergic receptor (ADRA1a)/AMPK/SCD1 axis. In summary, the present study explored the mechanism by which leonurine ameliorated NAFLD by inhibiting hepatic lipid synthesis via the ADRA1a/AMPK/SCD1 axis. [ABSTRACT FROM AUTHOR]

BACKGROUND: Leonurine has many biological activities such as improving microcirculation, anti-oxidation, anti-apoptosis, scavenging free radicals, anti- inflammation, and anti-fibrosis, and can promote osteogenic differentiation of bone marrow mesenchymal stem cells, which has the potential to be applied in the treatment of periodontitis. OBJECTIVE: To explore the anti-inflammatory and osteogenic effects of leonurine loading into chitosan/sodium glycerophosphate/sodium alginate hydrogel. METHODS: (1) Chitosan/sodium glycerophosphate/sodium alginate hydrogel (blank hydrogel) and chitosan/sodium glycerophosphate/sodium alginate/leonurus alkali hydrogel were prepared respectively. RAW 264.7 and MC3T3-E1 cells were inoculated with the two kinds of hydrogel. The cytotoxicity of hydrogels was detected by CCK-8 assay and live/dead cell staining. (2) RAW 264.7 cells were cultured in five groups. The blank group was cultured for 24 hours routinely. The lipopolysaccharide group was treated with lipopolysaccharide. The simple hydrogel group was treated with lipopolysaccharide and blank hydrogel. The drug- loaded hydrogel group was treated with lipopolysaccharide and drug-loaded hydrogel. The inhibitor group was treated with lippolysaccharide, drug-loaded hydrogel, and PI3K inhibitor LY294002. 24 hours later, mRNA expression of inflammation-related factors was detected by qRT-PCR. Western blot assay was utilized to detect the protein expression of inflammation-related factors and PI3K/AKT signaling pathway. (3) MC3T3-E1 cells were inoculated in four groups. The blank group was cultured without any material. The simple hydrogel group was treated with blank hydrogel. The drug-loaded hydrogel group was treated with drug-loaded hydrogel. The inhibitor group was treated with drug-loaded hydrogel and PI3K inhibitor LY294002 for 7 days. Alkaline phosphatase staining was performed. mRNA expression levels of osteogenic factors were detected by qRT-PCR. The protein expression levels of the PI3K/AKT signaling pathway were detected by western blot assay. RESULTS AND CONCLUSION: (1) The results of CCK-8 assay and live/dead cell staining showed that the two kinds of hydrogels had no cytotoxic effect and had good cytocompatibility. (2) Compared with the blank group, the mRNA and protein expression levels of interleukin 6, tumor necrosis factor α, and interleukin 1β were significantly increased (P < 0.05), and the protein expression levels of p-AKT, p-PI3K, p-p65, and p-IκBα were significantly increased in the lipopolysaccharide group (P < 0.05). Compared with lipopolysaccharide group, mRNA and protein expression levels of the above indexes were decreased in drug-loaded hydrogel group (P < 0.05). Compared with the drug-loaded hydrogel group, the mRNA and protein expression levels of the above indexes were decreased in the inhibitor group (P < 0.05). (3) The activity of alkaline phosphatase in drug-loaded hydrogel group was higher than that in the blank group, simple hydrogel group, and inhibitor group (P < 0.05). Compared with blank group, the mRNA expression levels of alkaline phosphatase, Runx2, osteocalcin, and type I collagen were increased (P < 0.05), and the protein expression levels of p-AKT and p-PI3K were increased in the simple hydrogel group (P < 0.05). Compared with the simple hydrogel group, the mRNA and protein expression levels of the above indexes were increased in the drug-loaded hydrogel group (P < 0.05). Compared with the drug-loaded hydrogel group, the mRNA and protein expression levels of the above indexes were decreased in the inhibitor group (P < 0.05). (4) These findings conclude that chitosan/sodium glycerophosphate/sodium alginate/leonurine hydrogel has anti-inflammatory and osteogenic effects, which may be related to the regulation of PI3K/AKT signaling pathway. [ABSTRACT FROM AUTHOR]

Leonurine is an alkaloid unique to the Leonurus genus, which has many biological activities, such as uterine contraction, anti-inflammation, anti-oxidation, regulation of cell apoptosis, anti-tumor, angiogenesis, anti-platelet aggregation, and inhibition of vasoconstriction. This paper summarizes the extraction methods, synthetic pathways, biosynthetic mechanisms, pharmacokinetic properties, pharmacological effects in various diseases, toxicology, and clinical trials of leonurine. To facilitate a successful transition into clinical application, intensified efforts are required in several key areas: structural modifications of leonurine to optimize its properties, comprehensive pharmacokinetic assessments to understand its behavior within the body, thorough mechanistic studies to elucidate how it works at the molecular level, rigorous safety evaluations and toxicological investigations to ensure patient wellbeing, and meticulously conducted clinical trials to validate its efficacy and safety profile. [ABSTRACT FROM AUTHOR]

This research explores the potential of leonurine, a component found in motherwort, to inhibit intestinal inflammatory response in patients of inflammatory bowel disease (IBD) through dietary means with a particular emphasis on how it influences the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway. Using colon epithelial cell lines, HT29 and Caco-2, we set up models to study the impact of inflammation, oxidative stress, and apoptosis induced by lipopolysaccharide (LPS). In addition, we employed Nrf2 knockout cell lines to understand its function in a better way. The introduction of leonurine is aimed to evaluate its ability to reduce cell damage and inflammatory reactions by activating the Nrf2–heme oxygenase-1 (HO-1) pathway. Results indicated that LPS exposure led to inflammation, oxidative stress, apoptosis, and potential ferroptosis. It was observed that Nrf2 acted as a suppressor in this scenario; its absence increased cell susceptibility to these challenges. Treatment with leonurine notably decreased inflammation, oxidative stress, and apoptosis; however, these protective effects were significantly reduced in cells lacking Nrf2. This confirms that leonurine can inhibit oxidative stress, inflammatory response, apoptosis, and potential ferroptosis mechanism of colon epithelial cells by activating Nrf2. This study not only enhances our understanding of how leonurine regulates IBD but also highlights the importance of Nrf2 by shedding light on IBD development and potential nutritional approaches for its management. [ABSTRACT FROM AUTHOR]

Endometritis is a common disease in postpartum cows, characterized by delayed uterine recovery due to endometrial inflammation. Although antibiotics and hormones are commonly used, they have certain limitations. One potential alternative is using motherwort extract, specifically leonurine, which exhibits anti-inflammatory properties. However, leonurine's exact molecular mechanism of action remains unclear. In this study, 40 mice were randomly divided into four groups: a control group, endometritis model group, LPS + leonurine group (30 mg/kg), and LPS + dexamethasone group (5 mg/kg). Transcriptomic analysis revealed that leonurine modulates multiple signaling pathways, including JAK-STAT/PI3K-Akt, and influences the expression of key genes, such as Prlr, Socs2, Col1a1, and Akt1. Furthermore, leonurine effectively reduces levels of inflammatory cytokines, such as tumor necrosis factor-alpha (TNF-α), interleukin (IL)-6, and IL-1β (p < 0.01), which play a crucial role in regulating acute endometritis. Additionally, leonurine helps maintain cholesterol homeostasis and attenuates inflammation through the peroxisome proliferator-activated receptor (PPAR) signaling pathway by modulating genes such as Cyp27a1, Hmgcs1, and Scd2. These findings suggest that leonurine has a protective effect against LPS-induced endometritis and that its anti-inflammatory properties involve multiple pathways and targets, which are potentially mediated by regulating signaling pathways such as JAK-STAT/PI3K-Akt and PPAR. [ABSTRACT FROM AUTHOR]

Leonurine (SCM-198) was discovered as one of the active constituents of the Herba Leonuri (HL). Now it can be artificially synthesized. Several recent researches has proven that it exhibits antiinflammatory effect in several systems in animal models and cell culture in vitro. The key mechanism involves downgrading the activity of nuclear transcription factor- κB (NF- κB), thereby inhibiting the phosphorylation of several signal pathways such as PI3K/Akt, MAPK, ERK, and JNK, or upregulating the activity of Nrf2 related pathways, resulting in downregulated expression of inflammatory cytokines such as tumor necrosis factor- α (TNF- α), IL-1β, IL-2, IL-6, IL-8, inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), chemokines, adhesion molecules, etc. Owing to the advantages of high safety and efficiency, the ease of administration, as well as its effectiveness in many organs and systems, leonurine has a widely prospect for future research and clinical applications. This article reviews the progress in the fundamental research of leonurine in multiple inflammation-related disease, and it could be expect to offer new possibilities for the treatment of these disease. [ABSTRACT FROM AUTHOR]

Objective To investigate the effects of leonurine on autophagy and apoptosis of chondrocytes and the role of PTEN-induced putative kinase 1 (PINK1)/Parkinson's disease protein (Park) signaling pathway in osteoarthritis (OA) rats. Methods Standard anterior cruciate ligament transection (ACLT) was applied to construct an OA rat model. The successfully modeled rats were divided into OA group, leonurine L group, leonurine M group, and leonurine H group, with 12 rats in each group. Other 12 rats were collected as the sham operation group. HE staining was applied to detect the morphological changes of chondrocytes of rats in each group. Transmission electron microscopy was applied to observe the ultrastructure of chondrocytes of rats in each group. TUNEL method was applied to detect chondrocyte apoptosis of rats in each group. RT-qPCR was applied to detect the mRNA expression levels of PINK1, Parkin, P62, and LC3B in the cartilage tissue of rats in each group. Western blotting was applied to detect the expression levels of PINK1, Parkin, P62, and LC3B proteins in the cartilage tissue of rats in each group. Results Compared to that in the sham surgery group, the cartilage morphology of rats in the OA group was abnormal, the structure was blurry, the cracks appeared on the surface, the number of mitochondria in chondrocytes reduced, and a small amount of autophagosomes were present. The apoptosis rate of chondrocytes and the levels of P62 mRNA and protein in the cartilage tissue increased significantly. The mRNA and protein levels of PINK1, Parkin, and LC3B in the cartilage tissue reduced significantly (P<0.05). Compared to those in the OA group, the cartilage morphology of the rats in the leonurine L, M, H groups gradually recovered, the structure gradually became clear, surface cracks decreased, the number of chondrocytes restored, mitochondria increased, and the number of autophagosomes increased. The apoptosis rate of chondrocytes and the mRNA and protein levels of P62 in the cartilage tissue reduced significantly. The mRNA and protein levels of PINK1, Parkin, and LC3B in the cartilage tissue increased with a dose-dependent manner (P<0.05). Conclusion Leonurine may promote autophagy and inhibit chondrocyte apoptosis in OA rats by activating the PINK1/Parkin signaling pathway. [ABSTRACT FROM AUTHOR]

Leonurine (LEO), an alkaloid isolated from Leonurus spp., has anti‐oxidant, anti‐inflammatory and anti‐apoptotic effects and can prevent damage caused by reactive oxygen species (ROS). These properties suggest that it can improve the maturation rate of oocytes and developmental ability of embryos, which are key parameters in animal breeding. In this study, the effects of LEO on in vitro maturation and early embryonic development in sheep oocytes were evaluated. Among various doses examined (0, 10, 20 and 40 μM), a dose of 20 μM was optimal with respect to the oocyte maturation rate. Compared with estimates in the control group, GSH levels and mitochondrial membrane potential of sheep oocytes treated with 20 μM LEO were significantly higher, and 40 μM LEO would affect oocyte maturation. Additionally, ROS levels were significantly lower, expression levels of the antioxidant genes CAT and SOD1 were significantly higher, and there was no significant difference in GPX3 expression. The Bax/Bcl‐2 ratio and Caspase‐3 expression were significantly reduced in the 20 μM LEO group. During early embryonic development in vitro, the cleavage rate and blastocyst rate were significantly higher in the 20 μM LEO treatment group compared to other groups. GSH levels and mitochondrial membrane potential were significantly higher, while ROS levels were significantly lower, and expression levels of the antioxidant genes CAT, GPX3 and SOD1 were significantly higher in eight‐cell embryos treated with 20 μM LEO than in the control group. The Bax/Bcl‐2 ratio and Caspase‐3 levels were significantly decreased. In summary, LEO can reduce the effect of oxidative stress, improve the oocyte maturation rate and enhance embryonic development. [ABSTRACT FROM AUTHOR]

Leonurine is a natural product unique to the Lamiaceae plant Leonurus japonicus Houtt., and it has attracted attention due to its anti-oxidative stress, anti-apoptosis, anti-fibrosis, and metabolic regulation properties. Also, it plays an important role in the prevention and treatment of nonalcoholic fatty liver disease (NAFLD) through a variety of biological mechanisms, but its mechanism of action remains to be elucidated. Therefore, this study aims to preliminarily explore the mechanisms of action of leonurine in NAFLD. Mice were randomly divided into four groups: the normal control (NC) group, the Model (M) group, the leonurine treatment (LH) group, and the fenofibrate treatment (FB) group. The NAFLD model was induced by a high-fat high-sugar diet (HFHSD) for 12 weeks, and liver pathological changes and biochemical indices were observed after 12 weeks. Transcriptomic analysis results indicated that leonurine intervention reversed the high-fat high-sugar diet-induced changes in lipid metabolism-related genes such as stearoyl-CoA desaturase 1 (Scd1), Spermine Synthase (Sms), AP-1 Transcription Factor Subunit (Fos), Oxysterol Binding Protein Like 5 (Osbpl5), and FK506 binding protein 5 (Fkbp5) in liver tissues. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis results suggest that leonurine may exert its lipid-lowering effects through the AMP-activated protein kinase (AMPK) signaling pathway. Liver lipidomic analysis showed that leonurine could alter the abundance of lipid molecules related to fatty acyl (FAs) and glycerophospholipids (GPs) such as TxB3, carnitine C12-OH, carnitine C18:1-OH, and LPC (20:3/0:0). Molecular biology experiments and molecular docking techniques verified that leonurine might improve hepatic lipid metabolism through the alpha-1A adrenergic receptor (ADRA1a)/AMPK/SCD1 axis. In summary, the present study explored the mechanism by which leonurine ameliorated NAFLD by inhibiting hepatic lipid synthesis via the ADRA1a/AMPK/SCD1 axis.

The Lamiaceae family is renowned for its terpenoid-based medicinal components, but Leonurus , which has traditional medicinal uses, stands out for its alkaloid-rich composition. Leonurine, the principal active compound found in Leonurus , has demonstrated promising effects in reducing blood lipids and treating strokes. However, the biosynthetic pathway of leonurine remains largely unexplored. Here, we present the chromosome-level genome sequence assemblies of Leonurus japonicus , known for its high leonurine production, and Leonurus sibiricus , characterized by very limited leonurine production. By integrating genomics, RNA sequencing, metabolomics, and enzyme activity assay data, we constructed the leonurine biosynthesis pathway and identified the arginine decarboxylase (ADC), uridine diphosphate glucosyltransferase (UGT), and serine carboxypeptidase-like (SCPL) acyltransferase enzymes that catalyze key reactions in this pathway. Further analyses revealed that the UGT–SCPL gene cluster evolved by gene duplication in the ancestor of Leonurus and neofunctionalization of SCPL in L. japonicus , which contributed to the accumulation of leonurine specifically in L. japonicus. Collectively, our comprehensive study illuminates leonurine biosynthesis and its evolution in Leonurus. Leonurine, the principal active compound in Leonurus , has shown significant potential in lowering blood lipids and treating strokes. By employing genome sequencing and multiomics techniques, this study elucidates the genetic basis of leonurine biosynthesis and uncovers the evolutionary history of this potent alkaloid across Lamiales. The findings offer valuable insights into leonurine biosynthesis and its evolutionary trajectory. [ABSTRACT FROM AUTHOR]

Background: Diabetic nephropathy (DN) is a common microvascular complication of diabetes mellitus (DM). Ferroptosis is an atypical form of iron-dependent, modulated cell death that has been shown to occur in human umbilical vein endothelial cells (HUVECs). Leonurine (LEO) is a single active ingredient extracted from Leonurus japonicus Houtt. It has various biological activities, including anti-inflammatory and anti-cancer effects. However, whether LEO affects ferroptosis in DN has yet to be investigated. Methods: An animal model of DN was established by subjecting C57/BL6 mice to a high-fat diet (HFD) while being induced with Streptozotocin (STZ). A cellular model of DN was established by exposing HUVECs to a high glucose (HG) concentration of 30 mM. Results: LEO was found to improve DN and to attenuate the degree of glomerulosclerosis and tubular atrophy in the mouse model. Additionally, it markedly decreased the levels of ferroptosis markers. Molecular analyses revealed that LEO inhibited HG-induced oxidative stress in HUVECs, thereby decreasing endothelial cell (EC) dysfunction. Furthermore, LEO was found to reduce ferroptosis and reverse EC dysfunction by increasing the expression of glutathione peroxidase 4 (GPX4) and nuclear factor erythroid 2-related factor 2 (Nrf2). The suppression of Nrf2 in HG-induced HUVECs inhibited LEO-GPX4 axis-mediated ferroptosis and increased EC dysfunction. Conclusions: LEO exerts anti-DN effects both in vivo and in vitro by suppressing GPX4-mediated EC ferroptosis. Mechanistically, LEO appears to induce Nrf2-mediated GPX4 expression to inhibit ferroptosis, thereby reducing EC dysfunction. This study provides a new perspective on the treatment of diseases using natural medicines. It involves a novel form of cell death that could potentially lead to better treatment of DN.

Yue Xie,1,&ast; Yaning Jin,2,&ast; Shuyue Li,3 Baoxi Shen,4 Liping Ma,5 Lujie Zuo,6 Ya Gao,3 Guofeng Yang3 1Department of Neurology, the Second Medical Center and National Clinical Research Center for Geriatric Disease, Chinese PLA General Hospital, Beijing, 100853, People’s Republic of China; 2Chaoyang Second Retired Cadre Rest Center of Beijing Garrison, Beijing, 100853, People’s Republic of China; 3Department of Geriatrics, the Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, 050000, People’s Republic of China; 4Department of Neurosurgery, the First Medical Centre, Chinese PLA General Hospital, Medical School of Chinese PLA, Beijing, 100853, People’s Republic of China; 5Department of Neurology, Xinzhou People’s Hospital, Shanxi, 034000, People’s Republic of China; 6Department of Otolaryngology, Head and Neck Surgery, Children’s Hospital of Hebei Province, Shijiazhuang, Hebei, 050000, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Ya Gao; Guofeng Yang, Department of Geriatrics, the Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, 050000, People’s Republic of China, Email elegance86@163.com; gf_yanghb@126.comIntroduction: Alzheimer’s disease (AD) is the most common type of dementia, impacting approximately 50 million individuals globally. However, the current treatments available for AD are only symptomatic and have limited efficacy. This study aimed to investigate whether Leonurine could alleviate cognitive dysfunction in a mouse model of AD and explore its underlying molecular mechanisms.Methods: In this study, male APP/PS1 mice were orally administered Leonurine for two consecutive months. The cognitive functions of the mice were then evaluated using novel object recognition (NOR) and Morris water maze (MWM) tests. Hippocampal neuronal damage was observed through Nissl staining, Aβ levels were determined through ELISA, oxidative stress activity was detected through biochemical methods, and the nuclear factor erythroid-2-related factor 2 (Nrf-2) pathway was analyzed using western blot and real-time quantitative polymerase chain reaction analysis.Results: Our results demonstrated that Leonurine treatment markedly improved cognitive functions, as indicated by the improved performance in the model. Additionally, histopathology showed a reduction in hippocampal neuronal damage. This can be attributed to the potential of Leonurine to reduce Aβ 1-40 and Aβ 1-42 levels and alleviate oxidative stress. Its antioxidant effect is linked to the activation of the Nrf-2 signaling pathway in APP/PS1 mice, which promotes Nrf-2 nuclear translocation and expression of HO-1 and NQO-1.Conclusion: These findings suggest that Leonurine could be explored further as it could emerge as a promising drug for AD treatment.Keywords: Alzheimer’s disease, cognitive deficit, Leonurine, Nrf-2, oxidative stress

Leonurine refers to the desiccated aerial portion of a plant in the Labiatae family. The primary bioactive constituent of Leonurine is an alkaloid, Leonurine alkaloid (Leo), renowned for its substantial therapeutic efficacy in the treatment of gynecological disorders, in addition to its broad-spectrum antineoplastic capabilities. Over recent years, the pharmacodynamic mechanisms of Leo have garnered escalating scholarly interest. Leo exhibits its anticancer potential by means of an array of mechanisms, encompassing the inhibition of neoplastic cell proliferation, induction of both apoptosis and autophagy, and the containment of oncogenic cell invasion and migration. The key signal transduction pathways implicated in these processes include the Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand (TRAIL), the Phosphoinositide3-Kinase/Serine/ Threonine Protein Kinase (PI3K/AKT), the Signal Transducer and Activator of Transcription 3 (STAT3), and the Mitogen-Activated Protein/Extracellular SignalRegulated Kinase (MAP/ERK). This paper commences with an exploration of the principal oncogenic cellular behaviors influenced by Leo and the associated signal transduction pathways, thereby scrutinizing the mechanisms of Leo in the antineoplastic sequence of events. The intention is to offer theoretical reinforcement for the elucidation of more profound mechanisms underpinning Leo’s anticancer potential and correlating pharmaceutical development. [ABSTRACT FROM AUTHOR]

Parkinson's disease (PD) is a neurodegenerative disorder associated with α‐synuclein aggregation and dopaminergic neuron loss in the midbrain. There is evidence that psychological stress promotes PD progression by enhancing glucocorticoids‐related oxidative damage, however, the mechanisms involved are unknown. The present study demonstrated that plasma membrane phospholipid peroxides, as determined by phospholipidomics, triggered ferroptosis in dopaminergic neurons, which in turn contributed to stress exacerbated PD‐like motor disorder in mice overexpressing mutant human α‐synuclein. Using hormonomics, we identified that stress stimulated corticosteroid release and promoted 15‐lipoxygenase‐1 (ALOX15)‐mediated phospholipid peroxidation. ALOX15 was upregulated by α‐synuclein overexpression and acted as a fundamental risk factor in the development of chronic stress‐induced parkinsonism and neurodegeneration. Further, we demonstrated the mechanism by which corticosteroids activated the PKC pathway and induced phosphatidylethanolamine‐binding protein‐1 (PEBP1) to form a complex with ALOX15, thereby facilitating ALOX15 to locate on the plasma membrane phospholipids. A natural product isolated from herbs, leonurine, was screened with activities of inhibiting the ALOX15/PEBP1 interaction and thereby attenuating membrane phospholipid peroxidation. Collectively, our findings demonstrate that stress increases the susceptibility of PD by driving membrane lipid peroxidation of dopaminergic neurons and suggest the ALOX15/PEBP1 complex as a potential intervention target. [ABSTRACT FROM AUTHOR]

Purpose: To elucidate the effect of leonurine on the proliferation, invasion, migration, and angiogenetic potential of breast cancer cells. Methods: Human breast cancer cell line (MDA-MB-231) and normal breast cell line, (SK-BR-3) were cultured. Both cell lines were treated with 200, 400, or 800 μM leonurine and cultured for 0 (control), 24, 48, or 72 h. Cell counting kit-8 (CCK8) and colony formation assays were performed to measure cell viability and proliferation. Invasion and migration were evaluated using in vitro invasion and wound healing assays, respectively, while angiogenesis was evaluated by the formation of branching point structures. Furthermore, phosphorylation of PI3K, AKT, and mTOR were assessed by Western blot. Cell viability, invasion, migration, and angiogenesis were further investigated in media including 740Y-P, 800 μM leonurine, and 800 μM leonurine plus 740Y-P. Results: Leonurine inhibited the proliferation of breast cancer cells and weakened breast cancer cell invasion, migration, and angiogenetic potential in a dose-dependent manner (p < 0.05). Furthermore, leonurine repressed PI3K/AKT/mTOR pathway by reducing the phosphorylation of PI3K, AKT, and mTOR. Leonurine also inhibited breast cancer progression (p < 0.05). Conclusion: Leonurine inhibits breast cancer progression by inhibiting PI3K/AKT/mTOR pathway, and is thus, a potential agent for the management of breast cancer. [ABSTRACT FROM AUTHOR]

Ischemic stroke (IS) constitutes the leading cause of global morbidity and mortality. Neuroprotectants are essential to ameliorate the clinical prognosis, but their therapeutic outcomes are tremendously compromised by insufficient delivery to the ischemic lesion and intricate pathogenesis associated with neuronal damage, oxidative stress, inflammation responses, blood-brain barrier (BBB) dysfunction, etc. Herein, a biomimetic nanosystem (Leo@NM-Lipo) composed of neutrophil membrane-fused nanoliposomal leonurine (Leo) is constructed, which can not only efficiently penetrate and repair the disrupted BBB but also robustly remodel the harsh cerebral microenvironment to reverse ischemia-reperfusion (I/R) injury. More specifically, the neutrophil membrane inherits the BBB penetrating, infarct core targeting, inflammation neutralization, and immune evasion properties of neutrophils, while Leo, a naturally occurring neuroprotectant, exerts pleiotropic effects to attenuate brain damage. Remarkably, comprehensive investigations disclose the critical factors influencing the targetability and therapeutic performances of biomimetic nanosystems. Leo@NM-Lipo with a low membrane protein-to-lipid ratio of 1:10 efficiently targets the ischemic lesion and rescues the injured brain by alleviating neuronal apoptosis, oxidative stress, neuroinflammation, and restoring BBB integrity in transient middle cerebral artery occlusion (tMCAO) rats. Taken together, our study provides a neutrophil-mimetic nanoplatform for targeted IS therapy and sheds light on the rational design of biomimetic nanosystems favoring wide medical applications.

Objective: Leonurine is a bioactive alkaloid compound extracted from Leonurus japonicus Houtt, which potentially has immunomodulatory effects. The immunomodulatory effect and mechanism of leonurine on monocyte derived dendritic cells (moDCs) from healthy donors (HDs) and multiple myeloma (MM) patients were investigated for the first time. Methods: Peripheral blood from HDs and MM patients was isolated for peripheral blood mononuclear cells (PBMCs). The generation of moDCs was conducted by the incubation of monocytes from PBMCs in the medium consisting of RPMI 1640 medium, 2 mmol/L L-glutamine, 5% human serum, 800 U/mL GM-CSF, 500 U/mL IL-4, 100 U/mL penicillin and 0.1 mg/mL streptomycin. During the incubation of 7 days, the cells were administrated with 1 µM leonurine or 1 × PBS as the control group. On the 8th day, cells were harvested. The expression of maturation associated surface markers CD40, CD83, and HLA-DR on moDCs was analyzed by flow cytometry. Moreover, moDCs with or without 1 µM leonurine administration were evaluated by LC-MS/MS for metabolomics which was further analyzed for the potential mechanism of leonurine on moDCs. Results: The proportion of moDCs in the harvested cells was significantly higher in the HD group (n = 14) than in the MM patient group (n = 11) (p = 0.000). Leonurine significantly enhanced the median fluorescence intensity of CD83, HLA-DR and CD40 expression on HD-moDCs (n = 14; p = 0.042, p = 0.013, p = 0.084) as well as MMpaitent-moDCs (n = 11; p = 0.020, p = 0.006, p = 0.025). The metabolomics data showed that in moDCs (HD, n = 15), 18 metabolites in the pathway of arachidonic acid metabolism showed significant differences between the leonurine group and the control group (VIP all >1 and P all <0.05). To be specific, 6-Keto-PGE1, 8,9-DHET, 11 (R)-HETE, 12-Keto-LTB4, 12-OxoETE, 15 (S)-HETE, 15-Deoxy-Delta12,14-PGJ2, 15-Keto-PGF2a, 20-COOH-LTB4, Lecithin, PGA2, PGB2, PGE2, PGF2a, PGG2, Prostacyclin were significantly upregulated in the leonurine group than in the control group, while Arachidonic Acid and TXB2 were significantly downregulated in the leonurine group than in the control group. Conclusion: Leonurine significantly promotes the maturation of moDCs derived from HDs and MM patients, the mechanism of which is related to arachidonic acid metabolism. [ABSTRACT FROM AUTHOR]

Atherosclerosis (AS) is a chronic inflammatory disease with high morbidity and mortality worldwide and is the pathologic basis of cerebral-cardiovascular disease. Cerebral infarction is caused by cerebral ischemia in the deep brain stem or cerebral hemisphere, and small vessel atherosclerosis is the pathophysiological basis of lacunar cerebral infarction. In this study, we found that inhibition of PARP1 in neuronal cells increased the proliferation rate of cells, reduced the concentration of cholesterol in neuronal cells, inhibited the expression and secretion of proinflammatory factors and promoted the expression and secretion of proangiogenic factors. In addition, the reverse transport process of cholesterol and autophagy process were activated in neuron cells by detecting the target proteins using western blotting analysis, and further experiments found that this process might be mediated by activation of the PI3K/AKT/mTOR signalling pathway. Thus, we thought inhibition of PARP1 might be a new therapeutic method for cerebral-cardiovascular disease.

Atherosclerosis (AS) is the most prevalent cardiovascular disease and remains the major contributor to death and mortality globally. Leonurine (LEO) is a unique alkaloid compound with protective effects on the cardiovascular system. However, the exact mechanisms underlying its cardiovascular-protecting action are still not fully elucidated. The methyltransferase 3 (METTL3), the catalytic core of the N6-methyladenosine modification (m6A) methyltransferase complex, has been shown to inhibit autophagy and exacerbate the process of AS via regulation of m6A modification of mRNA. We aimed to determine whether the inhibited effect of LEO on AS is related to METTL3-mediated AKT1S1 stability. The apolipoprotein E (ApoE) knockout mice was subjected to a high-fat diet (HFD), and THP-1 derived macrophages was exposed to oxidized low-density lipoprotein (ox-LDL), to establish the animal and cellular models of AS, respectively. We found that LEO effectively improved AS and reduced the plaque area and inflammation via diminishing macrophage lipid accumulation and remodeling the lipid metabolism profile. LEO activated ox-LDL-induced macrophage autophagy, enhancing lipid metabolism decrease, according to the lipidomic and molecular biology analyses. Additionally, LEO caused a marked increase in autophagy marker levels in mouse models with advanced AS. Furthermore, we found that LEO reactivated autophagy and reversed lipid accumulation by suppressing METTL3 expression. The m6A-seq from ox-LDL-induced macrophages showed that a total of five autophagy-related mRNA transcripts (AKT1S1, AKT1, RB1CC1, CFLAR , and MTMR4) were altered, and AKT1S1 was significantly upregulated by LEO. Mechanistically, LEO-mediated regulation of METTL3 decreased AKT1S1 expression by attenuating its mRNA stability. Silencing AKT1S1 inhibited LEO-METTL3 axis-mediated autophagy and enhanced lipid accumulation in ox-LDL-induced macrophages. The study first revealed that LEO exerts anti-atherosclerotic effect by activating METTL3-mediated macrophage autophagy in vivo and in vitro. The mechanism of LEO was further found to be the enhancement of METTL3-mediated AKT1S1 stability to activate autophagy thereby reducing lipid accumulation. This study provides a new perspective of natural medicines on the treatment of AS via an epigenetic manner. [Display omitted] [ABSTRACT FROM AUTHOR]

Prostate cancer is a leading cause of cancer-associated death in males. Leonurine (Leo) is a pleiotropic anti-tumor agent isolated from traditional Chinese herb that was used in gynecologic treatments. However, its pharmacological effect against prostate cancer progression remains unclear. Here, we showed that Leo dose dependently inhibited prostate cancer cell proliferation, promoted cell apoptosis, and induced cell cycle arrest. Moreover, we noticed that miR-18a-5p was downregulated and the solute carrier family 40 member 1 (SLC40A1) is upregulated by Leo treatment. SLC40A1 knockdown by siRNA abrogated the inhibitory effect of Leo on prostate cancer progression. Notably, Leo also significantly inhibited prostate cancer progression in a subcutaneous xenograft tumor mouse model in vivo. This study further unveiled the mechanism by which Leo inhibited prostate cancer progression, which provides a promising potential for its future clinical application.

Abstract Background Pre‐eclampsia (PE) is mainly attributed to the inflammation of trophoblast cells in pregnant women, which results in damage to the maternal organs and growth retardation of the fetus. Alkaloid leonurine (LNR) is a plant compound and has anti‐inflammatory effects. Here we aimed to investigate the effects of LNR on human and mouse trophoblast cells and the underlying mechanisms. Methods The levels of the inflammatory factors in trophoblast cells under lipopolysaccharides (LPS) stimulation were analyzed with ELISA. Western blot was employed to examine the protein expression. Trophoblast cells in Mammalian ste20‐like kinase 1 (MST1−/−) or wild type (WT) mice were isolated to examine the expression of signal molecules in the nuclear factor‐κB (NF‐κB) pathway. Concentration‐dependent activity of NF‐κB was examined. The regulation of LNR and MST1 in MST1−/− trophoblast cells was studied as well. Results Our data showed that LNR exhibited anti‐inflammatory effects and suppressed the NF‐κB signaling by inhibiting LPS‐induced inflammation in trophoblast cells. LNR upregulated the expression of MST1, and the anti‐inflammatory role of LNR was greatly relieved in MST1‐knockout trophoblast cells, although it displayed weak roles in NF‐κB signaling. Conclusion LNR exhibits anti‐inflammatory effects on human and mouse trophoblast cells by upregulating MST1 in the NF‐κB signal pathway.

Prostate cancer is a leading cause of cancer-associated death in males. Leonurine (Leo) is a pleiotropic anti-tumor agent isolated from traditional Chinese herb that was used in gynecologic treatments. However, its pharmacological effect against prostate cancer progression remains unclear. Here, we showed that Leo dose dependently inhibited prostate cancer cell proliferation, promoted cell apoptosis, and induced cell cycle arrest. Moreover, we noticed that miR-18a-5p was downregulated and the solute carrier family 40 member 1 (SLC40A1) is upregulated by Leo treatment. SLC40A1 knockdown by siRNA abrogated the inhibitory effect of Leo on prostate cancer progression. Notably, Leo also significantly inhibited prostate cancer progression in a subcutaneous xenograft tumor mouse model in vivo. This study further unveiled the mechanism by which Leo inhibited prostate cancer progression, which provides a promising potential for its future clinical application. [ABSTRACT FROM AUTHOR]

Leonurine has been shown to have excellent anti-myocardial ischemia effects. Our previous studies suggested that cardiac protection by leonurine during myocardial ischemia appeared to be inextricably linked to its regulation of the liver. At present, however, there are few mechanistic studies of leonurine and its regulation of hepatic metabolism against ischemic injury. In this study, a metabolomics approach was developed to give a global view of the metabolic profiles of the heart and liver during myocardial ischemia. Principal component analysis and orthogonal partial least squares discrimination analysis were applied to filter differential metabolites, and a debiased sparse partial correlation analysis was used to analyze the correlation of the differential metabolites between heart and liver. As a result, a total of thirty-one differential metabolites were identified, six in the myocardial tissue and twenty-five in the hepatic tissue, involving multiple metabolic pathways including glycine, serine and threonine, purine, fatty acid, and amino acid metabolic pathways. Correlation analysis revealed a net of these differential metabolites, suggesting an interaction between hepatic and myocardial metabolism. These results suggest that leonurine may reduce myocardial injury during myocardial ischemia by regulating the metabolism of glycine, serine and threonine, purine, fatty acids, and amino acids in the liver and heart. [ABSTRACT FROM AUTHOR]

Leonurine (Leo) has been found to have neuroprotective effects against cerebral ischemic injury. However, the exact molecular mechanism underlying its neuroprotective ability remains unclear. The aim of the present study was to investigate whether Leo could provide protection through the nitric oxide (NO)/nitric oxide synthase (NOS) pathway. We firstly explored the effects of NO/NOS signaling on oxidative stress and apoptosis in in vivo and in vitro models of cerebral ischemia. Further, we evaluated the protective effects of Leo against oxygen and glucose deprivation (OGD)-induced oxidative stress and apoptosis in PC12 cells. We found that the rats showed anxiety-like behavior, and the morphology and number of neurons were changed in a model of photochemically induced cerebral ischemia. Both in vivo and in vitro results show that the activity of superoxide dismutase (SOD) and glutathione (GSH) contents were decreased after ischemia, and reactive oxygen species (ROS) and malondialdehyde (MDA) levels were increased, indicating that cerebral ischemia induced oxidative stress and neuronal damage. Moreover, the contents of NO, total NOS, constitutive NOS (cNOS) and inducible NOS (iNOS) were increased after ischemia in rat and PC12 cells. Treatment with L-nitroarginine methyl ester (L-NAME), a nonselective NOS inhibitor, could reverse the change in NO/NOS expression and abolish these detrimental effects of ischemia. Leo treatment decreased ROS and MDA levels and increased the activity of SOD and GSH contents in PC12 cells exposed to OGD. Furthermore, Leo reduced NO/NOS production and cell apoptosis, decreased Bax expression and increased Bcl-2 levels in OGD-treated PC12 cells. All the data suggest that Leo protected against oxidative stress and neuronal apoptosis in cerebral ischemia by inhibiting the NO/NOS system. Our findings indicate that Leo could be a potential agent for the intervention of ischemic stroke and highlighted the NO/NOS-mediated oxidative stress signaling. [ABSTRACT FROM AUTHOR]

Purpose: Cisplatin has been widely accepted as an effective chemotherapy drug with various side effects, including nephrotoxicity. The mechanisms of cisplatin-induced acute kidney injury (AKI) are complex, and there are limited renoprotective approaches. Leonurine is the main active compound of a Chinese herb and has recently been reported to have a protective effect on the kidneys. This study aimed to verify the renoprotective effect of leonurine in attenuating cisplatin-induced AKI and explore the potential associated mechanisms. Methods: C57BL/6 mice were divided into four groups (Sham, Cisplatin, Leonurine, and Cisplatin + Leonurine). Mice in the leonurine-treated groups were pretreated with a daily intraperitoneal injection of 25 mg/kg leonurine. AKI was induced by injecting cisplatin once intraperitoneally at 20 mg/kg body weight. Mice were killed on day 5. Kidney injury was assessed using a serum biochemical and histological assay. Apoptosis was evaluated using a terminal deoxyribonucleotide transferase-mediated dUTP nick-end labeling (TUNEL) staining assay and Western blot. Antioxidant enzymes were detected using commercial kits. The improvement in inflammasome activation, mitochondrial dysfunction, and endoplasmic reticulum stress (ERS) were assessed by polymerase chain reaction (PCR) and Western blot, respectively. Results: Leonurine treatment improved kidney function by preventing renal tubular injury and apoptosis. Expression of nucleotide-binding leucine-rich repeat and pyrin domain containing protein 3 (NLRP3) inflammasome components and inflammatory cytokines, mitochondrial dysfunction, and ERS were all alleviated by leonurine. Conclusion: The results indicate that leonurine plays a protective role in cisplatin-induced AKI and may represent an effective multi-targeted intervention strategy. [ABSTRACT FROM AUTHOR]

Purpose: Motherwort (Leonurus cardiaca), a medicinal plant of the Lamiaceae family, has a great diversity and wide distribution in Iran. It is essential due to having leonurine with blood dilution and muscle relaxation effects. Research method: To evaluate the response of the Iranian Motherwort ecotypes in drought environments, an experiment was carried out in split plot design based on RCBD with three replications. Three irrigation levels were considered as main plots, including %100, one-third, and two-third of field capacity, and four ecotypes as sub-plots including Kerman, Taleghan, Sarab and Khansar. The experiment was conducted at Horticultural Research Station, the University of Tehran, in Karaj. Findings: The results showed that water stress affected morphological traits significantly. Water stress had a significant increasing effect on the phenolic and flavonoid contents and antioxidant activity of Motherwort ecotypes. The highest and the lowest total phenol content were observed in Taleghan and Kerman ecotypes, 7.34 and 4.21 mg/mg fresh weight, respectively. The amount and the percentage of leonurine were increased by water stress. The highest and the lowest values of leonurine were observed in the ecotypes of Taleghan and Sarab, respectively, in all three levels of irrigation. There were significant correlations between dry weight (-0.43), antioxidant (0.36), proline (-0.35), catalase (0.4), and the percentage of leonurine. Research limitations: There were no limitations to the report. Originality/Value: It was found a significant variation among the ecotypes in response to water stress. The ecotypes of Taleghan and Sarab were the most tolerant and the most susceptible ecotypes to drought stress among the studied ecotypes.

Natural products, those molecules derived from nature, have been used by humans for thousands of years to treat ailments and diseases. More recently, these compounds have inspired chemists to use natural products as structural templates in the development of new drug molecules. One such compound is leonurine, a molecule isolated and characterized in the tissues of Herb leonuri. This molecule has received attention from scientists in recent years due to its potent anti-oxidant, anti-apoptotic, and anti-inflammatory properties. More recently researchers have shown leonurine to be useful in the treatment of cardiovascular and nervous system diseases. Like other natural products such as paclitaxel and artemisinin, the historical development of leonurine as a therapeutic is very interesting. Therefore, this review provided an overview of natural product discovery, through to the development of a potential new drug. Content will summarize known plant sources, the pathway used in the synthesis of leonurine, and descriptions of leonurine’s pharmacological properties in mammalian systems.

Background and purpose: Evidence indicates that ferroptosis plays a key role in acute kidney injury induced by cisplatin. The Nrf2/NRF2 pathway regulates oxidative stress, lipid peroxidation and positively regulates cisplatin‐induced acute kidney injury, but its effect along with the alkaloid leonurine, found in motherwort, on ferroptosis after such acute kidney injury remains unclear. Experimental Approach: The anti‐ferroptotic effects of Nrf2 and leonurine were assessed in a mouse model of cisplatin‐induced acute kidney injury. In vitro, the effects of leonurine on erastin‐ and RSL3‐induced HK‐2 human PTEC ferroptosis were examined. Key Results: Nrf2 deletion induced ferroptosis‐related protein expression and iron accumulation in vivo, aggravating cisplatin‐induced acute kidney injury. Leonurine activated Nrf2 and prevented iron accumulation, lipid peroxidation and ferroptosis in vitro, being abolished in siNrf2‐treated cells. Moreover, leonurine potently inhibited cisplatin‐induced renal damage, as assessed by of serum creatinine, blood urea nitrogen, kidney injury molecule‐1 and NGAL. Importantly, leonurine activated the Nrf2 antioxidative pathway and preventing changes in ferroptosis‐related morphological and biochemical indicators, malondialdehyde level, SOD and GSH depletion, and GPX4 and xCT down‐regulation, in cisplatin‐induced acute kidney injury. Nrf2 KO mice were more susceptible to ferroptosis after cisplatin‐induced acute kidney injury than control mice. The protective effects of leonurine on acute kidney injury and ferroptosis were largely abolished in Nrf2 KO mice. Conclusion and Implications: These data suggest that renal protective effects of Nrf2 activation on cisplatin‐induced acute kidney injury are achieved, at least partially, by inhibiting lipid peroxide‐mediated ferroptosis, highlighting the potential of leonurine in acute kidney injury treatment. [ABSTRACT FROM AUTHOR]

Osteoporosis bears an imbalance between bone formation and resorption, which is strongly related to oxidative stress. The function of leonurine on bone marrow-derived mesenchymal stem cells (BMSCs) under oxidative stress is still unclear. Therefore, this study was aimed at identifying the protective effect of leonurine on H2O2 stimulated rat BMSCs. We found that leonurine can alleviate cell apoptosis and promote the differentiation ability of rat BMSCs induced by oxidative stress at an appropriate concentration at 10 μM. Meanwhile, the intracellular ROS level and the level of the COX2 and NOX4 mRNA decreased after leonurine treatment in vitro. The ATP level and mitochondrial membrane potential were upregulated after leonurine treatment. The protein level of PINK1 and Parkin showed the same trend. The mitophage in rat BMSCs blocked by 3-MA was partially rescued by leonurine. Bioinformatics analysis and leonurine-protein coupling provides a strong direct combination between leonurine and the PI3K protein at the position of Asp841, Glu880, Val882. In conclusion, leonurine protects the proliferation and differentiation of BMSCs from oxidative stress by activating mitophagy, which depends on the PI3K/Akt/mTOR pathway. The results showed that leonurine may have potential usage in osteoporosis and bone defect repair in osteoporosis patients. [ABSTRACT FROM AUTHOR]

BackgroundLeonurus japonicus Houtt has an obvious efficacy on cardiovascular diseases. As the most representative component in the herb, leonurine has attracted increasing attention for its potential in myocardial ischemia. However, its protective mechanism against myocardial ischemia remains incompletely elucidated.ObjectivesThe present study aimed to reveal the potential mechanism of leonurine in acute myocardial ischemia using a strategy combining metabolomics and network pharmacology.MethodsFirst, a metabolomics method was proposed to identify the differential metabolites of plasma in rats. Then, network pharmacology was performed to screen candidate targets of leonurine against acute myocardial ischemia. A compound-reaction-enzyme-gene network was thus constructed with the differential metabolites and targets. Finally, molecular docking was carried out to predict the binding capability of leonurine with key targets.ResultsA total of 32 differential metabolites were identified in rat plasma, and 16 hub genes were detected through network pharmacology. According to the results of compound-reaction-enzyme-gene network and molecular docking, what was screened included six key targets (GSR, CYP2C9, BCHE, GSTP1, TGM2, and PLA2G2A) and seven differential metabolites (glycerylphosphorylcholine, lysophosphatidylcholine, choline phosphate, linoleic acid, 13-HpODE, tryptophan and glutamate) with four important metabolic pathways involved: glycerophospholopid metabolism, linoleic acid metabolism, tryptophan metabolism and glutamate metabolism. Among them, glycerophospholipid and tryptophan metabolism were shown to be important, since the regulation of leonurine on these two pathways was also observed in our previous metabolomics study conducted on clinical hyperlipidemia patients.ConclusionThis is the first study of its kind to reveal the underlying mechanism of leonurine against acute myocardial ischemia through a strategy combining metabolomics and network pharmacology, which provides a valuable reference for the research on its future application.

• A fluorescent probe, leonurine-P, was facilely prepared from coupling FITC and the small-molecule phytodrug leonurine. • Leonurine-P can be used to visualize spatiotemporal dynamics processes such as drug uptake and distribution in C. elegans and living cells. • Leonurine-P can be further explored in standard fluorescence imaging or flow cytometric platforms to allow quantitative analyses. Caenorhabditis elegans (C. elegans) has been recognized for being a useful model organism in small-molecule drug screens and drug efficacy investigation. However, there remain bottlenecks in evaluating such processes as drug uptake and distribution due to a lack of appropriate chemical tools. This study aims to prepare fluorescence-labeled leonurine as an example to monitor drug uptake and distribution of small molecule in C. elegans and living cells. FITC-conjugated leonurine (leonurine-P) was synthesized and characterized by LC/MS, NMR, UV absorption and fluorescence intensity. Leonurine-P was used to stain C. elegans and various mammalian cell lines. Different concentrations of leonurine were tested in conjunction with a competing parent molecule to determine whether leonurine-P and leonurine shared the same biological targets. Drug distribution was analyzed by imaging. Fluorometry in microplates and flow cytometry were performed for quantitative measurements of drug uptake. The UV absorption peak of leonurine-P was 490∼495 nm and emission peak was 520 nm. Leonurine-P specifically bound to endogenous protein targets in C. elegans and mammalian cells, which was competitively blocked by leonurine. The highest enrichment levels of leonurine-P were observed around 72 h following exposure in C. elegans. Leonurine-P can be used in a variety of cells to observe drug distribution dynamics. Flow cytometry of stained cells can be facilely carried out to quantitatively detect probe signals. The strategy of fluorescein-labeled drugs reported herein allows quantification of drug enrichment and visualization of drug distribution, thus illustrates a convenient approach to study phytodrugs in pharmacological contexts. [Display omitted] [ABSTRACT FROM AUTHOR]

Oxidative stress and inflammation play important roles in pleurisy. Leonurine (Leo) has been confirmed to exert antioxidative and antiinflammatory effects in many preclinical experiments, but these effects have not been studied in pleurisy. The aim of this study was to explore the therapeutic effect and mechanism of Leo in a carrageenan (CAR)‐induced pleurisy model. In this study, we found that the increase of reactive oxygen species (ROS), myeloperoxidase (MPO), and malondialdehyde (MDA) and decrease of glutathione (GSH) induced by CAR could be reversed by the treatment of Leo. Leo effectively reduced the levels of proinflammatory cytokines interleukin‐1β (IL‐1β), tumor necrosis factor‐α (TNF‐α), and the percentages of mature macrophages and increased the levels of antiinflammatory cytokines (IL‐10). Furthermore, Western blotting revealed that Leo significantly activated the Nrf2 pathway to restrain the thioredoxin‐interacting protein/NOD‐like receptor protein 3 (TXNIP/NLRP3) and nuclear factor kappa‐B (NF‐κB) pathways. However, the protective effect of Leo was significantly weakened in Nrf2‐deficient mice. These results indicate that Leo confers potent protection against CAR‐induced pleurisy by inhibiting the TXNIP/NLRP3 and NF‐κB pathways dependent on Nrf2, which may serve as a promising agent for attenuating pleurisy. [ABSTRACT FROM AUTHOR]

The quality parameters of Leonurus sibiricus L. aerial part crude drugs were evaluated. Fifteen crude drugs were collected from various locations throughout Thailand. The transverse section of the stem of L. sibiricus showed quadrangular character highlighted the ribs with angular collenchyma. The epidermis was uniseriate with abundant glandular trichomes distribution. Prismatic calcium oxalate prisms were found in the stem medullary parenchyma. The histological character of crude drug powder showed bordered pitted vessel, fragment of fiber, glandular trichome, prism crystal, spiral vessel, starch granule, and stomata. The loss on drying, total ash, acid-insoluble ash, and moisture contents should be not more than 8.18, 15.28, 4.04, and 8.91 g/100 g dry weight, whereas ethanol and water-soluble extractive values should be not less than 7.67, and 17.21 g/100 g of dry weight, respectively. Leonurine in the crude drugs were analyzed by capillary electrophoresis (CE) with photodiode array detector. The ethanolic extraction performed by Soxhlet apparatus yielded 18.86 ± 4.09 g/100 g dry weight. The electropherogram detected at 277 nm showed the migration time of leonurine at 6.2 min. The developed CE was found to be valid for leonurine quantification in L. sibiricus ethanolic extract. The contents of leonurine in 15 crude drugs ranged from 0.79 to 4.23 mg/g with the average of 2.38 ± 1.10 mg/g dry weight. This study established the pharmacognostic specification of L. sibiricus crude drug in Thailand with special reference to a bioactive compound, leonurine. CE was beneficial technique for the analysis of leonurine in L. sibiricus aerial parts.

Inflammatory bowel disease (IBD) is a common chronic inflammatory gastrointestinal disease. The therapeutic strategies of IBD are limited. IBD mouse models were established by administering 4% dextran sodium sulfate (DSS), which were further treated with Leonurine (7.5, 15, 30 mg/kg). The disease phenotypes, cell apoptosis, inflammation factors and oxidative stress related chemicals were evaluated. In addition, the potential related mechanism was also explored. Consequently, Leonurine ameliorated IBD-associated disease phenotypes and increase colon lengths and inhibited intestinal cell apoptosis in DSS-induced IBD mice. In addition, Leonurine reduced the expression of inflammation factors and oxidative stress level in DSS-induced IBD mice. Finally, Leonurine inhibited TLR4/NF-κB signaling pathway and activated of Nrf2/HO-1 signaling pathway. Leonurine can ameliorate IBD-induced apoptosis, inflammation response and oxidative stress via the activation of Nrf2/HO-1 signaling pathway and suppression of TLR4/NF-κB pathway. [ABSTRACT FROM AUTHOR]

Min Lin, Chunyu Pan, Wenbin Xu, Jingwei Li, Xueqiong Zhu Department of Obstetrics and Gynecology, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325027, People’s Republic of ChinaCorrespondence: Xueqiong ZhuDepartment of Obstetrics and Gynecology, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325027, People’s Republic of ChinaTel +86 577 8800 2796Fax +86 577 88002796Email zjwzzxq@163.comBackground: Cisplatin-based neoadjuvant chemotherapy and concurrent radiotherapy and chemotherapy are the main treatment for advanced cervical cancer. However, the development of multidrug resistance (MDR) leads to chemotherapy failure, tumor recurrence and poor survival. In this research, we investigated the effect and corresponding mechanism of leonurine on cisplatin sensitivity of cervical cancer cells.Methods: Anti-cervical cancer efficacy of leonurine and leonurine combined with cisplatin was examined in C33A and Ms751 cells. The cell counting kit-8 assay and bromodeoxyuridine assay were applied for measuring cell proliferation. CompuSyn software was used to calculate the combination index and assess the synergistic effect of leonurine and cisplatin on cell proliferation. The cell cycle distribution and cell apoptosis were analyzed by flow cytometry. The expression of cleaved caspase-3, poly ADP-ribose polymerase (PARP), B-cell lymphoma-2 associated X (BAX), B-cell lymphoma-2 (BCL-2), P glycoprotein (P-Gp) protein and multiple drug resistance protein 1 (MRP1) was analyzed by Western blotting.Results: Leonurine had time- and dose-dependent anti-proliferative effects on C33A and MS751 cells. Leonurine and cisplatin combination was more efficacious in inhibiting the growth of cervical cancer cells than either of the two drugs. The combined application has shown that the cervical cancer cells were arrested at G1 phase after treatments. Moreover, flow cytometry analysis indicated that the combined treatment could cause more cell apoptosis than the single drug treatment. Consistently, combined treatment elevated BAX/BCL-2 ratio, and the expression of BAX, PARP and cleaved caspase-3 proteins. Mechanistic investigations uncovered that the tumor-inhibiting effects of the co-treatment were mediated by repressing MDR, including MRP1 and P-Gp protein, thereby enhancing the efficiency of cisplatin.Conclusion: Leonurine and cisplatin have synergistic antitumorigenic effects on cervical cancer. Combination with leonurine may serve as a novel strategy for enhancing cisplatin sensitivity via the inhibition of the expression of MRP1 and P-Gp.Keywords: cervical cancer, leonurine, cisplatin, multidrug resistance

Background: Diabetic nephropathy (DN) is a common microvascular complication of diabetes mellitus (DM). Ferroptosis is an atypical form of iron-dependent, modulated cell death that has been shown to occur in human umbilical vein endothelial cells (HUVECs). Leonurine (LEO) is a single active ingredient extracted from Leonurus japonicus Houtt . It has various biological activities, including anti-inflammatory and anti-cancer effects. However, whether LEO affects ferroptosis in DN has yet to be investigated., Methods: An animal model of DN was established by subjecting C57/BL6 mice to a high-fat diet (HFD) while being induced with Streptozotocin (STZ). A cellular model of DN was established by exposing HUVECs to a high glucose (HG) concentration of 30 mM., Results: LEO was found to improve DN and to attenuate the degree of glomerulosclerosis and tubular atrophy in the mouse model. Additionally, it markedly decreased the levels of ferroptosis markers. Molecular analyses revealed that LEO inhibited HG-induced oxidative stress in HUVECs, thereby decreasing endothelial cell (EC) dysfunction. Furthermore, LEO was found to reduce ferroptosis and reverse EC dysfunction by increasing the expression of glutathione peroxidase 4 (GPX4) and nuclear factor erythroid 2-related factor 2 (Nrf2). The suppression of Nrf2 in HG-induced HUVECs inhibited LEO-GPX4 axis-mediated ferroptosis and increased EC dysfunction., Conclusions: LEO exerts anti-DN effects both in vivo and in vitro by suppressing GPX4-mediated EC ferroptosis. Mechanistically, LEO appears to induce Nrf2-mediated GPX4 expression to inhibit ferroptosis, thereby reducing EC dysfunction. This study provides a new perspective on the treatment of diseases using natural medicines. It involves a novel form of cell death that could potentially lead to better treatment of DN., Competing Interests: The authors declare no conflict of interest., (© 2024 The Author(s). Published by IMR Press.)

Leonurine is an alkaloid unique to the Leonurus genus, which has many biological activities, such as uterine contraction, anti-inflammation, anti-oxidation, regulation of cell apoptosis, anti-tumor, angiogenesis, anti-platelet aggregation, and inhibition of vasoconstriction. This paper summarizes the extraction methods, synthetic pathways, biosynthetic mechanisms, pharmacokinetic properties, pharmacological effects in various diseases, toxicology, and clinical trials of leonurine. To facilitate a successful transition into clinical application, intensified efforts are required in several key areas: structural modifications of leonurine to optimize its properties, comprehensive pharmacokinetic assessments to understand its behavior within the body, thorough mechanistic studies to elucidate how it works at the molecular level, rigorous safety evaluations and toxicological investigations to ensure patient wellbeing, and meticulously conducted clinical trials to validate its efficacy and safety profile., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Liu, Sun, Tang, Peng and Peng.)

Thrombosis is a general pathological phenomenon during severe disturbances to homeostasis, which plays an essential role in cardiovascular and cerebrovascular diseases. Leonurine (LEO), isolated from Leonurus japonicus Houtt, showes a crucial role in anticoagulation and vasodilatation. However, the properties and therapeutic mechanisms of this effect have not yet been systematically elucidated. Therefore, the antithrombotic effect of LEO was investigated in this study. Hematoxylin-Eosin staining was used to detect the thrombosis of zebrafish tail. Fluorescence probe was used to detect the reactive oxygen species. The biochemical indexes related to oxidative stress (lactate dehydrogenase, malondialdehyde, superoxide dismutase and glutathione) and vasodilator factor (endothelin-1 and nitric oxide) were analyzed by specific commercial assay kits. Besides, we detected the expression of related genes (fga, fgb, fgg, pkcα, pkcβ, vwf, f2) and proteins (PI3K, phospho-PI3K, Akt, phospho-Akt, ERK, phospho-ERK FIB) related to the anticoagulation and fibrinolytic system by quantitative reverse transcription and western blot. Beyond that, metabolomic analyses were carried out to identify the expressions of metabolites associated with the anti-thrombosis mechanism of LEO. Our in vivo experimental results showed that LEO could improve the oxidative stress injury, abnormal platelet aggregation and coagulation dysfunction induced by adrenalin hydrochloride. Moreover, LEO restored the modulation of amino acids and inositol metabolites which are reported to alleviate the thrombus formation. Collectively, LEO attenuates adrenalin hydrochloride-induced thrombosis partly via modulating oxidative stress, coagulation cascade and platelet activation and amino acid and inositol metabolites. [ABSTRACT FROM AUTHOR]

Influenza A virus (H1N1), a swine-origin influenza A virus, causes seasonal epidemics that result in severe illnesses and deaths. Leonurine has been reported to function as an anti-inflammatory agent with protective effects on nervous, urinary and cardiovascular systems. However, the therapeutic effects of leonurine on the pneumonia caused by H1N1 infection remain unclear. Hematoxylin and eosin staining was performed to evaluate the lung injuries of mice infected by H1N1. The amount of immune cells was analyzed by flow cytometry. Enzyme-linked immunosorbent assay was used to evaluate the alteration of multiple cytokines in lung tissues. Real-time quantitative polymerase chain reaction assay was performed to investigate the ribonucleic acid (RNA) levels of certain genes. The protein levels in toll-like receptor 4/nuclear factor kappa-light-chain-enhancer of activated B cells (TLR4/NF-κB) signaling were estimated by western blot assay. Leonurine treatment significantly inhibited the mortality caused by H1N1 infection. Leonurine treatment (60 mg/kg) alleviated the lung injuries caused by virus infection. The inflammatory cell accumulation and cytokine expression were inhibited by the leonurine administration. Leonurine inhibited the mRNA expression of pro-inflammatory cytokines in the lung homogenates at day 5 postinfection. Leonurine regulated the TLR4/NF-κB signaling in the lung homogenates of H1N1-infected mice at day 5 postinfection. Leonurine protects against H1N1 infection-induced pneumonia in mice. [ABSTRACT FROM AUTHOR]

The present study was conducted to explore the anti-acute myeloid leukaemia (AML) effects of leonurine. HL-60 and U-937 cells were used to assess the antileukaemia effect of leonurine in vitro, and HL-60 and U-937 xenograft nude mice were used to evaluate its antitumour effect in vivo. Leonurine inhibited the proliferation of HL-60 and U-937 cells in a time- and dose-dependent manner. Moreover, leonurine therapy prevented the growth of tumours in both xenograft animal models. Leonurine could induce apoptosis in HL-60 and U-937 cells. The cytotoxic effects of le- onurine on HL-60 and U-937 cells were associated with an increased ratio of Bax/Bcl-2, activation of caspase-3, caspase-8 and caspase-9, and increased expression of cytochrome c in the cytoplasm. Leonurine inhibited activation of the PI3K/Akt pathway in HL-60 and U-937 cells by lowering the phosphorylation levels of PI3K and Akt. Our results indicate that leonurine is a potential anti-AML agent, and this activity may be associated with its repression of the phosphorylation of PI3K and Akt. [ABSTRACT FROM AUTHOR]

Thrombosis is a general pathological phenomenon during severe disturbances to homeostasis, which plays an essential role in cardiovascular and cerebrovascular diseases. Leonurine (LEO), isolated from Leonurus japonicus Houtt, showes a crucial role in anticoagulation and vasodilatation. However, the properties and therapeutic mechanisms of this effect have not yet been systematically elucidated. Therefore, the antithrombotic effect of LEO was investigated in this study. Hematoxylin-Eosin staining was used to detect the thrombosis of zebrafish tail. Fluorescence probe was used to detect the reactive oxygen species. The biochemical indexes related to oxidative stress (lactate dehydrogenase, malondialdehyde, superoxide dismutase and glutathione) and vasodilator factor (endothelin-1 and nitric oxide) were analyzed by specific commercial assay kits. Besides, we detected the expression of related genes (fga, fgb, fgg, pkcα, pkcβ, vwf, f2) and proteins (PI3K, phospho-PI3K, Akt, phospho-Akt, ERK, phospho-ERK FIB) related to the anticoagulation and fibrinolytic system by quantitative reverse transcription and western blot. Beyond that, metabolomic analyses were carried out to identify the expressions of metabolites associated with the anti-thrombosis mechanism of LEO. Our in vivo experimental results showed that LEO could improve the oxidative stress injury, abnormal platelet aggregation and coagulation dysfunction induced by adrenalin hydrochloride. Moreover, LEO restored the modulation of amino acids and inositol metabolites which are reported to alleviate the thrombus formation. Collectively, LEO attenuates adrenalin hydrochloride-induced thrombosis partly via modulating oxidative stress, coagulation cascade and platelet activation and amino acid and inositol metabolites.

Leonurine has been shown to have excellent anti-myocardial ischemia effects. Our previous studies suggested that cardiac protection by leonurine during myocardial ischemia appeared to be inextricably linked to its regulation of the liver. At present, however, there are few mechanistic studies of leonurine and its regulation of hepatic metabolism against ischemic injury. In this study, a metabolomics approach was developed to give a global view of the metabolic profiles of the heart and liver during myocardial ischemia. Principal component analysis and orthogonal partial least squares discrimination analysis were applied to filter differential metabolites, and a debiased sparse partial correlation analysis was used to analyze the correlation of the differential metabolites between heart and liver. As a result, a total of thirty-one differential metabolites were identified, six in the myocardial tissue and twenty-five in the hepatic tissue, involving multiple metabolic pathways including glycine, serine and threonine, purine, fatty acid, and amino acid metabolic pathways. Correlation analysis revealed a net of these differential metabolites, suggesting an interaction between hepatic and myocardial metabolism. These results suggest that leonurine may reduce myocardial injury during myocardial ischemia by regulating the metabolism of glycine, serine and threonine, purine, fatty acids, and amino acids in the liver and heart.

Leonurine (Leo) has been found to have neuroprotective effects against cerebral ischemic injury. However, the exact molecular mechanism underlying its neuroprotective ability remains unclear. The aim of the present study was to investigate whether Leo could provide protection through the nitric oxide (NO)/nitric oxide synthase (NOS) pathway. We firstly explored the effects of NO/NOS signaling on oxidative stress and apoptosis in in vivo and in vitro models of cerebral ischemia. Further, we evaluated the protective effects of Leo against oxygen and glucose deprivation (OGD)-induced oxidative stress and apoptosis in PC12 cells. We found that the rats showed anxiety-like behavior, and the morphology and number of neurons were changed in a model of photochemically induced cerebral ischemia. Both in vivo and in vitro results show that the activity of superoxide dismutase (SOD) and glutathione (GSH) contents were decreased after ischemia, and reactive oxygen species (ROS) and malondialdehyde (MDA) levels were increased, indicating that cerebral ischemia induced oxidative stress and neuronal damage. Moreover, the contents of NO, total NOS, constitutive NOS (cNOS) and inducible NOS (iNOS) were increased after ischemia in rat and PC12 cells. Treatment with L-nitroarginine methyl ester (L-NAME), a nonselective NOS inhibitor, could reverse the change in NO/NOS expression and abolish these detrimental effects of ischemia. Leo treatment decreased ROS and MDA levels and increased the activity of SOD and GSH contents in PC12 cells exposed to OGD. Furthermore, Leo reduced NO/NOS production and cell apoptosis, decreased Bax expression and increased Bcl-2 levels in OGD-treated PC12 cells. All the data suggest that Leo protected against oxidative stress and neuronal apoptosis in cerebral ischemia by inhibiting the NO/NOS system. Our findings indicate that Leo could be a potential agent for the intervention of ischemic stroke and highlighted the NO/NOS-mediated oxidative stress signaling.

Introduction: The key transformed T cell transcription factor recombination activation gene protein 2 (RAG2) is regulated during inflammation to allow for the acquisition of effector T cells functions. The present study was designed to investigate whether stress signals elicited by leonurine (LEO) could lead to the degradation of RAG2 through v-akt murine thymoma viral oncogene homolog (AKT) signaling in lung fibrosis. Methods: A total of 120 female mice were randomly divided into five groups (Group I–V): Normal group, bleomycin (BLM), BLM+LEO 50 mg/kg/d, BLM+LEO 100 mg/kg/d, and BLM+LEO 50 mg/kg/d+LY294002. Hematoxylin-eosin, Masson's, and terminal deoxynucleotidyl transferase dUTP nick-end labeling staining were performed to observe the pathomorphological changes. The expression of CD3+, TGF-β, RAG2, and Bcl proteins was examined by immunodetection, while that of E-cadherin (ECAD), AKT, TGF-β1, alpha-actin-2, Bax, and RAG2 was detected by Western blot analysis. Results: The level of T lymphocytes was reduced sharply in LEO-treated mice as compared to the other groups. The AKT signal was greatly inhibited in the BLM group and activated with LEO treatment on day 14. In addition, RAG2 was attenuated by LEO on day 14 and day 28. LY294002 could reverse the expression of AKT and RAG2 on day 28. Remarkably, the low dose of LEO has a greater protective efficacy as compared to the high-dose LEO group in terms of pulmonary fibrosis, T cell inactivation, and apoptosis in alveolar cells. Conclusion: The results of the present study suggested that LEO has a protective effect on lung fibrosis with possible mechanisms of attenuating apoptosis and inflammation via the upregulation of the AKT signal in transformed T cells by suppressing the expression and activity of RAG2. [ABSTRACT FROM AUTHOR]

The quality parameters of Leonurus sibiricus L. aerial part crude drugs were evaluated. Fifteen crude drugs were collected from various locations throughout Thailand. The transverse section of the stem of L. sibiricus showed quadrangular character highlighted the ribs with angular collenchyma. The epidermis was uniseriate with abundant glandular trichomes distribution. Prismatic calcium oxalate prisms were found in the stem medullary parenchyma. The histological character of crude drug powder showed bordered pitted vessel, fragment of fiber, glandular trichome, prism crystal, spiral vessel, starch granule, and stomata. The loss on drying, total ash, acid-insoluble ash, and moisture contents should be not more than 8.18, 15.28, 4.04, and 8.91 g/100 g dry weight, whereas ethanol and water-soluble extractive values should be not less than 7.67, and 17.21 g/100 g of dry weight, respectively. Leonurine in the crude drugs were analyzed by capillary electrophoresis (CE) with photodiode array detector. The ethanolic extraction performed by Soxhlet apparatus yielded 18.86 ± 4.09 g/100 g dry weight. The electropherogram detected at 277 nm showed the migration time of leonurine at 6.2 min. The developed CE was found to be valid for leonurine quantification in L. sibiricus ethanolic extract. The contents of leonurine in 15 crude drugs ranged from 0.79 to 4.23 mg/g with the average of 2.38 ± 1.10 mg/g dry weight. This study established the pharmacognostic specification of L. sibiricus crude drug in Thailand with special reference to a bioactive compound, leonurine. CE was beneficial technique for the analysis of leonurine in L. sibiricus aerial parts. [ABSTRACT FROM AUTHOR]

Acute kidney injury (AKI) is a common and serious global health problem with high risks of mortality and the development of chronic kidney diseases. Leonurine is a unique bioactive component from Leonurus japonicus Houtt. and exerts antioxidant, antiapoptotic or anti-inflammatory properties. This study aimed to explore the benefits of leonurine on AKI and the possible mechanisms involved, with a particular foc on the regulation of ferroptosis and endoplasmic reticulum (ER) stress. Our results showed that leonurine exhibited prominent protective effects against AKI, as evidenced by the amelioration of histopathological alterations and reduction of renal dysfunction. In addition, leonurine significantly suppressed ferroptosis in AKI both in vivo and in vitro by effectively restoring ultrastructural abnormalities in mitochondria, decreasing ASCL4 and 4-HNE levels, scavenging reactive oxygen species (ROS), as well as increasing GPX4 and GSH levels. In parallel, leonurine also markedly mitigated ER stress via down-regulating PERK, eIF-2α, ATF4, CHOP and CHAC1. Further studies suggested that ER stress was closely involved in erastin-induced ferroptosis, and leonurine protected tubular epithelial cells in vitro by inhibiting ER stress-associated ferroptosis via regulating ATF4/CHOP/ASCL4 signalling pathway. Mechanistically, ATF4 silencing in vitro regulated CHOP and ACSL4 expressions, ultimately weakening both ER stress and ferroptosis. Notably, analyses of single-cell RNA sequencing data revealed that ATF4, CHOP and ASCL4 in renal tubular cells were all abnormally upregulated in patients with AKI compared to healthy controls, suggesting their contributions to the pathogenesis of AKI. Altogether, these findings suggest that leonurine alleviates AKI by inhibiting ER stress-associated ferroptosis via regulating ATF4/CHOP/ASCL4 signalling pathway, thus providing novel mechanisms for AKI treatment. • Ferroptosis and endoplasmic reticulum (ER) stress are implicated in acute kidney injury (AKI). • Leonurine alleviates AKI by significantly suppressing ferroptosis and mitigating ER stress. • ER stress is closely involved in the occurrence of ferroptosis of renal tubular epithelial cells. • Leonurine inhibits ER stress-associated ferroptosis via regulating ATF4/CHOP/ASCL4 pathway. • Analyses of scRNA-seq data reveal that ATF4, CHOP and ASCL4 in renal tubules are upregulated in patients with AKI. [ABSTRACT FROM AUTHOR]

• Leo alleviated VCM-induced renal injury and pathological damage, and decreased oxidative stress in kidneys and HK-2 cells. • Leo restored mitochondrial numbers in VCM nephrotoxicity kidneys and reduced mitochondrial ROS levels in HK-2 cells. • Leo activated the PPARγ protein expression in VCM nephrotoxicity kidney tissues. • Leo inhibited the TLR4/NF-κB/TNF-α inflammation pathway in VCM nephrotoxicity kidneys. Vancomycin (VCM) is the first-line antibiotic for severe infections, but nephrotoxicity limits its use. Leonurine (Leo) has shown protective effects against kidney damage. However, the effect and mechanism of Leo on VCM nephrotoxicity remain unclear. In this study, mice and HK-2 cells exposed to VCM were treated with Leo. Biochemical and pathological analysis and fluorescence probe methods were performed to examine the role of Leo in VCM nephrotoxicity. Immunohistochemistry, q-PCR, western blot, FACS, and Autodock software were used to verify the mechanism. The present results indicate that Leo significantly alleviates VCM-induced renal injury, morphological damage, and oxidative stress. Increased intracellular and mitochondrial ROS in HK-2 cells and decreased mitochondrial numbers in mouse renal tubular epithelial cells were reversed in Leo-administrated groups. In addition, molecular docking analysis using Autodock software revealed that Leo binds to the PPARγ protein with high affinity. Mechanistic exploration indicated that Leo inhibited VCM nephrotoxicity via activating PPARγ and inhibiting the TLR4/NF-κB/TNF-α inflammation pathway. Taken together, our results indicate that the PPARγ inhibition and inflammation reactions were implicated in the VCM nephrotoxicity and provide a promising therapeutic strategy for renal injury. [ABSTRACT FROM AUTHOR]

The present study was designed to investigate the protective effects of leonurine, a compound purified from Herba Leonuri that is active on ischemic rat behavior and cortical neurons, and explore the underlying mechanism. The general rat activity, cortical neuron morphology, superoxide dismutase (SOD), malondialdehyde (MDA), g-aminobutyric acid (GABA) and glutamate decarboxylase 67 (GAD67) levels were measured. We found leonurine significantly improve the general activity of rats in an open-field test, which was associated with attenuated neuronal damage induced by ischemia. Moreover, serum SOD activity was significantly greater, MDA level lower in the leonurine group as compared with ischemia group. In addition, GABA content in the cerebral cortex was significantly greater in high-dose leonurine group. Correspondingly, GAD67 protein level coincided with the GABA level. Taken together, our results demonstrated that leonurine attenuated brain injury during ischemia via antioxidative and anti-excitotoxicity effects by targeting GABA and leonurine might become a useful adjuvant neuroprotective agent.

Background and aims: Non-alcoholic steatohepatitis (NASH) is the hepatic manifestation of metabolic syndrome and is characterized by steatosis, inflammation, and fibrosis. We aim to characterize the hepatoprotective effects of Leonurine hydrochloride (LH) and the possible pathway in a cell and rodent model of diet-induced steatohepatitis (NASH). Methods: For in vitro studies, Palmitic acid (PA) and free fatty acid (FFA) induced HepG2 and HL7702 steatosis cell models were used. For in vivo studies, NASH was induced by feeding mice MCD diet. These mice received either placebo or LH at three different doses (50、100、200 mg/kg/day) for 6 weeks. Histological staining’s, and commercially available kits for ALT and AST and hepatic contents of TG, TC, MDA, SOD, and GSH were used to assess NASH. Furthermore, relative liver protein and gene expression levels were determined by Western Blot and qPCR, respectively. Results: After establishing NASH models, LH treatment improved lipid accumulation, hepatic contents of TG, TC, and expression levels of ALT and AST in dose-dependent manner. Also, LH improved MDA, SOD, and GSH expression levels. The results of RT-PCR and Western blotting showed that LH upregulated the expression of AMPK phosphorylation and downregulated SREBP-1c and its target genes expression level. Conclusions: Our data reveal the promising role of Leonurine hydrochloride in the prevention and treatment of NASH, in vitro and in vivo. This effect may be partially mediated by the AMPK/SREBP1 pathway. These findings provide a novel therapeutic target for the clinical treatment of NASH.