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Over the past few decades, significant research has been conducted on tissue‐engineered constructs for cartilage repair. However, there is a growing interest in addressing subchondral bone repair along with cartilage regeneration. This study focuses on a bilayer tissue engineering scaffold loaded with icariin (ICA) and quercetin (QU) for simultaneous treatment of knee joint cartilage and subchondral bone defects. The cytotoxicity of dual‐layer scaffolds loaded with ICA and QU was assessed through live/dead cell staining. Subsequently, these dual‐layer scaffolds loaded with ICA and QU were implanted into cartilage and subchondral bone defects in Sprague–Dawley (SD) rats. The repair effects were evaluated through macroscopic observation, computed tomography, and immunohistochemistry. After 12 weeks of implantation of dual‐layer scaffolds loaded with ICA and QU into the cartilage and bone defects of SD rats, better repair effects were observed in both cartilage and bone defects compared to the blank control group. We found that the dual‐layer tissue‐engineered scaffold loaded with ICA and QU had excellent biocompatibility and could effectively repair articular cartilage and subchondral bone injuries, showing promising prospects for clinical applications. [ABSTRACT FROM AUTHOR]

Improving the reproductive ability and fertility of male ruminants is a central concern in animal husbandry. Phytogenic feed additives, known for their anti-inflammatory, antioxidant, and immunomodulatory properties, are commonly used in animal feed. Icariin (ICA), the primary flavonoid glucoside derived from Epimedium, is a traditional tonic in Chinese herbal medicine. However, its potential to enhance the reproductive performance of male ruminants remains unclear. In this study, twelve healthy adult male dairy goats were divided into two groups. The goats received oral administration of ICA at doses of 0 (control) and 50 mg/kg body weight daily for a consecutive period of 80 days during the breeding season. The effects of ICA on the reproductive performance was analyzed by histological examinations, semen quality analysis, and ELISA experiments. ELISA results showed a progressive increase in serum levels of GnIH, LH, and testosterone with the prolonged ICA treatment (p < 0.05). However, the serum concentration of GnRH in the ICA group initially increased, followed by a subsequent decrease (p < 0.05). The hypothalamic concentrations of dopamine (DA) and 5-hydroxytryptamine (5-HT) were significantly higher in the ICA group compared to the control group (p < 0.01). The CASA system analyzed sperm kinematics and revealed that ICA increased ejaculate volume, with both total motile and progressive motile sperm gradually increasing over time (p < 0.05). ICA did not affect the body weight of the goats but significantly increased the organ coefficient of the testes (p < 0.01). Additionally, there was an upregulation of hormone receptor expression in testicular tissue and an improvement in the antioxidant capacity of the testes after ICA treatment (p < 0.01). Furthermore, ICA was implicated in testosterone synthesis by modulating the expression of key enzymes associated with steroidogenesis and promoting the differentiation of spermatogonial stem cell to enhance spermatogenesis. In conclusion, our results indicate that icariin, as a phytogenic feed additive incorporated into the diet of ruminants, offers potential benefits in improving the reproductive performance of male dairy goats. [ABSTRACT FROM AUTHOR]

AbstractObjectives: Reports of traditional Chinese medicine (TCM)-related liver injury have increased over recent years; however, identifying susceptibility-related components and biomarkers remains challenging due to the heterogeneous nature of TCM and idiosyncratic drug-induced liver injury (IDILI). Psoraleae Fructus (PF) and Epimedii Folium (EF), commonly found in TCM prescriptions, have been implicated in IDILI, but their constituents and underlying mechanisms are poorly understood.Methods: In this study, we identified bavachin (Bav) and icariin (Ica) as susceptibility components for IDILI in PF and EF using a TNF-α-mediated mouse model. Lipidomics and transcriptomics were used to investigate their related mechanism.Results: Liver biochemistry and histopathology analyses revealed that co-exposure to Bav, Ica, and a non-toxic dose of TNF-α prestimulation induced significant liver injury, while Bav and Ica alone did not. Lipidomics identified seven differentially abundant metabolites in the Bav/Ica/TNF-α group compared to the Ica/TNF-α or Bav/TNF-α groups, mainly enriched in alpha-linolenic acid (ALA), arachidonic acid (AA), and linoleic acid (LA) metabolic pathways. Additionally, transcriptomics revealed 49 differentially expressed genes (DEGs) in the Bav/TNF-α vs Bav/Ica/TNF-α and Ica/TNF-α vs Bav/Ica/TNF-α groups, primarily associated with the PI3K/AKT/mTOR signaling pathway and sphingolipid metabolism. Integrative lipidomics and transcriptomics analyses identified significant positive correlations between five differential metabolites (DMs) – PC (O-16:0_14:1), PG (22:1_20:3), PI (16:0_14:1), PS (18:0_19:2), and TG (17:0_18:2_22:5) – and ten DEGs – Nr0b2, Btbd19, Btg2, Fam222a, Fam83f, Gtse1, Anln, Gja4, Srrm4, and Zfp13.Conslusions: Collectively, these results suggest that alterations in intracellular metabolism and gene expression levels may contribute to the synergistic induction of IDILI by the incompatible pair Bav and Ica in the presence of TNF-α. [ABSTRACT FROM AUTHOR]

Alzheimer's disease (AD) is a leading cause of dementia. The aging brain is particularly vulnerable to various stressors, including increased levels of ceramide. However, the role of ceramide in neuronal cell senescence and AD progression and whether icariin, a natural flavonoid glucoside, could reverse neuronal senescence remain inadequately understood. In this study, we explore the role of ceramide in neuronal senescence and AD, and whether icariin can counteract these effects. We pretreated HT-22 cells with icariin and then induced senescence with ceramide. Various assays were employed to assess cell senescence, such as reactive oxygen species (ROS) production, cell cycle progression, β-galactosidase staining, and expression of senescence-associated proteins. In vivo studies utilized APP/PS1 mice and C57BL/6J mice injected with ceramide to evaluate behavioral changes, histopathological alterations, and senescence-associated protein expression. Transcriptomics, molecular docking, molecular dynamics simulations, and cellular thermal shift assays were employed to verify the interaction between icariin and P53. The specificity of icariin targeting of P53 was further confirmed through rescue experiments utilizing the P53 activator Navtemadlin. Our data demonstrated that ceramide could induce neuronal senescence and AD-related pathologies, which were reversed by icariin. Moreover, molecular studies revealed that icariin directly targeted P53, and its neuroprotective effects were attenuated by P53 activation, providing evidence for the role of P53 in icariin-mediated neuroprotection. Icariin demonstrates a protective effect against ceramide-induced neuronal senescence by inhibiting the P53 pathway. This identifies a novel mechanism of action for icariin, offering a novel therapeutic approach for AD and other age-related neurodegenerative diseases. [Display omitted] • Ceramide could induce neuronal senescence and Alzheimer's disease-related pathologies. • Icariin demonstrates a protective effect against ceramide-induced neuronal senescence. • Icariin mediated neuroprotective effects by directly targeting P53. • Icariin may serve as a promising therapeutic approach for Alzheimer's disease. [ABSTRACT FROM AUTHOR]

Objectives The aim of this study was to determine the single and combined antidiabetic activity and side effects of the retinoid X receptor agonist bexarotene and the thioredoxin-interacting protein inhibitor and peroxisome proliferator-activated receptor γ and AMP-activated protein kinase activator icariin. Methods The rats were grouped as healthy (control), diabetes, diabetes + bexarotene (20 mg/kg), diabetes + icariin (60 mg/kg), diabetes + bexarotene (10 mg/kg) + icariin (30 mg/kg) low-dose combination and diabetes + bexarotene (20 mg/kg) + icariin (60 mg/kg) high-dose combination groups. Key findings Icariin treatment led to a significant reduction in glucose levels compared with the diabetes control group, a remarkable outcome observed 45 days after the initial application. HbA1c levels of the icariin and low-dose combination treatment groups were significantly lower than in the diabetes group. Notably, icariin treatment also significantly elevated HOMA-β levels, which is indicative of improved β-cell function. Icariin significantly decreased glucose levels at 30 and 120 min in the oral glucose tolerance test. Moreover, it ameliorated hepatocyte degeneration, hepatic cord dissociation, congestion, mononuclear cell infiltration in the liver, and degeneration in the pancreas. Conclusions Icariin treatment exhibited robust antidiabetic effects with fewer side effects than other treatment options in this study. In future studies, long-term and varying doses of icariin will contribute to the development of novel antidiabetic drugs. [ABSTRACT FROM AUTHOR]

BACKGROUND: Previous studies have demonstrated that icariin has important roles in promoting bone formation and inhibiting bone resorption, but its effects on osteoporosis-mediated bone pain have not been reported. OBJECTIVE: To investigate the possible mechanism of icariin alleviating bone pain in postmenopausal senile osteoporosis. METHODS: (1) Animal experiment: 200 C57BL/6 mice were randomly divided into 4 groups: sham group (n=50), model group (n=50), icariin treatment group (n=50), and carbonic anhydrase II inhibitor (Brinzolamide) treatment group (n=50). Ovariectomy was performed on C57BL/6 mice to establish a postmenopausal osteoporosis model in all groups except the sham group. The icariin group was given icariin on the second day after modeling, and pain behavior tests (Von Frey, Hot Plate, and Tail Flick tests) were performed every 2 weeks for 20 weeks. After sampling, bone mass was detected by microCT, osteoclast activity was detected by hematoxylin-eosin and tartrate-resistant acid phosphatase staining, and neuronal morphology and related ion channel expression were detected by tissue immunofluorescence staining. (2) Cell experiment: Osteoclast precursor cells derived from mouse bone marrow were extracted and induced to differentiate into osteoclasts using the RANKL/M-CSF system in vitro and supplemented with icariin of different concentrations (1 and 10 μmol/L). Tartrateresistant acid phosphatase staining was used to detect osteoclast differentiation, ghost pen cyclic peptide staining was used to detect osteoclast actin ring, bone plate absorption assay was used to detect osteoclast osteophagy function, pH value of the system was detected by pH meter, and expression of osteoclast differentiation-related proteins was detected by western blot. In addition, mouse dorsal root ganglion-derived nerve cells were extracted and treated with icariin. Cell counting kit-8 was used to detect neuronal activity and CGRP staining was used to detect neuronal morphology. RESULTS AND CONCLUSION: Compared with the model group, the icariin treatment group had higher bone mineral density, fewer tartrate-resistant acid phosphatase-positive osteoclasts in bone tissue, decreased neuronal activity, and decreased neuronal transient receptor potential vanilloid subtype 1 channel and carbonic anhydrase II expression. Behavioral results showed that the icariin treatment group was less sensitive to pain than the model group. Icariin inhibited osteoclast differentiation and bone phagocytosis in vitro. Icariin enhanced the activity of dorsal root ganglion neurons and inhibited the expression of calcitonin gene-related peptide and transient receptor potential vanilloid subtype 1 channels in dorsal root ganglia in a relatively non-toxic pH range. To conclude, icariin alleviates bone pain caused by postmenopausal osteoporosis by regulating the acidic microenvironment through its effect on osteoclasts. [ABSTRACT FROM AUTHOR]

Objective: The limited ability to regenerate axons after spinal cord injury (SCI) is influenced by factors such as astrocyte activation, reactive proliferation, and glial scar formation. The TGF-β/Smad (transforming growth factor-β/mothers against decapentaplegic homolog) pathway, associated with astrocytic scarring, plays a crucial role in recovery post-injury. This study aims to investigate how icariin (ICA) interacts with reactive astrocytes in the treatment of spinal cord injury. Methods: A rat SCI model was constructed, and the recovery of motor function was observed after treatment with ICA.HE staining, LFB staining, immunofluorescence staining, and Western blotting were employed to assess ICA’s ability to inhibit astrocyte proliferation in rats following spinal cord injury by modulating YAP, as well as to evaluate the reparative effects of ICA on the injured spinal cord tissue. Primary astrocytes were isolated and cultured. Immunoprecipitation-Western Blot (IP-WB) ubiquitination and cytoplasmnuclear separation were employed to assess PPM1B ubiquitination and nuclear translocation. Results: The CatWalk XT gait analysis, BBB (Basso, Beattie, and Bresnahan) score, electrophysiological measurements, HE staining, and LFB staining collectively demonstrated that ICA promotes motor function and tissue recovery following spinal cord injury in rats. Immunofluorescence staining and Western Blot analyses revealed that ICA inhibits astrocyte proliferation in rats post-spinal cord injury by suppressing YAP activity. Furthermore, the activation of YAP by XMU-MP-1 was shown to compromise the efficacy of ICA in these rats after spinal cord injury. Additional immunofluorescence staining and Western Blot experiments confirmed that ICA inhibits TGFβ1-induced astrocyte activation through the regulation of YAP. The knockdown of PPM1B (protein phosphatase, Mg2+/ Mn2+-dependent 1B) in astrocytes was found to inhibit TGFβ signaling. Additionally, YAP was shown to regulate PPM1B ubiquitination and nuclear translocation through immunoprecipitation-Western blot analysis, along with the segregation of cytoplasm and nucleus. Conclusion: Icariin promotes functional recovery in rats after spinal cord injury by inhibiting YAP and regulating PPM1B ubiquitination to inhibiting the activation of reactive astrocytes. [ABSTRACT FROM AUTHOR]

Atopic dermatitis (AD) is a chronic inflammatory skin disorder characterized by increased sensitivity to environmental allergens and irritants. Icariin, a natural compound extracted from the herb Epimedium, has been traditionally used for its potential anti-inflammatory and antioxidant properties. This study aimed to investigate the regulatory effects of icariin on AD-like symptoms and to elucidate its underlying mechanisms. The effects of icariin on TNF-α/IFN-γ-induced HaCaT cell injury were assessed using various assays, including cell counting kit-8 for cell viability, flow cytometry for reactive oxygen species (ROS) levels, and colorimetric assays for malondialdehyde (MDA) levels and superoxide dismutase (SOD) activity. In addition, the study performed enzyme-linked immunosorbent assays to assess cytokines (IL-1β, IL-6, and IL-8) and chemokines (MDC, TARC, and RANTES) levels. Flow cytometry was used to quantify apoptotic rate, while a wound-healing assay was conducted to assess cell migration. The expression of WT1 associated protein (WTAP) and serpin family B member 4 (SERPINB4) at the mRNA and protein levels was determined using qRT-PCR and western blotting, respectively. The associations between WTAP and SERPINB4 were analyzed using RNA immunoprecipitation assay and m6A RNA immunoprecipitation assay. Icariin treatment significantly mitigated TNF-α/IFN-γ-induced oxidative stress, inflammatory response, and apoptosis in HaCaT cells, while also reversing the inhibitory effect on cell migration. Icariin reduced the expression of WTAP in TNF-α/IFN-γ-stimulated HaCaT cells. Overexpression of WTAP reversed the effects of icariin in TNF-α/IFN-γ-stimulated HaCaT cells. WTAP silencing inhibited the mRNA stability of SERPINB4 through the m6A modification. SERPINB4 overexpression attenuated the effects of WTAP silencing on oxidative stress, inflammatory response, apoptosis, and migration of TNF-α/IFN-γ-stimulated HaCaT cells. Icariin treatment downregulated SERPINB4 expression by regulating WTAP in TNF-α/IFN-γ-stimulated HaCaT cells. Icariin ameliorated TNF-α/IFN-γ-induced human immortalized epidermal cell injury through the WTAP/SERPINB4 axis, highlighting the potential for targeted interventions in AD pathogenesis. [ABSTRACT FROM AUTHOR]

Yang Zhang,1 Minjie Zhang,2 Mengying Li,2 Maomao Miao,2 Dan Shou,2 Peijian Tong1 1Institute of Orthopaedics and Traumatology, The First Affiliated Hospital of Zhejiang Chinese Medical University (Zhejiang Provincial Hospital of Chinese Medicine), Hangzhou, 310053, People’s Republic of China; 2School of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Hangzhou, 310053, People’s Republic of ChinaCorrespondence: Dan Shou; Peijian Tong, Email shoudanok@163.com; peijiantong@zcmu.edu.cnBackground: Infected bone defects pose a challenging clinical issue due to an imbalance of osteoclasts (OC) and osteoblasts (OB). Exosomes are crucial for intercellular signaling of OC and OB in bone repair. Icariin, has been shown to regulate the balance between OC and OB. However, the specific mechanisms by which icariin influences exosomes derived from osteoclasts, and subsequently impacts osteoblast activity, remain unclear. This study aims to investigate the effects of icariin-treated osteoclast-derived exosomes (ICA-OC-Exo) on osteoblast function and bone repair in cases of infected bone defects.Methods: We investigated the exosome profile and localization of multivesicular bodies (MVB) and quantification of intraluminal vesicles (ILVs) in osteoclasts by using transmission electron microscopy. Additionally, the expressions of Rab27A and MITF, which are associated with exosome release, were determined through immunofluorescence staining and Western blot. The profiling of exosomal miRNA expression was conducted via miRNA-sequencing. The effects of ICA-OC-Exo on osteoblast differentiation were determined using RT-qPCR, Western blot, alkaline phosphatase staining. Additionally, ICA-OC-Exo was administered into the localized bone defect of the infected bone rat models, and bone formation was assessed using Micro-CT.Results: Icariin increased the presence of MVBs in the cytoplasm through modulation of the MITF/Rab27A signaling pathway, resulting in higher number of ICA-OC-Exo compared to OC-Exo. Additionally, miR-331-3p expression in ICA-OC-Exo was found to be elevated compared to OC-Exo. ICA-OC-Exo was observed to stimulate osteoblast function by targeting FGF23, reducing DKK1, and subsequently upregulating ALP. In the in vivo study, ICA-OC-Exo exhibited the capacity to enhance bone healing at the site of a local bone defect following anti-infection treatment.Conclusion: Icariin enhanced the quantification of OC-Exo and the expression of miRNA-331-3p in OC-Exo, leading to the regulation of osteoblast function via activation of the miRNA-331-3p/FGF23/DKK1 pathway. ICA-OC-Exo demonstrated potential clinical applicability in bone repair of infected bone defects. Keywords: infected bone defects, icariin, osteoclast, exosomes, osteoblast

BACKGROUND: Human periodontal stem cells have a certain inhibitory effect on the pro-inflammatory function of M1-type macrophages, and it is not clear whether icariin, which has anti-inflammatory and other pharmacological activities, can enhance the inhibitory effect of human periodontal stem cells on M1- type macrophages. OBJECTIVE: To investigate the effect of icariin on M1 macrophages after pretreatment of human periodontal stem cells. METHODS: Primary human periodontal stem cells were isolated, cultured and characterized. THP-1 was induced and M1-type macrophages were identified by immunofluorescence staining and PCR. Human periodontal stem cells were cultured with α-MEM complete medium containing concentrations of 10-7, 10-6, 10-5, and 10-4 mol/L icariin, and the cytotoxicity of Icariin on human periodontal stem cells was detected by the CCK-8 assay at 1, 3, 5, and 7 days, respectively. α-MEM complete medium, untreated α-MEM conditioned medium for human periodontal stem cells and α-MEM conditioned medium for human periodontal stem cells pretreated with icariin for 24 hours were conditioned with RPMI-1640 complete medium in a 1:1 ratio for M1-type macrophages in the control, untreated, and pretreated groups, and 24 hours later, the mRNA expression of inflammatory factors in M1 macrophages was detected by RT-PCR. The protein expression of inflammatory factors in M1 macrophages was detected by ELISA. The expression of surface markers and nuclear factor-κB pathway-related proteins in M1/ M2 macrophages was detected by western blot assay. RESULTS AND CONCLUSION: CCK-8 assay results showed that 10-7, 10-6, 10-5, 10-4 mol/L icariin was not cytotoxic to the human periodontal stem cells, and from day 5 onwards, all the concentrations increased the cell viability, and promoted the cell proliferation. 10-4 mol/L icariin was selected for follow-up experiment. RT-PCR and ELISA results showed that compared with the control group, the untreated group and the pretreated group both decreased the expression and secretion of interleukin-1β, interleukin-6, and tumor necrosis factor-α of M1-type macrophages (P < 0.05), and the pretreated group was lower than the untreated group (P < 0.05). Western blot assay results showed that compared with the untreated group, the expression of CD86 was significantly lower in the pretreated group (P < 0.05); compared with the control group, the expression of CD206, a surface marker of M2-type macrophages, was elevated in both the untreated and pretreated groups (P < 0.01), and it was significantly higher in the pretreated group than in the untreated group (P < 0.01). In M1- type macrophages after 24 hours of conditioned culture, compared with the control group, the expression of nuclear factor-κB/P65 was decreased in the untreated group and the pretreated group (P < 0.01), and the expression of p-IκBα was decreased only in the pretreated group (P < 0.01); the expression of both nuclear factor-κB/P65 and p-IκBα was significantly reduced in the pretreated group compared with the untreated group (P < 0.05), while the difference of IκBα in the three groups was not statistically significant. These results indicated that icariin enhanced the inhibitory effect of human periodontal stem cells on M1-type macrophages, and this effect may be related to the inhibition of the nuclear factor-κB signaling pathway of macrophages. [ABSTRACT FROM AUTHOR]

Highlights SPI1 and SMAD5 are upregulated during MC3T3-E1 cell osteogenic differentiation. SPI1 enhances SMAD5 transcription. Icariin increases SMAD5 expression by upregulating SPI1. The SPI1/SMAD5 axis underlies the promoting effect of icariin on MC3T3-E1 cell osteogenic differentiation.

Abstract Cisplatin is a widely used chemotherapeutic drug that can induce ovarian damage. Icariin (ICA), a natural antioxidant derived from Epimedium brevicornum Maxim., has been found to protect against organ injury. The aim of the present study was to investigate whether ICA can exert an ovarian-protective effect on cisplatin induced premature ovarian failure (POF) and the underlying mechanism involved. The preventive effect of ICA was evaluated using body weight, the oestrous cycle, ovarian histological analysis, and follicle counting. ICA treatment increased body weight, ovarian weight, and the number of follicles and improved the oestrous cycle in POF mice. ICA reduced cisplatin-induced oxidative damage and upregulated the protein expression levels of Nrf2, GPX4 and HO-1. Moreover, ICA reduced the expression levels of Bax and γH2AX and inhibited ovarian apoptosis. In addition, ICA activated the Nrf2 pathway in vitro and reversed changes in the viability of cisplatin-induced KGN cells, reactive oxygen species (ROS) levels, lipid peroxidation, and apoptosis, and these effects were abrogated when Nrf2 was knocked down or inhibited. Molecular docking confirmed that ICA promotes the release of Nrf2 by competing with Nrf2 for binding to Keap1. The inhibitory effects of ICA on cisplatin-induced oxidative stress, ferroptosis, and apoptosis may be mediated by its modulatory effects on the Nrf2 pathway, providing a novel perspective on the potential mechanisms by which ICA prevents POF.

Abstract Rheumatoid arthritis (RA) is a chronic autoimmune disease marked by synovitis and cartilage destruction. The active compound, icariin (ICA), derived from the herb Epimedium, exhibits potent anti-inflammatory properties. However, its clinical utility is limited by its water insolubility, poor permeability, and low bioavailability. To address these challenges, we developed a multifunctional drug delivery system—adipose-derived stem cells-exosomes (ADSCs-EXO)-ICA to target active macrophages in synovial tissue and modulate macrophage polarization from M1 to M2. High-performance liquid chromatography analysis confirmed a 92.4 ± 0.008% loading efficiency for ADSCs-EXO-ICA. In vitro studies utilizing cellular immunofluorescence (IF) and flow cytometry demonstrated significant inhibition of M1 macrophage proliferation by ADSCs-EXO-ICA. Enzyme-linked immunosorbent assay, cellular transcriptomics, and real-time quantitative PCR indicated that ADSCs-EXO-ICA promotes an M1-to-M2 phenotypic transition by reducing glycolysis through the inhibition of the ERK/HIF-1α/GLUT1 pathway. In vivo, ADSCs-EXO-ICA effectively accumulated in the joints. Pharmacodynamic assessments revealed that ADSCs-EXO-ICA decreased cytokine levels and mitigated arthritis symptoms in collagen-induced arthritis (CIA) rats. Histological analysis and micro computed tomography confirmed that ADSCs-EXO-ICA markedly ameliorated synovitis and preserved cartilage. Further in vivo studies indicated that ADSCs-EXO-ICA suppresses arthritis by promoting an M1-to-M2 switch and suppressing glycolysis. Western blotting supported the therapeutic efficacy of ADSCs-EXO-ICA in RA, confirming its role in modulating macrophage function through energy metabolism regulation. Thus, this study not only introduces a drug delivery system that significantly enhances the anti-RA efficacy of ADSCs-EXO-ICA but also elucidates its mechanism of action in macrophage function inhibition. Graphical abstract

Icariin (ICA), one of the main active components of Herba Epimedii, is a natural prenylated flavonol glycoside that possesses a wide range of pharmacological effects, including antioxidant, antiosteoporotic, anti-aging, neuroprotective, immunomodulatory, antitumor, and aphrodisiac effects, and prevents numerous health disorders, such as cardiovascular diseases, osteoporosis, cancer, sexual dysfunction, menstrual disorders, neurodegenerative diseases, asthma, chronic inflammation, and diabetes. In the reproductive system, it has been observed that ICA may play a role in preserving fertility by regulating different signalling pathways, such as PI3K/AKT, which improves ovarian function, and ERα/Nrf2, which enhances testicular function and prevents ROS generation. In contrast, regulating the NF/kB signalling pathway causes anti-inflammatory effects, reducing spontaneous abortions. In this study, we review and examine the relevant literature on the therapeutic potential of ICA in reproduction, highlight the various mechanisms of action and limitations for the therapeutic applications of ICA, and summarise and highlight the existing preclinical research on its effects on male and female reproduction. [ABSTRACT FROM AUTHOR]

Retinopathy of prematurity (ROP) is a retinal disease‐causing retinal neovascularization that can lead to blindness. Oxygen‐induced retinopathy (OIR) is a widely used ROP animal model. Icariin (ICA) has anti‐oxidative and anti‐inflammation properties; however, whether ICA has a regulatory effect on OIR remains unclear. In this study, ICA alleviated pathological neovascularization, microglial activation and blood–retina barrier (BRB) damage in vivo. Further results indicated that endothelial cell tube formation, migration and proliferation were restored by ICA treatment in vitro. Proteomic microarrays and molecular mimicry revealed that ICA can directly bind to hexokinase 2 (HK2) and decrease HK2 protein expression in vivo and in vitro. In addition, ICA inhibited the AKT/mTOR/HIF1α pathway activation. The effects of ICA on pathological neovascularization, microglial activation and BRB damage disappeared after HK2 overexpression in vivo. Similarly, the endothelial cell function was revised after HK2 overexpression. HK2 overexpression reversed ICA‐induced AKT/mTOR/HIF1α pathway inhibition in vivo and in vitro. Therefore, ICA prevented pathological angiogenesis in OIR in an HK2‐dependent manner, implicating ICA as a potential therapeutic agent for ROP. [ABSTRACT FROM AUTHOR]

Background. Osteoporosis (OP) is a major problem that increases the mortality and disability rate worldwide. With an increase in the aging population, OP has become a major public threat to human health. Searching for effective and suitable targets for drug treatment in OP has become an urgent need. Objectives. Osteoporosis is a metabolic bone disease characterized by reduced bone mass and density as well as micro-architectural deterioration. Icariin is a flavonoid extracted from plants of the genus Epimedium and has been shown to exert potential anti-OP activity. The present study was designed to observe the effect of icariin on OP and to clarify the underlying mechanisms in ovariectomized (OVX) rats. Materials and methods. Hematoxylin and eosin (H&E) staining, von Kossa staining and micro-computed tomography (micro-CT) confirmed significant bone loss in the OVX group. Protein expression level was detected with western blot analysis. Results. Icariin reversed a trend of increased bone turnover by reducing serum alkaline phosphatase (ALP), procollagen type I N-terminal propeptide (PINP), tartrate-resistant acid phosphatase isoform 5b (TRACP- 5b), and C-telopeptide of type I collagen (CTX-I). Furthermore, icariin decreased sequestosome 1 (p62) and increased microtubule-associated protein 1 light chain 3II/microtubule-associated protein 1 light chain 3I (LC3II/LC3I), autophagy-related protein 7 (Atg7), and Beclin 1 in the femur of OVX rats, improving the indicators of impaired autophagy in OP. Conclusions. Icariin reversed the significant upregulation of the serine/threonine protein kinase (Akt), mammalian target of rapamycin (mTOR) and unc-51-like autophagy activating kinase 1 (ULK1) at Ser757, and the downregulation of p-AMP-activated protein kinase (p-AMPK) and ULK1 phosphorylated at Ser555 in the OVX rats, suggesting that the mechanism of icariin action in OP treatment involves the activation and suppression of the AMPK/ULK1 and AKT/mTOR/ULK1 autophagy pathways, respectively. [ABSTRACT FROM AUTHOR]

BACKGROUND: Studies have shown that chronic apical periodontitis is one of the common inflammatory bone destruction diseases. Icariin can promote osteogenic differentiation, inhibit bone resorption, and may play a protective role in bone destruction caused by chronic apical periodontitis. OBJECTIVE: To investigate the effect of icariin on the proliferation and differentiation of MC3T3-E1 cells in the inflammatory environment stimulated by lipopolysaccharides. METHODS: Lipopolysaccharides were used to stimulate MC3T3-E1 cells to establish an inflammatory environment in vitro, and cell counting kit-8 was used to detect the best concentration and optimal action time of lipopolysaccharides. Cell counting kit-8 was used to detect the optimal concentration of icariin under the stimulation of lipopolysaccharides at a concentration of 1 μg/mL. Alkaline phosphatase detection, Real-time PCR and western blot assay were used to detect the effect of icariin on osteogenic differentiation of MC3T3-E1 cells in the inflammatory environment. Real-time PCR and western blot were used to detect the effects of icariin on the expression of interleukin-1β and interleukin-6 in MC3T3-E1 cells in the lipopolysaccharide-stimulated inflammatory environment RESULTS AND CONCLUSION: Cell counting kit-8 results showed that the optimal concentration of icariin was 0.1 μg/mL. In the inflammatory environment, icariin enhanced the expression of alkaline phosphatase and promoted osteoblast differentiation. Compared with the lipopolysaccharide group, the expression of osteogenesis-related factors alkaline phosphatase and Runx2 was increased in the lipopolysaccharide+icariin group. Compared with the lipopolysaccharide group, the expression levels of inflammation-related factors interleukin-1β and interleukin-6 decreased in the lipopolysaccharide+icariin group. To conclude, lipopolysaccharides weaken the osteogenic ability of MC3T3-E1 cells and aggravate the inflammatory response, but icariin has a protective effect on them. [ABSTRACT FROM AUTHOR]

BACKGROUND: Icariin, with antiinflammatory, antioxygenatory and immunoregulatory effects, can be a potential drug for preventing and treating acute liver injury. OBJECTIVE: To investigate the protective effect and possible mechanism of icariin in mice with acute liver injury induced by carbon tetrachloride. METHODS: Thirty-two Kunming mice were equally and randomly divided into the following groups: normal, model, low-dose icariin and high-dose icariin groups. The low- and high-dose icariin groups were continuously gavaged with icariin (100 and 200 mg/kg, respectively) once a day for 7 continuous days. The normal group and model group were injected with physiological saline (10 mL/kg) at the same time point. After the last administration, all the groups except for the normal group were injected with carbon tetrachloride to induce acute liver injury. The mice were killed 24 hours later, and the liver index was detected. Serum levels of alanine aminotransferase and aspartate aminotransferase were detected by automated biochemical analysis. Tumor necrosis factor α and interleukin 6 levels in serum were detected using ELISA. The levels of superoxide dismutase, glutathione peroxidase and malondialdehyde in liver tissue were detected through a reagent kit. The histopathology changes of the liver were observed by hematoxylin-eosin staining. TUNEL method was used to detect the apoptosis in hepatocytes. Western blot was performed to detect the expression levels of glucose-regulated protein 78 kDa, endoplasmic reticulum stressrelated protein (C/-EBP homologous protein), mixed lineage kinase domain-like protein and Caspase-3 in liver tissue. RESULTS AND CONCLUSION: Compared with the normal group, the liver index and serum levels of alanine aminotransferase, aspartate aminotransferase, tumor necrosis factor α and interleukin 6 were increased in the model group (P < 0.05). Compared with the model group, the above indexes were decreased in the low-dose and high-dose icariin groups (P < 0.05). Compared with the normal group, the activities of superoxide dismutase and glutathione peroxidase in the liver tissue of mice were decreased in the model group (P < 0.05) and the level of malondialdehyde was increased (P < 0.05). Compared with the model group, the activities of superoxide dismutase and glutathione peroxidase were increased in the low- and high-dose icariin groups (P < 0.05) and the level of malondialdehyde was decreased (P < 0.05). Hematoxylin-eosin and TUNEL staining showed that mice in the model group had severe structural destruction of liver tissue, extensive necrosis of hepatocytes and high apoptotic rate of hepatocytes, while the structural destruction of liver tissue and the area of necrosis of hepatocytes in the low- and high-dose icariin groups were significantly milder than those in the model group, and the apoptotic rate of hepatocytes was lower than that in the model group (P < 0.05). Western blot assay showed that the protein expression of glucose-regulated protein 78 kDa, C/-EBP homologous protein, mixed lineage kinase domain-like protein and Caspase-3 in liver tissue of mice in the model group was increased compared with that in the normal group (P < 0.05), while the expression levels of these proteins in liver tissue of mice were significantly reduced after low- and high-dose icariin intervention (P < 0.05). To conclude, icariin can produce a protective effect against carbon tetrachloride-induced acute liver injury, and its mechanism may be related to the regulation of endoplasmic reticulum stress and reduction of programmed necrosis. [ABSTRACT FROM AUTHOR]

New clinical strategies for treating severe bone and cartilage injuries are required, especially for use in combination with implant procedures. For this purpose, p(VCL-co-HEMA) thermosensitive hydrogels have been activated with icariin-loaded nanoparticles to be used as bone-cell-harvesting platforms. Supercritical CO2-SAS technology has been applied to encapsulate icariin, a small molecule that is involved in osteoblastic differentiation. Thus, physical-chemical analysis, including swelling and transmittance, showed the impact of HEMA groups in hydrogel composition. Moreover, icariin (ICA) release from p(VCL-co-HEMA) platforms, including pVCL@ICA nanoparticles, has been studied to evaluate their efficacy in relevant conditions. Finally, the thermosensitive hydrogels' cell compatibility, transplant efficiency, and bone differentiation capacity were tested. This study identifies the optimal formulations for icariin-activated hydrogels for both control and HEMA formulations. Using this technique, osteoblastic sheets that were rich in collagen type I were successfully transplanted and recultivated, maintaining an optimal extracellular matrix (ECM) composition. These findings suggest a new cell-sheet-based therapy for bone regeneration purposes using customized and NP-activated pVCL-based cell platforms. [ABSTRACT FROM AUTHOR]

Highlights: SPI1 and SMAD5 are upregulated during MC3T3-E1 cell osteogenic differentiation. SPI1 enhances SMAD5 transcription. Icariin increases SMAD5 expression by upregulating SPI1. The SPI1/SMAD5 axis underlies the promoting effect of icariin on MC3T3-E1 cell osteogenic differentiation. Background: Dysregulation of osteogenic differentiation is a crucial event during osteoporosis. The bioactive phytochemical icariin has become an anti-osteoporosis candidate. Here, we elucidated the mechanisms underlying the promoting function of icariin in osteogenic differentiation. Methods: Murine pre-osteoblast MC3T3-E1 cells were stimulated with dexamethasone (DEX) to induce osteogenic differentiation, which was evaluated by an Alizarin Red staining assay and ALP activity measurement. The mRNA amounts of SPI1 and SMAD5 were detected by real-time quantitative PCR. Expression analysis of proteins, including osteogenic markers (OPN, OCN and RUNX2) and autophagy-associated proteins (LC3, Beclin-1, and ATG5), was performed by immunoblotting. The binding of SPI1 and the SMAD5 promoter was predicted by the Jaspar2024 algorithm and confirmed by chromatin immunoprecipitation (ChIP) experiments. The regulation of SPI1 in SMAD5 was examined by luciferase assays. Results: During osteogenic differentiation of MC3T3-E1 cells, SPI1 and SMAD5 were upregulated. Functionally, SPI1 overexpression enhanced autophagy and osteogenic differentiation of MC3T3-E1 cells, while SMAD5 downregulation exhibited opposite effects. Mechanistically, SPI1 could enhance SMAD5 transcription and expression. Downregulation of SMAD5 also reversed SPI1 overexpression-induced autophagy and osteogenic differentiation in MC3T3-E1 cells. In MC3T3-E1 cells under DEX stimulation, icariin increased SMAD5 expression by upregulating SPI1. Furthermore, icariin could attenuate SPI1 depletion-imposed inhibition of autophagy and osteogenic differentiation of MC3T3-E1 cells. Conclusion: Our findings demonstrate that the SPI1/SMAD5 cascade, with the ability to enhance osteogenic differentiation, underlies the promoting effect of icariin on osteogenic differentiation of MC3T3-E1 cells. In dexamethasone-stimulated murine pre-osteoblast MC3T3-E1 cells, icariin upregulates SPI1 and thus enhances SMAD5 transcription, leading to enhanced autophagy and osteogenic differentiation of MC3T3-E1 cells. [ABSTRACT FROM AUTHOR]

Cisplatin is a widely used chemotherapeutic drug that can induce ovarian damage. Icariin (ICA), a natural antioxidant derived from Epimedium brevicornum Maxim., has been found to protect against organ injury. The aim of the present study was to investigate whether ICA can exert an ovarian-protective effect on cisplatin induced premature ovarian failure (POF) and the underlying mechanism involved. The preventive effect of ICA was evaluated using body weight, the oestrous cycle, ovarian histological analysis, and follicle counting. ICA treatment increased body weight, ovarian weight, and the number of follicles and improved the oestrous cycle in POF mice. ICA reduced cisplatin-induced oxidative damage and upregulated the protein expression levels of Nrf2, GPX4 and HO-1. Moreover, ICA reduced the expression levels of Bax and γH2AX and inhibited ovarian apoptosis. In addition, ICA activated the Nrf2 pathway in vitro and reversed changes in the viability of cisplatin-induced KGN cells, reactive oxygen species (ROS) levels, lipid peroxidation, and apoptosis, and these effects were abrogated when Nrf2 was knocked down or inhibited. Molecular docking confirmed that ICA promotes the release of Nrf2 by competing with Nrf2 for binding to Keap1. The inhibitory effects of ICA on cisplatin-induced oxidative stress, ferroptosis, and apoptosis may be mediated by its modulatory effects on the Nrf2 pathway, providing a novel perspective on the potential mechanisms by which ICA prevents POF. [ABSTRACT FROM AUTHOR]

Introduction: Pancreatic cancer (PC) is a particularly aggressive malignancy with limited therapeutic options. The search for innovative treatments has focused on traditional Chinese medicine, specifically epimedium. This research investigates epimedium's active ingredients, potential targets, and underlying mechanisms in treating PC. Methods: High-performance liquid chromatography (HPLC) was used to quantify the active components of epimedium and HPLC-Q-TOF-MS was employed for qualitative identification. Potential targets of epimedium's active ingredients were identified using the TCMSP, ETCM, CTD, and Swiss Target Prediction databases. Potential PC-related targets were sourced from DisGeNET, GeneCards, and OMIM databases. A Venn diagram was utilized to identify overlapping PCrelated and epimedium targets. Core targets and pathways were elucidated through protein-protein interaction (PPI) network analysis, Gene Ontology (GO) assessments, and Reactome pathway enrichment analyses. Molecular docking techniques investigated interactions between active compounds and these targets. The expression and prognostic implications of target genes were evaluated using GEPIA2 and the Human Protein Atlas (HPA) databases. In vitro studies assessed the impact of epimedium extract (EPE) on Panc-1 cell viability, and Western blot analysis examined the expression levels of key targets. Results: Network pharmacological indicate that epimedium econtains active components such as baohuoside I, icariin, hyperoside, and epimedin B, which have potential therapeutic effects against PC. In vitro assays confirmed that EPE significantly reduced the viability of Panc-1 cells. Western blot analysis revealed a considerable decrease in the expression of key targets in EPE-treated cells, including AKT1, EGFR, p-EGFR, JUN, BCL2, IL6, and SRC. The R-HSA-1280215: Interleukin-4 and Interleukin-13 signaling pathways involving these genes were identified as potential therapeutic targets. Discussion: Epimedium holds promise as a candidate for treating PC. The modulation of interleukin-4 and interleukin-13 signaling pathways could be a pivotal mechanism by which epimedium impedes tumor development. Further research is warranted to validate these findings and explore the clinical applicability of epimedium in PC treatment. [ABSTRACT FROM AUTHOR]

Vascular dementia (VaD) has a serious impact on the patients' quality of life. Icariin (Ica) possesses neuroprotective potential for treating VaD, yet its oral bioavailability and blood–brain barrier (BBB) permeability remain challenges. This research introduced a PEG–PLGA-loaded chitosan hydrogel-based binary formulation tailored for intranasal delivery, enhancing the intracerebral delivery efficacy of neuroprotective agents. The formulation underwent optimization to facilitate BBB crossing, with examinations conducted on its particle size, morphology, drug-loading capacity, in vitro release, and biodistribution. Using the bilateral common carotid artery occlusion (BCCAO) rat model, the therapeutic efficacy of this binary formulation was assessed against chitosan hydrogel and PEG–PLGA nanoparticles loaded with Ica. Post-intranasal administration, enhanced cognitive function was evident in chronic cerebral hypoperfusion (CCH) rats. Further mechanistic evaluations, utilizing immunohistochemistry (IHC), RT-PCR, and ELISA, revealed augmented transcription of synaptic plasticity-associated proteins like SYP and PSD-95, and a marked reduction in hippocampal inflammatory markers such as IL-1β and TNF-α, highlighting the formulation's promise in alleviating cognitive impairment. The brain-derived neurotrophic factor (BDNF)/tropomyosin related kinase B (TrkB) pathway was activated significantly in the binary formulation compared with the other two. Our study demonstrates that the intranasal application of chitosan hydrogel loaded with Ica-encapsulated PEG–PLGA could effectively deliver Ica into the brain and enhance its neuroprotective effect. [ABSTRACT FROM AUTHOR]

Objective To investigate the effect of icariin on apoptosis of ovarian granulosa cells in rats with polycystic ovarian syndrome (PCOS) by regulating stromal cell-derived factor-1 (SDF-1)/CXC chemokine receptor-4 (CXCR4) signal axis. Methods Granular cells successfully isolated from ovarian tissue of rats in the PCOS model group were grouped into the PCOS group, the low-dose icariin group, the medium-dose icariin group, the high-dose icariin group, the Rr-SDF-1 group and the high-dose icariin+Rr-SDF-1 group. Granular cells successfully isolated from ovarian tissue of rats in the normal control group were taken as the control group. Enzyme-linked immunosorbent assay (ELISA) was applied to detect levels of follicle-stimulating hormone (FSH), testosterone (T) and luteinizing hormone (LH) in supernatant of ovarian granulosa cells. CCK-8 method and EdU staining were applied to detect granulosa cell proliferation. Flow cytometry was applied to detect granulosa cell apoptosis. Western blot assay was applied to detect the expression of Bcl-2 associated X (Bax), Cleaved aspartate-specific cysteine protease-3 (Cleaved Caspase-3), SDF-1, and CXCR4 proteins in granulosa cells. Results Compared with the control group, the FSH level, OD450 value and EdU positive cell rate were decreased in the PCOS group, and T and LH levels, apoptosis rate, Bax, Cleaved Caspase-3, SDF-1 and CXCR4 protein expression increased (P＜0.05). Compared with PCOS group, the FSH level, OD450 value and EdU positive cell rate were increased sequentially in the icariin low, medium and high dose groups, T and LH levels, apoptosis rate, Bax, Cleaved Caspase-3, SDF-1 and CXCR4 protein expression levels were decreased sequentially, and change trends of corresponding indicators were opposite to the above in the Rr-SDF-1 group (P＜0.05). Compared with the high-dose icariin group, FSH level, OD450 value and EdU positive cell rate were decreased in the high-dose icariin+Rr-SDF-1 group, and T and LH levels, apoptosis rate, Bax, Cleaved Caspase-3, SDF-1 and CXCR4 protein expression levels were increased (P＜0.05). Conclusion Icariin may inhibit the apoptosis of ovarian granulosa cells in PCOS rats by inhibiting the SDF-1/CXCR4 signaling pathway. [ABSTRACT FROM AUTHOR]

Background: Icariin (ICA) inhibits inflammatory response in various diseases, but the mechanism underlying ICA treating airway inflammation in asthma needs further understood. We aimed to predict and validate the potential targets of ICA against asthma‐associated airway inflammation using network pharmacology and experiments. Methods: The ovalbumin‐induced asthma‐associated airway inflammation mice model was established. The effects of ICA were evaluated by behavioral, airway hyperresponsiveness, lung pathological changes, inflammatory cell and cytokines counts. Next, the corresponding targets of ICA were mined via the SEA, CTD, HERB, PharmMapper, Symmap database and the literature. Pubmed‐Gene and GeneCards databases were used to screen asthma and airway inflammation‐related targets. The overlapping targets were used to build an interaction network, analyze gene ontology and enrich pathways. Subsequently, flow cytometry, quantitative real‐time PCR and western blotting were employed for validation. Results: ICA alleviated the airway inflammation of asthma; 402 targets of ICA, 5136 targets of asthma and 4531 targets of airway inflammation were screened; 216 overlapping targets were matched and predicted ICA possesses the potential to modulate asthmatic airway inflammation by macrophage activation/polarization. Additionally, ICA decreased M1 but elevated M2. Potential targets that were disrupted by asthma inflammation were restored by ICA treatment. Conclusions: ICA alleviates airway inflammation in asthma by inhibiting the M1 polarization of alveolar macrophages, which is related to metabolic reprogramming. Jun, Jak2, Syk, Tnf, Aldh2, Aldh9a1, Nos1, Nos2 and Nos3 represent potential targets of therapeutic intervention. The present study enhances understanding of the anti‐airway inflammation effects of ICA, especially in asthma. [ABSTRACT FROM AUTHOR]

Improving the reproductive ability and fertility of male ruminants is a central concern in animal husbandry. Phytogenic feed additives, known for their anti-inflammatory, antioxidant, and immunomodulatory properties, are commonly used in animal feed. Icariin (ICA), the primary flavonoid glucoside derived from Epimedium, is a traditional tonic in Chinese herbal medicine. However, its potential to enhance the reproductive performance of male ruminants remains unclear. In this study, twelve healthy adult male dairy goats were divided into two groups. The goats received oral administration of ICA at doses of 0 (control) and 50 mg/kg body weight daily for a consecutive period of 80 days during the breeding season. The effects of ICA on the reproductive performance was analyzed by histological examinations, semen quality analysis, and ELISA experiments. ELISA results showed a progressive increase in serum levels of GnIH, LH, and testosterone with the prolonged ICA treatment (p

Abstract Background Ischemic stroke presents a significant threat to human health due to its high disability rate and mortality. Currently, the clinical treatment drug, rt-PA, has a narrow therapeutic window and carries a high risk of bleeding. There is an urgent need to find new effective therapeutic drugs for ischemic stroke. Icariin (ICA), a key ingredient in the traditional Chinese medicine Epimedium, undergoes metabolism in vivo to produce Icaritin (ICT). While ICA has been reported to inhibit neuronal apoptosis after cerebral ischemia-reperfusion (I/R), yet its underlying mechanism remains unclear. Methods PC-12 cells were treated with 200 µM H2O2 for 8 h to establish a vitro model of oxidative damage. After administration of ICT, cell viability was detected by Thiazolyl blue tetrazolium Bromide (MTT) assay, reactive oxygen species (ROS) and apoptosis level, mPTP status and mitochondrial membrane potential (MMP) were detected by flow cytometry and immunofluorescence. Apoptosis and mitochondrial permeability transition pore (mPTP) related proteins were assessed by Western blotting. Middle cerebral artery occlusion (MCAO) model was used to establish I/R injury in vivo. After the treatment of ICA, the neurological function was scored by ZeaLonga socres; the infarct volume was observed by 2,3,5-Triphenyltetrazolium chloride (TTC) staining; HE and Nissl staining were used to detect the pathological state of the ischemic cortex; the expression changes of mPTP and apoptosis related proteins were detected by Western blotting. Results In vitro: ICT effectively improved H2O2-induced oxidative injury through decreasing the ROS level, inhibiting mPTP opening and apoptosis. In addition, the protective effects of ICT were not enhanced when it was co-treated with mPTP inhibitor Cyclosporin A (CsA), but reversed when combined with mPTP activator Lonidamine (LND). In vivo: Rats after MCAO shown cortical infarct volume of 32–40%, severe neurological impairment, while mPTP opening and apoptosis were obviously increased. Those damage caused was improved by the administration of ICA and CsA. Conclusions ICA improves cerebral ischemia-reperfusion injury by inhibiting mPTP opening, making it a potential candidate drug for the treatment of ischemic stroke. Graphical Abstract

Background: Ischaemic stroke can lead to many complications, but treatment options are limited. Icariin is a traditional Chinese medicine with reported neuroprotective effects against ischaemic cerebral injury; however, the underlying mechanisms by which icariin ameliorates cell apoptosis require further study. Purpose: This study aimed to investigate the therapeutic potential of icariin after ischaemic stroke and the underlying molecular mechanisms. Methods: N2a neuronal cells were used to create an in vitro oxygen-glucose deprivation (OGD) model. The effects of icariin on OGD cells were assessed using the CCK-8 kit to detect the survival of cells and based on the concentration, apoptosis markers, inflammation markers, and M2 pyruvate kinase isoenzyme (PKM2) expression were detected using western blotting, RT-qPCR, and flow cytometry. To investigate the underlying molecular mechanisms, we used the PKM2 agonist TEPP-46 and detected apoptosis-related proteins. Results: We demonstrated that icariin alleviated OGD-induced apoptosis in vitro. The expression levels of the apoptosis marker proteins caspase-3 and Bax were upregulated and Bcl-2 was downregulated. Furthermore, icariin reduced inflammation and downregulated the expression of PKM2. Moreover, activation of the PKM2 by pretreatment with the PKM2 agonist TEPP-46 enhanced the effects on OGD induced cell apoptosis in vitro. Conclusion: This study elucidated the underlying mechanism of PKM2 in OGD-induced cell apoptosis and highlighted the potential of icariin in the treatment of ischaemic stroke.

Abstract There is an increasing demand for innovative strategies that effectively promote osteogenesis and enhance bone regeneration. The critical process of bone regeneration involves the transformation of mesenchymal stromal cells into osteoblasts and the subsequent mineralization of the extracellular matrix, making up the complex mechanism of osteogenesis. Icariin’s diverse pharmacological properties, such as anti-inflammatory, anti-oxidant, and osteogenic effects, have attracted considerable attention in biomedical research. Icariin, known for its ability to stimulate bone formation, has been found to encourage the transformation of mesenchymal stromal cells into osteoblasts and improve the subsequent process of mineralization. Several studies have demonstrated the osteogenic effects of icariin, which can be attributed to its hormone-like function. It has been found to induce the expression of BMP-2 and BMP-4 mRNAs in osteoblasts and significantly upregulate Osx at low doses. Additionally, icariin promotes bone formation by stimulating the expression of pre-osteoblastic genes like Osx, RUNX2, and collagen type I. However, icariin needs to be effectively delivered to bone to perform such promising functions. Encapsulating icariin within nanoplatforms holds significant promise for promoting osteogenesis and bone regeneration through a range of intricate biological effects. When encapsulated in nanofibers or nanoparticles, icariin exerts its effects directly at the cellular level. Recalling that inflammation is a critical factor influencing bone regeneration, icariin's anti-inflammatory effects can be harnessed and amplified when encapsulated in nanoplatforms. Also, while cell adhesion and cell migration are pivotal stages of tissue regeneration, icariin-loaded nanoplatforms contribute to these processes by providing a supportive matrix for cellular attachment and movement. This review comprehensively discusses icariin-loaded nanoplatforms used for bone regeneration and osteogenesis, further presenting where the field needs to go before icariin can be used clinically.

Diabetes mellitus (DM) is a global health issue in the twenty-first century, and there are numerous challenges in preventing and alleviating its chronic complications. The herb Epimedium has beneficial therapeutic effects on various human diseases, including DM. Its major flavonoid component, icariin, has significant anti-DM activity and may help improve pancreatic β-cell dysfunction and insulin resistance. Furthermore, preclinical evidence has shown that icariin and its in vivo bioactive form, icariside II, have preventive and therapeutic effects on several diabetic complications, including diabetic cardiomyopathy, diabetic vascular endothelial disorder, diabetic nephropathy, and diabetic erectile dysfunction. In this review, we present the general and toxicological information concerning icariin and icariside II and review the anti-DM effects of icariin from a molecular perspective. Additionally, we discuss the potential benefits of icariin and icariside II on the important pathological mechanisms of various diabetic complications. Despite positive preclinical evidence, additional investigations are needed before relevant clinical studies can be conducted. Therefore, we conclude with suggestions for future research. Hopefully, this review will provide a comprehensive molecular perspective for future research and product development related to icariin and icariside II in treating DM and diabetic complications. [ABSTRACT FROM AUTHOR]

Parkinson’s disease (PD) is one of the most common neurodegenerative diseases characterized with a gradual loss of midbrain substantia nigra (SN) dopamine (DA) neurons. An excessive evidence demonstrated that microglia-mediated inflammation might be involved in the pathogenesis of PD. Thus, inhibition of neuroinflammation might possess a promising potential for PD treatment. Icariin (ICA), a single active component extracted from the Herba Epimedii, presents amounts of pharmacological properties, such as anti-inflammation, anti-oxidant, and anti-aging. Recent studies show ICA produced neuroprotection against brain dysfunction. However, the mechanisms underlying ICA-exerted neuroprotection are fully illuminated. In the present study, two different neurotoxins of 6-hydroxydopamine (6-OHDA) and lipopolysaccharide (LPS)-induced rat midbrain DA neuronal damage were applied to investigate the neuroprotective effects of ICA. In addition, primary rat midbrain neuron-glia co-cultures were performed to explore the mechanisms underlying ICA-mediated DA neuroprotection. In vitro data showed that ICA protected DA neurons from LPS/6-OHDA-induced DA neuronal damage and inhibited microglia activation and pro-inflammatory factors production via the suppression of nuclear factor-κB (NF-κB) pathway activation. In animal results, ICA significantly reduced microglia activation and significantly attenuated LPS/6-OHDA-induced DA neuronal loss and subsequent animal behavior changes. Together, ICA could protect DA neurons against LPS- and 6-OHDAinduced neurotoxicity both in vivo and in vitro. These actions might be closely associated with the inhibition of microglia-mediated neuroinflammation. [ABSTRACT FROM AUTHOR]

An ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was utilized to develop a technique for the simultaneous quantification of icariin and its primary metabolites in mouse urine. The levels of icariin, icariside Ⅰ, icariside Ⅱ, baohuoside Ⅱ, wushanicaritin, icaritin, and desmethylicaritin in mouse urine were analyzed subsequent to the oral administration of an icariin suspension. This study aimed to preliminarily investigate the excretion profile of icariin in mice. Using an aqueous solution containing 0.1% formic acid (A) and an acetonitrile solution containing 0.1% formic acid (B) as the mobile phases, icariin and its major metabolites demonstrated satisfactory linearity over the concentration range of 0.25–800 ng·mL−1 . The precision and accuracy of intra-day and inter-day measurements were all found to be within 15%. Seventy-two hours after the intragastric administration of icariin suspension to a mouse, the cumulative urinary excretion of icariin, icariside Ⅰ, icariside Ⅱ, baohuoside Ⅱ, wushanicaritin, icaritin, and desmethylicaritin was quantified as 13.48, 18.70, 2,627.51, 2.04, 10.04, 3,420.44, and 735.13 ng, respectively. The UPLC-MS/MS method developed in this research is characterized by its simplicity, sensitivity, and speed, making it well-suited for the concurrent quantification of icariin and its associated metabolites in urine. Additionally, it is appropriate for analyzing urine samples that may contain multiple drugs in future investigations. [ABSTRACT FROM AUTHOR]

Background: Ischemic stroke presents a significant threat to human health due to its high disability rate and mortality. Currently, the clinical treatment drug, rt-PA, has a narrow therapeutic window and carries a high risk of bleeding. There is an urgent need to find new effective therapeutic drugs for ischemic stroke. Icariin (ICA), a key ingredient in the traditional Chinese medicine Epimedium, undergoes metabolism in vivo to produce Icaritin (ICT). While ICA has been reported to inhibit neuronal apoptosis after cerebral ischemia-reperfusion (I/R), yet its underlying mechanism remains unclear. Methods: PC-12 cells were treated with 200 µM H2O2 for 8 h to establish a vitro model of oxidative damage. After administration of ICT, cell viability was detected by Thiazolyl blue tetrazolium Bromide (MTT) assay, reactive oxygen species (ROS) and apoptosis level, mPTP status and mitochondrial membrane potential (MMP) were detected by flow cytometry and immunofluorescence. Apoptosis and mitochondrial permeability transition pore (mPTP) related proteins were assessed by Western blotting. Middle cerebral artery occlusion (MCAO) model was used to establish I/R injury in vivo. After the treatment of ICA, the neurological function was scored by ZeaLonga socres; the infarct volume was observed by 2,3,5-Triphenyltetrazolium chloride (TTC) staining; HE and Nissl staining were used to detect the pathological state of the ischemic cortex; the expression changes of mPTP and apoptosis related proteins were detected by Western blotting. Results: In vitro: ICT effectively improved H2O2-induced oxidative injury through decreasing the ROS level, inhibiting mPTP opening and apoptosis. In addition, the protective effects of ICT were not enhanced when it was co-treated with mPTP inhibitor Cyclosporin A (CsA), but reversed when combined with mPTP activator Lonidamine (LND). In vivo: Rats after MCAO shown cortical infarct volume of 32–40%, severe neurological impairment, while mPTP opening and apoptosis were obviously increased. Those damage caused was improved by the administration of ICA and CsA. Conclusions: ICA improves cerebral ischemia-reperfusion injury by inhibiting mPTP opening, making it a potential candidate drug for the treatment of ischemic stroke. [ABSTRACT FROM AUTHOR]

It was reported that the Wnt/β‐catenin pathway is involved in the regulation of aerobic glycolysis and that brain glycolytic dysfunction results in the development of Alzheimer's disease (AD). Icariin (ICA), an active component extracted from Epimedii Folium, has been reported to produce neuroprotective effects in multiple models of AD, but its underlying mechanism remains to be fully described. We aimed to investigate the protective effects of ICA on animal and cell models of AD and confirm whether the Wnt/β‐catenin pathway has functions in the neuroprotective function of ICA. The 3 × Tg‐AD mice were treated with ICA. HT22 cells, the Aβ25‐35 peptide and Dickkopf‐1 (DKK1) agent (a specific inhibitor of the Wnt/β‐catenin pathway) were used to further explore the underlying mechanism of ICA that produces anti‐AD effects. Behavioral examination, western blotting assay, staining analysis, biochemical test, and lactate dehydrogenase (LDH) assays were applied. We first demonstrated that ICA significantly improved cognitive function and autonomous behavior, reduced neuronal damage, and reversed the protein levels and activities of glycolytic key enzymes, and expression of protein molecules of the canonical Wnt signaling pathway, in 3 × Tg‐AD mice back to wild‐type levels. Next, we further found that ICA increased cell viability and effectively improved the dysfunctional glycolysis in HT22 cells injured by Aβ25‐35. However, when canonical Wnt signaling was inhibited by DKK1, the above effects of ICA on glycolysis were abolished. In summary, ICA exerts neuroprotective effects in 3 × Tg‐AD animals and AD cellular models by enhancing the function of glycolysis through activation of the Wnt/β‐catenin pathway. [ABSTRACT FROM AUTHOR]

The experiment was designed to explore the internal mechanisms of produced Epimedium by mutton tallow. Taking Epimedium from Gansu as the research subject, it was processed with mutton tallow, beef tallow and lard fat, respectively. The content of the effective components were extracted and determined by UV and HPLC. The contents of flavonoids, icariin, baohuoside Ⅰ, and epimedin C in different processed products and unprocessed products were compared. The results were as follows: the flavonoids content of processed products was significantly lower than that of unprocessed products (6.84%). The flavonoids content of processed products by mutton tallow (5.69%) was significantly higher than that of other processed products. There was no significant difference in the content of icariin among unprocessed products and processed products by mutton tallow and beef tallow. The content of icariin in the mutton tallow processed products and beef tallow processed products was 0.77%, 0.70%. The content of icariin in the unprocessed products was 0.70%. The content of icariin in the pork fat processed products was 0.50%, which was significantly lower than that in the unprocessed products and other processed products. There was no significant difference in the content of baohuoside Ⅰ between the mutton tallow processed products and beef tallow processed products (0.08% and 0.07%), but the content of baohuoside Ⅰ was significantly higher than that of unprocessed products (0.05%) and processed products by pork fat (0.04%). The con‐tent of epimedin C in processed products by mutton tallow, beef tallow and pork fat was 0.29%, 0.27% and 0.24%, which was significantly lower than that in unprocessed products (0.57%). It was concluded that the best excipients of epimedium were muttontallow, followed by beef tallow. [ABSTRACT FROM AUTHOR]

The treatment of bone defects has always been a major problem in orthopedics. Due to the limited regenerative capacity of bone, large-scale bone defects often cannot be healed by themselves. Icariin (ICA) has an obvious effect on bone formation. ICA contained in Icariin is a flavonoid compound and an active compound, which has dual activities of promoting bone formation and inhibiting bone absorption. Icariin can also be delivered locally through a variety of biological materials to promote bone defect repair, showing a good bone induction potential. It has been found that ICA mainly repairs bone defects by promoting bone formation, cartilage formation, angiogenesis, and by inhibiting bone resorption and anti-inflammatory effects. In this paper, ICA repair of bone defect and its mechanism of action are summarized in order to provide some reference for the follow-up research. [ABSTRACT FROM AUTHOR]

This article discusses the effects of icariin, a compound found in certain plants, on insulin resistance in muscle cells. The study found that icariin treatment protected muscle cells against insulin resistance induced by a fatty acid called PA. This protection was mediated by the protein TXNIP, which was reduced by icariin through a mechanism involving the proteasome system. Icariin also reduced ER stress, inflammation, and phosphorylation of STAT3, which are all associated with insulin resistance. However, when the proteasome system was inhibited, the beneficial effects of icariin were abolished. The study suggests that icariin may be a potential therapeutic option for preventing and treating insulin resistance and diabetes. However, the study has some limitations and further research is needed to confirm these findings. [Extracted from the article]

Deer bone in traditional Chinese medicine is valued for its unique biomimetic bone matrix components. Icariin (Ica), which is claimed by Chinese medicine as a bone-repairing agent, is also widely accepted. In this report, a novel Icaencapsulated PLGA/nano deer bone meal (Ica@PLGA/nBM) biomimetic microsphere was fabricated. The Ica could be released from microspheres in a controlled manner. In vitro results showed that the microsphere had good biocompatibility due to the addition of nBM and the MC3T3-E1 cells could adhere to and grow well on the microspheres. Moreover, Ica and nBM in microspheres could synergically promote the ALP activity of MC3T3-E1 cells significantly (p < 0.05), which indicated the acceleration of osteogenic differentiation. The RT-PCR results showed that the expression levels of bone matrix proteins (Col1 & OPN) were enhanced by the composite (p < 0.05). In addition, Ica and nBM in microspheres also obviously promoted the expression of RUNX2 and BMP-2, respectively (p < 0.05). The in vivo results demonstrated the effective promotion of Ica@PLGA/nBM on complete bony connection to cover the defect site and the bone defect repair. And the Ica(M)@PLGA/nBM microspheres provided the best repair results. All the results confirmed that the Ica@PLGA/nBM composite biomimetic microsphere has the potential for clinical application. [ABSTRACT FROM AUTHOR]

Tubulointerstitial fibrosis is an inevitable consequence of all progressive chronic kidney disease (CKD) and contributes to a substantial health burden worldwide. Icariin, an active flavonoid glycoside obtained from Epimedium species, exerts potential antifibrotic effect. The study aimed to explore the protective effects of icariin against tubulointerstitial fibrosis in unilateral ureteral obstruction (UUO)‐induced CKD mice and TGF‐β1‐treated HK‐2 cells, and furthermore, to elucidate the underlying mechanisms. The results demonstrated that icariin significantly improved renal function, alleviated tubular injuries, and reduced fibrotic lesions in UUO mice. Furthermore, icariin suppressed renal inflammation, reduced oxidative stress as evidenced by elevated superoxide dismutase activity and decreased malondialdehyde level. Additionally, TOMM20 immunofluorescence staining and transmission electron microscope revealed that mitochondrial mass and morphology of tubular epithelial cells in UUO mice was restored by icariin. In HK‐2 cells treated with TGF‐β1, icariin markedly decreased profibrotic proteins expression, inhibited inflammatory factors, and protected mitochondria along with preserving mitochondrial morphology, reducing reactive oxygen species (ROS) and mitochondrial ROS (mtROS) overproduction, and preserving membrane potential. Further investigations demonstrated that icariin could activate nuclear factor erythroid 2‐related factor 2 (Nrf2)/heme oxygenase‐1 (HO‐1) pathway both in vivo and in vitro, whereas inhibition of Nrf2 by ML385 counteracted the protective effects of icariin on TGF‐β1‐induced HK‐2 cells. In conclusion, icariin protects against renal inflammation and tubulointerstitial fibrosis at least partly through Nrf2‐mediated attenuation of mitochondrial dysfunction, which suggests that icariin could be developed as a promising therapeutic candidate for the treatment of CKD. Significance statement: Tubulointerstitial fibrosis is an unavoidable consequence of advancing chronic kidney disease (CKD) and imposes a substantial global health burden. However, few effective therapeutic agents are available at present. The present study investigated the ameliorative effects of icariin on tubulointerstitial fibrosis (TIF) both in vivo and in vitro, and demonstrated that icariin inhibited inflammation and oxidative stress, at least partially, through improving Nrf2 activity and subsequent mitochondrial function, and eventually mitigated TIF and preserved renal function, suggesting that icariin could be developed as a promising therapeutic candidate for the treatment of CKD. [ABSTRACT FROM AUTHOR]

Icaritin is a natural prenylated flavonoid derived from the Chinese herb Epimedium. The compound has shown antitumor effects in various cancers, especially hepatocellular carcinoma (HCC). Icaritin exerts its anticancer activity by modulating multiple signaling pathways, such as IL‐6/JAK/STAT3, ER‐α36, and NF‐κB, affecting the tumor microenvironment and immune system. Several clinical trials have evaluated the safety and efficacy of icaritin in advanced HCC patients with poor prognoses, who are unsuitable for conventional therapies. The results have demonstrated that icaritin can improve survival, delay progression, and produce clinical benefits in these patients, with a favorable safety profile and minimal adverse events. Moreover, icaritin can enhance the antitumor immune response by regulating the function and phenotype of various immune cells, such as CD8+ T cells, MDSCs, neutrophils, and macrophages. These findings suggest that icaritin is a promising candidate for immunotherapy in HCC and other cancers. However, further studies are needed to elucidate the molecular mechanisms and optimal dosing regimens of icaritin and its potential synergistic effects with other agents. Therefore, this comprehensive review of the scientific literature aims to summarize advances in the knowledge of icaritin in preclinical and clinical studies as well as the pharmacokinetic, metabolism, toxicity, and mechanisms action to recognize the main challenge, gaps, and opportunities to develop a medication that cancer patients can use. Thus, our main objective was to clarify the current state of icaritin for use as an anticancer drug. [ABSTRACT FROM AUTHOR]

Icariin has shown the potential to treat osteoarthritis (OA), but the specific mechanism still needs further exploration. Therefore, this study attempted to reveal the effect and mechanism of icariin on OA based on in vitro and in vivo experiments. In vivo, a mouse model of OA was established by cutting the anterior cruciate ligament, and 10 mg/kg icariin was given to mice orally. Then, the OA injury and pathological changes of cartilage tissue in mice were identified by OA index and hematoxylin and eosin staining. In vitro, the viability of C28/I2 cells incubated with different concentrations of icariin was detected by 3‐(4,5)‐dimethylthiahiazo (‐z‐y1)‐3,5‐di‐ phenytetrazoliumromide assay. Subsequently, C28/I2 cells induced by IL‐1β were used as the cell model of OA, the expression of Sirtuin (SIRT)‐1 in cells was knocked down, and icariin was added for intervention. Next, western blot was used to observe the expression level of sirtuin 1 (SIRT‐1)‐Nrf2‐heme oxygenase 1 (HO‐1) signaling pathway‐related proteins in cells of each group. Besides, cell viability and apoptosis were detected by MTT and apoptosis assay, and DNA damage was observed by comet assay. In vivo experiments, intragastric administration of icariin could effectively reduce the OA index of mice, improve the pathological changes of cartilage tissue, and obviously activated the SIRT‐1‐Nrf2‐HO‐1 signaling pathway. In vitro experiments, icariin did not exhibit toxic effect on C28/I2 cells, but could activate the SIRT‐1‐Nrf2‐HO‐1 signaling pathway, improve the viability, reduce the level of apoptosis and relieve the DNA damage in OA cells; however, these effects were inhibited by si‐ SIRT‐1. Icariin can improve the symptoms of OA by activating the SIRT‐1‐Nrf2‐HO‐1 signaling pathway. [ABSTRACT FROM AUTHOR]

Icariin, a flavonoid glycoside, is extracted from Epimedium. This study aimed to investigate the vascular protective effects of icariin in type 1 diabetic rats by inhibiting high-mobility group box 1 (HMGB1)-related inflammation and exploring its potential mechanisms. The impact of icariin on vascular dysfunction was assessed in streptozotocin (STZ)-induced diabetic rats through vascular reactivity studies. Western blotting and immunofluorescence assays were performed to measure the expressions of target proteins. The release of HMGB1 and pro-inflammation cytokines were measured by enzyme-linked immunosorbent assay (ELISA). The results revealed that icariin administration enhanced acetylcholine-induced vasodilation in the aortas of diabetic rats. It also notably reduced the release of pro-inflammatory cytokines, including interleukin-8 (IL-8), IL-6, IL-1β, and tumor necrosis factor-alpha (TNF-α) in diabetic rats and high glucose (HG)-induced human umbilical vein endothelial cells (HUVECs). The results also unveiled that the pro-inflammatory cytokines in the culture medium of HUVECs could be increased by rHMGB1. The increased release of HMGB1 and upregulated expressions of HMGB1-related inflammatory factors, including advanced glycation end products (RAGE), Toll-like receptor 4 (TLR4), and phosphorylated p65 (p-p65) in diabetic rats and HG-induced HUVECs, were remarkably suppressed by icariin. Notably, HMGB1 translocation from the nucleus to the cytoplasm in HUVECs under HG was inhibited by icariin. Meanwhile, icariin could activate G protein-coupled estrogen receptor (GPER) and sirt1. To explore the role of GPER and Sirt1 in the inhibitory effect of icariin on HMGB1 release and HMGB-induced inflammation, GPER inhibitor and Sirt1 inhibitor were used in this study. These inhibitors diminished the effects of icariin on HMGB1 release and HMGB1-induced inflammation. Specifically, the GPER inhibitor also negated the activation of Sirt1 by icariin. These findings suggest that icariin activates GPER and increases the expression of Sirt1, which in turn reduces HMGB1 translocation and release, thereby improving vascular endothelial function in type 1 diabetic rats by inhibiting inflammation. [ABSTRACT FROM AUTHOR]

A three‐dimensional alginate‐coated scaffold (GAIS) was constructed in the present study to showcase the multidifferentiation potential of peripheral blood mesenchymal stem cells (PBMSCs) and to investigate the role and mechanism by which Icariin (ICA)/stromal cell‐derived factor (SDF‐1α)/PBMSCs promote damaged articular repair. In addition, the ability of ICA, in combination with SDF‐1α, to promote the migration and proliferation of stem cells was validated through the utilization of CCK‐8 and migration experiments. The combination of ICA and SDF‐1α inhibited the differentiation of PBMSCs into cartilage, as demonstrated by in vivo experiments and histological staining. Both PCR and western blot experiments showed that GAIS could upregulate the expression of particular genes in chondrocytes. In comparison to scaffolds devoid of alginate (G0), PBMSCs seeded into GAIS scaffolds exhibited a greater rate of proliferation, and the conditioned medium derived from scaffolds containing SDF‐1α enhanced the capacity for cell migration. Moreover, after a 12‐week treatment period, GAIS, when successfully transplanted into osteochondral defects of mice, was found to promote cartilage regeneration and repair. The findings, therefore, demonstrate that GAIS enhanced the in vitro capabilities of PBMSCs, including proliferation, migration, homing and chondrogenic differentiation. In addition, ICA and SDF‐1α effectively collaborated to support cartilage formation in vivo. Thus, the ICA/SDF‐1α/PBMSC‐loaded biodegradable alginate‐gelatin scaffolds showcase considerable potential for use in cartilage repair. [ABSTRACT FROM AUTHOR]

Objective: The objectives of this study were to define the superiority of icariin and its derivatives' anti-inflammatory activities and to create a reference framework for evaluating preclinical evidence. This method combines machine learning and meta-analysis to identify underlying biological pathways. Methods: Data came from PubMed, Embase, Web of Science, and the Cochrane Library. SYRCLE was used to evaluate the risk of bias in a subset of research. Meta-analysis and detailed subgroup analyses, categorized by species, genders, disease type, dosage, and treatment duration, were performed using R and STATA 15.0 software to derive nuanced insights. Employing R software (version 4.2.3) and the tidymodels package, the analysis focused on constructing a model and selecting features, with TNF-α as the dependent variable. This approach aims to identify significant predictors of drug efficacy. An in-depth literature facilitated the synthesis of anti-inflammatory mechanisms attributed to icariin and its constituent compounds. Results: Following a meticulous search and selection process, 19 studies, involving 370 and 260 animals were included in the meta-analysis and machine-learning assessment, respectively. The findings revealed that icariin and its derivatives markedly reduced inflammation markers, including TNF-α and IL-1β. Additionally, machine-learning outcomes, with TNF-α as the target variable, indicated enhanced anti-inflammatory effects of icariin across respiratory, urological, neurological, and digestive disease types. These effects were more pronounced at doses exceeding 27.52 mg/kg/day and treatment durations beyond 31.22 days. Conclusion: Strong anti-inflammatory effects are exhibited by icariiin and its derivatives, which are especially beneficial in the management of digestive, neurological, pulmonary, and urinary conditions. Effective for periods longer than 31.22 days and at dosages more than 27.52 mg/kg/day. Subsequent research will involve more targeted animal experiments and safety assessments to obtain more comprehensive preclinical evidence.

Icariin (ICA), a natural flavonoid glucoside from traditional Chinese medicine, possesses various pharmacological properties such as anti-inflammatory, anti-aging, and neuroprotective effects. Recent studies suggest its potential in treating Alzheimer’s disease (AD). However, the exact mechanisms of how ICA modulates neuroinflammation in AD remain unclear. In this study, oral ICA administration improved cognitive function in mice, decreasing escape latency in behavioral tests and altering protein levels related to AD pathology, including boosting acetylcholine and reducing p-tau/tau and acetylcholinesterase. Additionally, in 3 × Tg-AD mice, ICA therapy inhibited microglia and astrocyte activation and reduced inflammatory cytokines (IL-1β, TNF-α, IL-6) at the protein level. RNA-seq analysis revealed decreased expression of Nrxn3, Meg3, and Malat1 genes in 3 × Tg-AD animals treated with ICA. Furthermore, ICA activated the Akt/GSK-3β signaling pathway, known for its role in neuroinflammation, suggesting a potential mechanism by which ICA suppresses inflammation. This study proposes Meg3 and Malat1 lncRNA as therapeutic targets against AD, offering a novel approach for combating neuroinflammation in AD through the inhibition of the Akt/GSK-3β pathway.

Bovine viral diarrhoea virus (BVDV) can cause diarrhoea (BVD) in an animal herd, leading to heavy economic losses. There are limited drugs available for treating and controlling BVD. This research aims to investigate the antiviral and immunoregulatory effects of two traditional Chinese herb extracts against BVDV infection. The extracts are matrine and icariin, which have been proved to have immunostimulant and antiviral effects.