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Objective To investigate the effects of pinocembrin（PIN）on myocardial inflammation and stimulator of interferon gene / TANK-binding kinase 1 / interferon regulatory factor 3（STING / TBK1 / IRF3）signaling pathway in diabetic cardiomyopathy（DCM）rats. Methods A DCM rat model was constructed，and the modeled rats were separated into a model group，PIN low- and high- dose groups，and a PIN high-dose + STING pathway activator DMXAA group （high-dose of PIN + DMXAA group），with 12 rats in each group. Twelve normal rats were taken as a control group. Cardiac function and glucose metabolism levels were detected. ELISA was used to detect the level of inflammationrelated factors and the level of myocardial injury-related indexes. HE staining was used to observe the pathological changes of myocardial tissue. Masson staining was used to observe the degree of myocardial tissue fibrosis. The expression of fibrosis-related protein lysyl oxidase（LOX）in myocardial tissue was detected by immunohistochemistry. Western Blot was used to detect the expression of proteins related to STING / TBK1 / IRF3 signaling pathway. Results Myocardial tissue structure and cellular arrangement were disordered in the DCM group compared with the control group， myocardial fibers were broken and the gap was widened， cardiomyocytes were hypertrophied with obvious inflammatory cell infiltration， blue collagen fibers were increased， and the percentage of myocardial collagenous fibrosis was elevated. The LVEF，LVFS，Em/Am values decreased. The levels of FBG，FINS，CK-Mb， cTnI， TNF - α， IL-6， IL-1β and the expression of LOX， p-STING / STING， p-TBK1 / TBK1 and p-IRF3 / IRF3 elevated（P < 0.05）. Myocardial tissue structure and cell arrangement were relatively neat in PIN low-and high-dose groups compared with the DCM group，the breakage of myocardial fibers and the amplification of interstitial space were reduced， the morphology of myocardial cells tended to be normal， the infiltration of inflammatory cells was insignificant，and the blue collagen fibers were reduced，the percentage of myocardial collagen fibrosis decreased. The LVEF，LVFS，Em/Am values increased，the levels of FBG，FINS，CK-Mb，cTnI，TNF-α，IL-6，IL-1β and the expression of LOX， p-STING / STING， p-TBK1 / TBK1， p-IRF3 / IRF3 decreased（P < 0.05）. Myocardial tissue damage was more severe in the high-dose of PIN+DMXAA group than in PIN high-dose group，with increased fibrosis， inflammatory cell infiltration and blue collagen fibers. The percentage of myocardial collagen fibrosis elevated. The LVEF，LVFS，Em/Am values decreased. The levels of FBG，FINS，CK-Mb，cTnI，TNF-α，IL-6，IL-1β and the expression of LOX，p-STING/STING，p-TBK1/TBK1，p-IRF3/IRF3 elevated（P < 0.05）. Conclusion Pinocembrin ameliorates myocardial inflammation in diabetic cardiomyopathy rats. Its mechanism of action is related to the inhibition of STING/TBK1/IRF3 signaling pathway. [ABSTRACT FROM AUTHOR]

After stroke, there is a high incidence of acute lung injury and impairment of intestinal barrier function. In this research, the effects of pinocembrin on organ injuries induced by cerebral ischemia–reperfusion were investigated in mice with middle cerebral artery occlusion/reperfusion (MCAO/R) and further explored the possible mechanism. The potential targets of pinocembrin against MCAO/R were obtained by online tools. An MCAO/R model was developed in C57BL/6 J mice, in combination with pinocembrin administration and lentivirus-mediated gene intervention. Pinocembrin alleviated neurological impairment, reduced the volume of cerebral infarction, attenuated pathological injury of brain tissues in MCAO/R-induced mice by promoting the expression of dipeptidyl peptidase 9 (DPP9), and blocked the nucleotide‐binding domain leucine‐rich repeat pyrin domain containing 1 (NLRP1) inflammasome activation. Moreover, pinocembrin attenuated the infiltration of inflammatory cells in the lungs and intestinal histopathological injury induced by MCAO/R. The above effects of pinocembrin were reversed by knocking down DPP9. These findings indicated that pinocembrin inhibits NLRP1 inflammasome activation by inducing DPP9, thus mitigating brain, lung, and intestinal injuries induced by MCAO/R. [ABSTRACT FROM AUTHOR]

Abstract Background Flavonoids are a structurally diverse group of secondary metabolites, predominantly produced by plants, which include a range of compounds with pharmacological importance. Pinocembrin is a key branch point intermediate in the biosynthesis of a wide range of flavonoid subclasses. However, replicating the biosynthesis of these structurally diverse molecules in heterologous microbial cell factories has encountered challenges, in particular the modest pinocembrin titres achieved to date. In this study, we combined genome engineering and enzyme candidate screening to significantly enhance the production of pinocembrin and its derivatives, including chrysin, pinostrobin, pinobanksin, and galangin, in Escherichia coli. Results By implementing a combination of established strain engineering strategies aimed at enhancing the supply of the building blocks phenylalanine and malonyl-CoA, we constructed an E. coli chassis capable of accumulating 353 ± 19 mg/L pinocembrin from glycerol, without the need for precursor supplementation or the fatty acid biosynthesis inhibitor cerulenin. This chassis was subsequently employed for the production of chrysin, pinostrobin, pinobanksin, and galangin. Through an enzyme candidate screening process involving eight type-1 and five type-2 flavone synthases (FNS), we identified Petroselinum crispum FNSI as the top candidate, producing 82 ± 5 mg/L chrysin. Similarly, from a panel of five flavonoid 7-O-methyltransferases (7-OMT), we found pinocembrin 7-OMT from Eucalyptus nitida to yield 153 ± 10 mg/L pinostrobin. To produce pinobanksin, we screened seven enzyme candidates exhibiting flavanone 3-hydroxylase (F3H) or F3H/flavonol synthase (FLS) activity, with the bifunctional F3H/FLS enzyme from Glycine max being the top performer, achieving a pinobanksin titre of 12.6 ± 1.8 mg/L. Lastly, by utilising a combinatorial library of plasmids encoding G. max F3H and Citrus unshiu FLS, we obtained a maximum galangin titre of 18.2 ± 5.3 mg/L. Conclusion Through the integration of microbial chassis engineering and screening of enzyme candidates, we considerably increased the production levels of microbially synthesised pinocembrin, chrysin, pinostrobin, pinobanksin, and galangin. With the introduction of additional chassis modifications geared towards improving cofactor supply and regeneration, as well as alleviating potential toxic effects of intermediates and end products, we anticipate further enhancements in the yields of these pinocembrin derivatives, potentially enabling greater diversification in microbial hosts.

Objective: To investigate the influences of Pinocembrin on inflammatory injury in rats with acute myocardial infarction (AMI) by regulating TLR4/NF-κB/NLRP3 signaling pathway. Methods: The AMI model was established by coronary ligation, and the rats were grouped into Sham group, AMI group, Pinocembrin group (5 mg/kg tail vein injection), TLR4 inhibitor group (TAK-242 group, 2.0 mg/kg tail vein injection), the levels of cardiac function indexes (LVEF, LVEDD, LVESD, FS) and serum LDH, cTnⅠ, IL-6, IL-β and TNF-α were detected in rats, TTC staining, HE staining and Masson staining were applied to observe myocardial infarction and myocardial histopathological changes in rats, cardiomyocyte apoptosis was detected by TUNEL method, immunohistochemistry and Western blot were applied to detect TLR4/NF-κB/NLRP3 pathway-related proteins in rat myocardial tissue. Results: Compared with Sham group, the myocardial infarction area increased, the number of myocardial cells decreased, some myocardial fibers were broken, inflammatory cells infiltrated, collagen fibers increased, and the apoptosis rate was obviously increased in AMI group (P<0.05), LVEDD, LVESD, serum LDH, cTnⅠ, IL-6, IL-β, TNF-α levels, myocardial tissue TLR4, MyD88, p-NF-κB p65, NLRP3, Caspase-1 expression levels were obviously increased (P<0.05), while LVEF and FS were obviously decreased (P<0.05); compared with AMI group, the myocardial infarction area of the Pinocembrin group and the TAK-242 group were reduced, the cell damage and inflammatory infiltration were reduced, the necrotic cells were obviously reduced, and the apoptosis rate was obviously reduced (P<0.05), LVEDD, LVESD, serum LDH, cTnⅠ, IL-6, IL-β, TNF-α levels, myocardial tissue TLR4, MyD88, p-NF-κB p65, NLRP3, Caspase-1 expression levels were decreased (P<0.05), LVEF and FS were obviously increased (P<0.05); there was no obvious difference in each index between Pinocembrin group and TAK-242 group(P>0.05). Conclusion: Pinocembrin may attenuate myocardial inflammatory injury caused by AMI by inhibiting TLR4/NF-κB/NLRP3 signaling pathway. [ABSTRACT FROM AUTHOR]

Background: Flavonoids are a structurally diverse group of secondary metabolites, predominantly produced by plants, which include a range of compounds with pharmacological importance. Pinocembrin is a key branch point intermediate in the biosynthesis of a wide range of flavonoid subclasses. However, replicating the biosynthesis of these structurally diverse molecules in heterologous microbial cell factories has encountered challenges, in particular the modest pinocembrin titres achieved to date. In this study, we combined genome engineering and enzyme candidate screening to significantly enhance the production of pinocembrin and its derivatives, including chrysin, pinostrobin, pinobanksin, and galangin, in Escherichia coli. Results: By implementing a combination of established strain engineering strategies aimed at enhancing the supply of the building blocks phenylalanine and malonyl-CoA, we constructed an E. coli chassis capable of accumulating 353 ± 19 mg/L pinocembrin from glycerol, without the need for precursor supplementation or the fatty acid biosynthesis inhibitor cerulenin. This chassis was subsequently employed for the production of chrysin, pinostrobin, pinobanksin, and galangin. Through an enzyme candidate screening process involving eight type-1 and five type-2 flavone synthases (FNS), we identified Petroselinum crispum FNSI as the top candidate, producing 82 ± 5 mg/L chrysin. Similarly, from a panel of five flavonoid 7-O-methyltransferases (7-OMT), we found pinocembrin 7-OMT from Eucalyptus nitida to yield 153 ± 10 mg/L pinostrobin. To produce pinobanksin, we screened seven enzyme candidates exhibiting flavanone 3-hydroxylase (F3H) or F3H/flavonol synthase (FLS) activity, with the bifunctional F3H/FLS enzyme from Glycine max being the top performer, achieving a pinobanksin titre of 12.6 ± 1.8 mg/L. Lastly, by utilising a combinatorial library of plasmids encoding G. max F3H and Citrus unshiu FLS, we obtained a maximum galangin titre of 18.2 ± 5.3 mg/L. Conclusion: Through the integration of microbial chassis engineering and screening of enzyme candidates, we considerably increased the production levels of microbially synthesised pinocembrin, chrysin, pinostrobin, pinobanksin, and galangin. With the introduction of additional chassis modifications geared towards improving cofactor supply and regeneration, as well as alleviating potential toxic effects of intermediates and end products, we anticipate further enhancements in the yields of these pinocembrin derivatives, potentially enabling greater diversification in microbial hosts. [ABSTRACT FROM AUTHOR]

Propolis, such as is used as bio-cosmetics and in functional materials, is increasing because of its antioxidant medicinal benefits. However, its pharmacological and chemical composition is highly variable, relative to its geography and botanical origins. Comparative studies on three propolis samples collected from different regions in Korea have been essential for linking its provenance, chemical composition, and biological activity, thereby ensuring the efficient utilization of its beneficial properties. Here, we report the chemical composition and biological activities such as the antioxidant, wound healing, and anti-inflammatory effects of the ethanolic extract of Korean propolis collected from two regions. We compared the chemical constituents of three 70% ethanol-extracted (EE) samples, including the Andong, Gongju field (GF), and Gongju mountain (GM)-sourced propolis using Gas chromatography–mass spectrometry (GC–MS). The major and common components of these EE Korean propolis were flavonoids such as pinocembrin (12.0–17.7%), chrysin (5.2–6.8%), and apigenin (5.30–5.84%). The antioxidant property using a 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity assay of EEP showed substantial differences among samples with the highest from Andong. The sample 10% GM levigated in simple ointment was found to be the most active in wound healing activity based on the excision, incision, and dead space wound models. The potential of propolis for wound healing is mainly due to its evidenced properties, such as its antimicrobial, anti-inflammatory, analgesic, and angiogenesis promoter effects, which need further study. The anti-inflammatory activity and NO production inhibitory effect were highest in GM samples. However, GM and GF samples demonstrated similar inhibitory effects on the expression of inflammatory mediators such as iNOS, IL-1β, and IL-6. The presence of a higher concentration of flavonoids in Korean EE propolis might be responsible for their promising wound healing, anti-inflammatory, and antioxidant properties. [ABSTRACT FROM AUTHOR]

Non-small cell lung cancer (NSCLC), representing 85% of lung cancer cases, is characterized by its heterogeneity and progression through distinct stages. This study applied Weighted Gene Coexpression Network Analysis (WGCNA) to explore the molecular mechanisms of NSCLC and identify potential therapeutic targets. Gene expression data from the GEO database were analyzed across four NSCLC stages (NSCLC1, NSCLC2, NSCLC3, and NSCLC4), with the NSCLC2 dataset selected as the reference for module preservation analysis. WGCNA identified eight highly preserved modules— Cyan, Yellow, Red, Dark Turquoise, Turquoise, White, Purple, and Royal Blue—across datasets, which were enriched in key pathways such as “Cell cycle” and “Pathways in cancer”, involving processes like cell division and inflammatory responses. Hub genes, including PLK1, CDK1, and EGFR, emerged as critical regulators of tumor proliferation and immune responses. Estrogen receptor ESR1 was also highlighted, correlating with improved survival outcomes, suggesting its potential as a prognostic marker. Signature-based drug repurposing analysis identified promising therapeutic candidates, including GW-5074, which inhibits RAF and disrupts the EGFR–RAS–RAF–MEK–ERK signaling cascade, and olomoucine, a CDK1 inhibitor. Additional candidates like pinocembrin, which reduces NSCLC cell invasion by modulating epithelial-mesenchymal transition, and citalopram, an SSRI with anti-carcinogenic properties, were also identified. These findings provide valuable insights into the molecular underpinnings of NSCLC and suggest new directions for therapeutic strategies through drug repurposing. [ABSTRACT FROM AUTHOR]

Knowledge of the chemical composition of propolis is crucial for understanding the characteristics of products of different origins, but also for quality control and regulatory purposes. To date, official monographs or official analyses that allow researchers to evaluate propolis in a proper way have not yet been released. This study focuses on the characterization of twenty-seven Italian propolis samples and the identification of chemical markers that define its geographical provenance. Total polyphenol (TP) and total flavonoid (TF) content, alongside the quantification of pinocembrin, chrysin, galangin, and caffeic acid phenethyl ester (CAPE), were identified as potential markers. Additionally, DPPH assays were conducted to evaluate the antiradical activity of propolis samples. Our findings demonstrated that TPs, TFs and pinocembrin differed in propolis of different origins, especially in samples from the islands. However, the quantification of the sum of chrysin and galangin and CAPE provided a clearer distinction of the geographical origin of the propolis samples. In contrast, the DPPH assay did not prove useful for this purpose, as most results were similar and, therefore, not significant. This study lays the groundwork for future research on propolis. These findings could contribute to the development of more refined methods for distinguishing propolis origins, enhancing the understanding, valuation and quality control of this natural product in various applications. [ABSTRACT FROM AUTHOR]

Spirulina (Arthrospira platensis) is a protein-rich source because it contains approximately 60% protein. Therefore, it has positive impacts on human health, including anti-diabetes. The potential of Spirulina platensis as an anti-diabetic agent has been investigated in-vitro and in-vivo. Its extract is capable of inhibiting amylase, α-glucosidase, and DPP IV enzymes in a range of medium activities. Furthermore, spirulina at 250 mg/Kg body weight (BW) in mice can reduce blood glucose levels with similar outcomes to the positive controls (pioglitazone and glibenclamide). Protein groups (Phycocyanin), isolated peptides, and flavonoid groups such as Pinocembrin, Acacetin, and others have been predicted to be responsible for lowering glucose levels. In the market, several commercial products have been sold. Many commercial products have been sold in the market. However, these products are food supplements because a clinical study has not yet been undertaken to confirm their potency and safety. According to this analysis, there is a significant chance to produce anti-diabetic medications from spirulina sources in the future. [ABSTRACT FROM AUTHOR]

During the last two decades, many p38α mitogen-activated protein kinase (p38α MAPK) inhibitors have been developed and tested in preclinical/clinical studies for the treatment of various disorders, especially problems with the origin of inflammation. Previous studies strongly suggest the involvement of the p38α MAPK pathway in the pathogenesis of neurodegenerative disorders. Despite the significant progress made in this field, so far no studies have focused on p38α MAPK inhibitors that have the capability to be used for the treatment of neurodegenerative disorders. In the present review, we evaluated a wide range of well-known p38α MAPK inhibitors (more than 140 small molecules) by measuring key physicochemical parameters to identify those capable of successfully crossing the blood-brain barrier (BBB). As a result, we identify about 50 naturally occurring and synthetic p38α MAPK inhibitors with high potential to cross the BBB, which can be further explored in the future for the treatment of neurodegenerative disorders. In addition, a detailed analysis of the previously released X-ray crystal structure of the inhibitors in the active site of the p38α MAPK enzyme revealed that some residues such as Met109 play a critical role in the occurrence of effective interactions by constructing strong H-bonds. This study can encourage scientists to focus more on the design, production, and biological evaluation of new central nervous system (CNS)-active p38α MAPK inhibitors in the future. [ABSTRACT FROM AUTHOR]

In Indonesia, Boesenbergia rotunda, also referred to as Temu kunci. According to earlier research, B. rotunda is a rich source of flavonoids and other phenolic compounds. Extensive research is required to examine the chemical profiles of indigenous plants and analyze medicinal properties from extracts of B. rotunda. Thus, a phytochemical investigation is conducted on the extracts of B. rotunda and two recognized flavonoids, pinostrobin (1) and pinocembrin (2), were isolated from extracts of B. rotunda utilizing a variety of chromatographic methods. The structures are determined using the FTIR, and HPLC, 1H and 13C NMR data and comparison with published literature. The Physicochemical, pharmacokinetic, and toxicity prediction of substances (pKCSM) online tool was also used in this work to estimate the pharmacokinetic and toxicity potencies of pinostrobin and pinocembrin. The pKCSM prediction indicates that pinocembrin and pinostrobin have good pharmacokinetic characteristics and lack the potential to be hepatotoxic or mutagenic substances. Both pinostrobin and pinocembrin have low level lethal dosage values (LD50) of 2000 mol/kg. However, more investigation is still required to validate these hypotheses.

Background/objective: As the pivotal cellular mediators of bone resorption and pathological bone remodeling, osteoclasts have emerged as a prominent target for anti-resorptive interventions. Pinocembrin (PIN), a predominant flavonoid found in damiana, honey, fingerroot, and propolis, has been recognized for its potential therapeutic effects in osteolysis. The purpose of our project is to investigate the potential of PIN to prevent bone resorption in ovariectomized (OVX) mice by suppressing osteoclast production through its underlying mechanisms. Methods: The study commenced by employing protein-ligand molecular docking to ascertain the specific interaction between PIN and nuclear factor-κB (NF-κB) ligand (RANKL). Subsequently, PIN was introduced to bone marrow macrophages (BMMs) under the stimulation of RANKL. The impact of PIN on osteoclastic activity was assessed through the utilization of a positive TRAcP staining kit and a hydroxyapatite resorption assay. Furthermore, the study investigated the generation of reactive oxygen species (ROS) in osteoclasts induced by RANKL using H2DCFDA. To delve deeper into the underlying mechanisms, molecular cascades triggered by RANKL, including NF-κB, ROS, calcium oscillations, and NFATc1-mediated signaling pathways, were explored using Luciferase gene report, western blot analysis, and quantitative real-time polymerase chain reaction. Moreover, an estrogen-deficient osteoporosis murine model was established to evaluate the therapeutic effects of PIN in vivo. Results: In this study, we elucidated the profound inhibitory effects of PIN on osteoclastogenesis and bone resorption, achieved through repression of NF-κB and NFATc1-mediated signaling pathways. Notably, PIN also exhibited potent anti-oxidative properties by mitigating RANKL-induced ROS generation and augmenting activities of ROS-scavenging enzymes, ultimately leading to a reduction in intracellular ROS levels. Moreover, PIN effectively abrogated the expression of osteoclast-specific marker genes (Acp5, Cathepsin K, Atp6v0d2, Nfatc1, c-fos, and Mmp9), further underscoring its inhibitory impact on osteoclast differentiation and function. Additionally, employing an in vivo mouse model, we demonstrated that PIN effectively prevented osteoclast-induced bone loss resultant from estrogen deficiency. Conclusion: Our findings highlight the potent inhibitory effects of PIN on osteoclastogenesis, bone resorption, and RANKL-induced signaling pathways, thereby establishing PIN as a promising therapeutic candidate for the prevention and management of osteolytic bone diseases. The translational potential of this article: PIN serves as a promising therapeutic agent for the prevention and management of osteolytic bone diseases and holds promise for future clinical applications in addressing conditions characterized by excessive bone resorption. PIN is a natural compound found in various sources, including damiana, honey, fingerroot, and propolis. Its widespread availability and potential for therapeutic use make it an attractive candidate for further investigation and development as a clinical intervention.

In this study, the cytotoxic and apoptotic effects of three phenolic compounds highly found in the poplar type propolis; pinostrobin (PS), pinocembrin (PC) and pinobanksin (PB), were investigated individually and in combination on hormon-positive (MCF-7) and triple negative (MDA-MB-231) breast cancer cell lines and fibrocystic breast epithelium (MCF-10A) as control. Assessment of cytotoxicity and apoptosis were performed with WST-1 and Annexin V-7AAD assays, respectively. All statistical analyses including the two-way ANOVA and multiple-t-test were performed using GraphPad Prism software. Individually, PB (P0.05). On MCF-7, the triple combination induced cytotoxic/apoptotic effects even with the 25% dose and 50% dose on MDA-MB-231(p

A method for quantitative determination of pinocembrin in herb of licorice (Glycyrrhiza glabra L.) using high-performance liquid chromatography (HPLC) was developed. The content of the dominant flavonoid pinocembrin (5,7-dihydroxyflavanone) in G. glabra herb varied from 0.39 ± 0.007% to 1.28 ± 0.022%. The error of a single determination of the pinocembrin content in G. glabra herb was ±6.30% with confidence probability 95%. [ABSTRACT FROM AUTHOR]

In this study, the cytotoxic and apoptotic effects of three phenolic compounds highly found in the poplar type propolis; pinostrobin (PS), pinocembrin (PC) and pinobanksin (PB), were investigated individually and in combination on hormon-positive (MCF-7) and triple negative (MDA-MB-231) breast cancer cell lines and fibrocystic breast epithelium (MCF-10A) as control. Assessment of cytotoxicity and apoptosis were performed with WST-1 and Annexin V-7AAD assays, respectively. All statistical analyses including the two-way ANOVA and multiple-t-test were performed using GraphPad Prism software. Individually, PB (P<0.0001), PS (P<0.0001), and PC (P<0.05) demonstrated potent cytotoxic effects at moderate to high doses and late time intervals on MCF-7. PB and PS have been found to have a significant proliferative effect at low doses (P<0.0001), however, this effect disappeared in higher doses in this cell line. Dual combinations of PB+PC and PB+PS were toxic on MCF-10A, however, dual combination of PS+PC and the triple combination (PB+PS+PC) showed no cytotoxicity until high doses at late time intervals (P>0.05). On MCF-7, the triple combination induced cytotoxic/apoptotic effects even with the 25% dose and 50% dose on MDA-MB-231(p<0.0001). Our findings clearly showed that different combinations of these phenolic substances can have synergistic cytotoxic effects and even hormetic effects in non-tumorogenic cells. [ABSTRACT FROM AUTHOR]

Background: Hepatocellular carcinoma (HCC) is the most prevalent primary liver cancer. Anomianthus dulcis (Dunal) J.Sinclair (syn. Uvaria dulcis) has been used in Thai traditional medicine in various therapeutic indications. Phytochemical constituents of A. dulcis have been isolated and identified. However, their effects on liver cancer and the associated mechanisms have not been elucidated. Methods: Dry flowers of A. dulcis were extracted using organic solvents, and chromatographic methods were used to purify the secondary metabolites. The chemical structures of the pure compounds were elucidated by analysis of spectroscopic data. Cytotoxicity against HCC cells was examined using SRB assay, and the effects on cell proliferation were determined using flow cytometry. The mechanisms underlying HCC inhibition were examined by molecular docking and verified by Western blot analysis. Results: Among 3 purified flavonoids, pinocembrin, pinostrobin, and chrysin, and 1 indole alkaloid (3-farnesylindole), only pinocembrin showed inhibitory effects on the proliferation of 2 HCC cell lines, HepG2 and Li-7, whereas chrysin showed specific toxicity to HepG2. Pinocembrin was then selected for further study. Flow cytometric analyses revealed that pinocembrin arrested the HCC cell cycle at the G1 phase with a minimal effect on cell death induction. Pinocembrin exerted the suppression of STAT3, as shown by the molecular docking on STAT3 with a better binding affinity than stattic, a known STAT3 inhibitor. Pinocembrin also suppressed STAT3 phosphorylation at both Tyr705 and Ser727. Cell cycle regulatory proteins under the modulation of STAT3, namely cyclin D1, cyclin E, CDK4, and CDK6, are substantially suppressed in their expression levels. Conclusion: Pinocembrin extracted from A. dulcis exerted a significant growth inhibition on HCC cells via suppressing STAT3 signaling pathways and its downstream-regulated genes. [ABSTRACT FROM AUTHOR]

Licorice herb (Glycyrrhiza glabra L.) is a promising herbal raw material, which can be comprehensively used to develop drugs with an anti-inflammatory action.The aim of the article was to development a quantitative determination method of total flavonoids in Glycyrrhiza glabra L. herbs.Materials and methods. The subjects of research were 5 samples of licorice herb harvested in summer in various places of growing and cultivation. Pinostrobin was used as a standard sample. The registration of the electronic spectra was carried out with a spectrophotometer (Analytik Jena AG, Germany) by differential spectrophotometry, 96% ethanol was used as a solvent.Results. The methods for quantitative determination of total flavonoids in Glycyrrhiza glabra L. was carried out at an analytical wavelength of 310 nm equivalent to pinocembrin. The optimum parameters for the extraction of total flavonoids from Glycyrrhiza glabra L. were as follows: the extractant – 90% ethanol; the «raw material-extractant» ratio was 1:50; the extraction time was 60 min; the degree of atomization was 2 mm. The content of total flavonoids for the Glyccyrhiza glabra L. herb has been determined, it varies from 0.39±0.002 to 3.41±0.015% with the humidity of the vegetative raw material from 9.97±0.003 to 10.03±0.003% depending on the place of the vegetation, cultivation and year of the raw material collection. The error of the single determination with a 95% confidence level was ±0.73%.Conclusion. The developed methods for the quantitative determination of total flavonoids in Glycyrrhiza glabra L. herbs can be used to solve the issues of standardization of these medicinal plant raw materials.

Knowledge of the chemical composition of propolis is crucial for understanding the characteristics of products of different origins, but also for quality control and regulatory purposes. To date, official monographs or official analyses that allow researchers to evaluate propolis in a proper way have not yet been released. This study focuses on the characterization of twenty-seven Italian propolis samples and the identification of chemical markers that define its geographical provenance. Total polyphenol (TP) and total flavonoid (TF) content, alongside the quantification of pinocembrin, chrysin, galangin, and caffeic acid phenethyl ester (CAPE), were identified as potential markers. Additionally, DPPH assays were conducted to evaluate the antiradical activity of propolis samples. Our findings demonstrated that TPs, TFs and pinocembrin differed in propolis of different origins, especially in samples from the islands. However, the quantification of the sum of chrysin and galangin and CAPE provided a clearer distinction of the geographical origin of the propolis samples. In contrast, the DPPH assay did not prove useful for this purpose, as most results were similar and, therefore, not significant. This study lays the groundwork for future research on propolis. These findings could contribute to the development of more refined methods for distinguishing propolis origins, enhancing the understanding, valuation and quality control of this natural product in various applications.

Năm hợp chất gồm hai dihydrochalcone, 2’,6’-dihydroxy-4’-methoxydihydrochalcone (1), dihydroflavokawin B (2), một flavanone, pinocembrin (3) và hai diarylheptanoid, dihydroyashabushiketol (4), 5-methoxy-1,7-diphenyl-3-heptanone (5) đã được phân lập lần đầu tiên từ cao chiết n-hexane của loài Conamomum rubidum. Cấu trúc hóa học của chúng được xác định bằng phương pháp phổ cộng hưởng từ hạt nhân và so sánh với các dữ kiện phổ đã công bố. Hoạt tính kháng viêm trên mô hình ức chế sản sinh NO và gây độc tế bào của các hợp chất đã được đánh giá. Theo đó, hợp chất 2, 4 và 5 ức chế sản sinh NO với các giá trị IC50 trong khoảng từ 58,16 ± 3,02 đến 81,95 ± 2,87 µM. Hợp chất 1 ức chế tế bào ung thư phổi SK-LU-1 với giá trị IC50 = 58,45 ± 2,26 µM.

Anthrax toxin released by virulent strains of the bacterium, Bacillus anthracis, plays key factor in anthrax disease. The main purpose of this study was to investigate the interaction between four flavonoid ligands including Rhamnetin, Apigenin, Tectochrysin, Pinocembrin and protective antigens. Bioinformatics checking was done by means of Molegro virtual docker and Chimera 1.7. Also, in order for more accuracy, servers like Swiss Dock and BSP-SLIM, and all outputs obtained from this software were compared with each other. The results demonstrated that Apigenin interacted with the Glu117 which is crucial part of binding to its ligand with -12.3453 kca/mol. Also, the highest Fullfitness among these four ligands attributed to Rhamnetin with -994.80 kcal/mol and the ΔG =-7.06 kcal/mol. Results demonstrated that every four ligands possessed interaction with protective antigen and so have inhibitory effect on its interaction with cell membranes but the inhibitory activity of Apigenin and Rhamnetin in interaction is stronger than others flavonoids. Results shown above bring up laboratory studies based on these flavonoids in order to produce an efficacious drug against anthrax. [ABSTRACT FROM AUTHOR]

Background. Litsea glaucencens Kuth is an aromatic plant used for food seasoning food and in Mexican traditional medicine. Among, L. glaucencens leaves properties, it has proven antibacterial activity which can be used against opportunistic pathogens like Listeria monocytogenes, a foodborne bacteria that is the causal agent of listeriosis, a disease that can be fatal in susceptible individuals. The aim of this work was to investigate the antibacterial activity of L. glaucescens Kuth leaf extracts against L. monocytogenes and to identify its bioactive components. Material and Methods. L. glaucences leaves were macerated with four solvents of different polarity (n-hexane, dichloromethane, ethyl acetate, and methanol). To determine the capacity to inhibit bacterial proliferation in vitro, agar diffusion and microdilution methods were used. Next, we determined the minimal bactericidal concentration (MBC). Finally, we determined the ratio of MBC/MIC. Metabolites present in the active methanolic extract from L. glaucescens Kuth (LgMeOH) were purified by normalphase open column chromatography. The structure of the antibacterial metabolite was determined using nuclear magnetic resonance (¹H, 13C, COSY, HSQC) and by comparison with known compounds. Results. The LgMeOH extract was used to purify the compound responsible for the observed antimicrobial activity. This compound was identified as 5,7-dihydroxyflavanone (pinocembrin) by analysis of its spectroscopic data and comparison with those described. The MIC and MBC values obtained for pinocembrin were 0.68 mg/mL, and the ratio MBC/MIC for both LgMeOH and pinocembrin was one, which indicates bactericidal activity. Conclusion. L. glaucences Kuth leaves and its metabolite pinocembrin can be used to treat listeriosis due the bactericidal activity against L. monocytogenes. [ABSTRACT FROM AUTHOR]

Background Litsea glaucencens Kuth is an aromatic plant used for food seasoning food and in Mexican traditional medicine. Among, L. glaucencens leaves properties, it has proven antibacterial activity which can be used against opportunistic pathogens like Listeria monocytogenes, a foodborne bacteria that is the causal agent of listeriosis, a disease that can be fatal in susceptible individuals. The aim of this work was to investigate the antibacterial activity of L. glaucescens Kuth leaf extracts against L. monocytogenes and to identify its bioactive components. Material and Methods L. glaucences leaves were macerated with four solvents of different polarity (n-hexane, dichloromethane, ethyl acetate, and methanol). To determine the capacity to inhibit bacterial proliferation in vitro, agar diffusion and microdilution methods were used. Next, we determined the minimal bactericidal concentration (MBC). Finally, we determined the ratio of MBC/MIC. Metabolites present in the active methanolic extract from L. glaucescens Kuth (LgMeOH) were purified by normal-phase open column chromatography. The structure of the antibacterial metabolite was determined using nuclear magnetic resonance (1H, 13C, COSY, HSQC) and by comparison with known compounds. Results The LgMeOH extract was used to purify the compound responsible for the observed antimicrobial activity. This compound was identified as 5,7-dihydroxyflavanone (pinocembrin) by analysis of its spectroscopic data and comparison with those described. The MIC and MBC values obtained for pinocembrin were 0.68 mg/mL, and the ratio MBC/MIC for both LgMeOH and pinocembrin was one, which indicates bactericidal activity. Conclusion L. glaucences Kuth leaves and its metabolite pinocembrin can be used to treat listeriosis due the bactericidal activity against L. monocytogenes.

Hepatocyte lipid accumulation in type 2 diabetes mellitus (T2DM) promotes the development of metabolic dysfunction-associated steatosis liver disease (MASLD), whereas no specific drug treatment is available. Pinocembrin assists in the treatment of many diseases, however, little is known about its protective effect on hepatic lipid metabolism disorders induced by T2DM. In this study, the ameliorative effects of pinocembrin on T2DM-induced hepatocyte lipid accumulation were investigated by in vitro experiments combined with a comprehensive analysis strategy of network pharmacology and molecular docking. High glucose and free fatty acid co-cultured HepG2 cell line was used. The results indicated that pinocembrin reduced lipid deposition by modulating the release of Cyclooxygenase-2 (COX-2)-mediated inflammatory factors and improving insulin resistance (IR) in HepG2 cells. Overall, the findings of this study suggested that pinocembrin ameliorated lipid accumulation and IR in vitro, and thus may be a potential therapeutic agent for the treatment of T2DM-induced hepatic steatosis.

Microbial biofactories allow the upscaled production of high-value compounds in biotechnological processes. This is particularly advantageous for compounds like flavonoids that promote better health through their antioxidant, anti-bacterial, anti-cancer and other beneficial effects but are produced in small quantities in their natural plant-based hosts. Bacteria like E. coli have been genetically modified with enzyme cascades to produce flavonoids like naringenin and pinocembrin from coumaric or cinnamic acid. Despite advancements in yield optimization, the production of these compounds still involves high costs associated with their biosynthesis, purification, storage and transport. An alternative production strategy could involve the direct delivery of the microbial biofactories to the body. In such a strategy, ensuring biocontainment of the engineered microbes in the body and controlling production rates are major challenges. In this study, these two aspects are addressed by developing engineered living materials (ELMs) consisting of probiotic microbial biofactories encapsulated in biocompatible hydrogels. Engineered probiotic E. coli Nissle 1917 able to efficiently convert cinnamic acid into pinocembrin were encapsulated in poly(vinyl alcohol)-based hydrogels. The biofactories are contained in the hydrogels for a month and remain metabolically active during this time. Control over production levels is achieved by the containment inside the material, which regulates bacteria growth, and by the amount of cinnamic acid in the medium.

P38 mitogen‐activated protein kinase (p38 MAPK) signaling pathway is closely related to severe acute respiratory syndrome coronavirus‐2 (SARS‐CoV‐2) replication and hyperinflammatory responses in coronavirus disease 2019 (COVID‐19). Therefore, blood–brain barrier‐penetrating p38 MAPK inhibitors have good potential for the treatment of central nervous system (CNS) complications of COVID‐19. The aim of the present study is the characterization of the therapeutic potential of tanshinone IIA and pinocembrin for the treatment of CNS complications of COVID‐19. Studies published in high‐quality journals indexed in databases Scopus, Web of Science, PubMed, and so forth were used to review the therapeutic capabilities of selected compounds. In continuation of our previous efforts to identify agents with favorable activity/toxicity profiles for the treatment of COVID‐19, tanshinone IIA and pinocembrin were identified with a high ability to penetrate the CNS. Considering the nature of the study, no specific time frame was determined for the selection of studies, but the focus was strongly on studies published after the emergence of COVID‐19. By describing the association of COVID‐19‐induced CNS disorders with p38 MAPK pathway disruption, this study concludes that tanshinone IIA and pinocembrin have great potential for better treatment of these complications. The inclusion of these compounds in the drug regimen of COVID‐19 patients requires confirmation of their effectiveness through the conduction of high‐quality clinical trials. [ABSTRACT FROM AUTHOR]

Random skin flap grafting is the most common skin grafting technique in reconstructive surgery. Despite progress in techniques, the incidence of distal flap necrosis still exceeds 3%, which limits its use in clinical practice. Current methods for treating distal flap necrosis are still lacking. Pinocembrin (Pino) can inhibit reactive oxygen species (ROS) and cell death in a variety of diseases, such as cardiovascular diseases, but the role of Pino in random flaps has not been explored. Therefore, we explore how Pino can enhance flap survival and its specific upstream mechanisms via macroscopic examination, Doppler, immunohistochemistry, and western blot. The results suggested that Pino can enhance the viability of random flaps by inhibiting ROS, pyroptosis and apoptosis. The above effects were reversed by co‐administration of Pino with adeno‐associated virus‐silencing information regulator 2 homolog 3 (SIRT3) shRNA, proving the beneficial effect of Pino on the flaps relied on SIRT3. In addition, we also found that Pino up‐regulates SIRT3 expression by activating the AMP‐activated protein kinase (AMPK) pathway. This study proved that Pino can improve random flap viability by eliminating ROS, and ROS‐induced cell death through the activation of SIRT3, which are triggered by the AMPK/PGC‐1α signaling pathway. [ABSTRACT FROM AUTHOR]

Re-epithelialization is delayed in aged skin due to a slow rate of keratinocyte proliferation, and this may cause complications. Thus, there has been development of new therapies that increase treatment efficacy for skin wounds. Epidermal growth factor (EGF) has been clinically used, but this agent is expensive, and its activity is less stable. Therefore, a stable compound possessing EGF-like properties may be an effective therapy, especially when combined with EGF. The current study discovered that pinocembrin (PC) effectively synergized with EGF in increasing keratinocyte viability. The combination of PC and EGF significantly enhanced the proliferation and wound closure rate of the keratinocyte monolayer through activating the phosphorylation of ERK and Akt. Although these effects of PC were like those of EGF, we clearly proved that PC did not transactivate EGFR. Recent data from a previous study revealed that PC activates G-protein-coupled receptor 120 which further activates ERK1/2 and Akt phosphorylation. Therefore, this clearly indicates that PC possesses a unique property to stimulate the growth and survival of keratinocytes through activating a different receptor, which subsequently conveys the signal to cross-talk with the effector kinases downstream of the EGFR, suggesting that PC is a potential compound to be combined with EGF. [ABSTRACT FROM AUTHOR]

Objective To explore the effect and mechanism of pinocembrin on the proliferation, migration and invasion of gastric cancer cell line AGS. Methods AGS cells were divided into different test groups according to concentrations of pinocembrin(50, 100, 200, 400 μmol/L), NC group, 200 μmol/L pinocembrin + anti-miR-NC group, 200 μmol/L pinocembrin + anti-miR-34a-5p group. The proliferation, apoptosis, migration, invasion ability and protein expression of AGS cells were detected by CCK-8 assay, flow cytometry, Transwell chamber assay and Western blot. The expression of miR-34a-5p was detected by real-time fluorescent quantitative PCR (RT-qPCR). Results Different concentrations of pinocembrin reduced the proliferation activity of AGS cells (P＜0.05) and the proportion of in G0/G1 phase cells was increased while the proportion of S phase cells decreased. The expressions of MMP2, MMP9, p-PI3K and p-AKT protein decreased. The cell migration and invasion were inhibited. The expression of miR-34a-5p was increased (P＜0.05), which reduced the inhibitory effect of pinocembrin on the prolifera- tion, migration and invasion of AGS cells. Conclusions Pinocembrin can inhibit the proliferation, migration and invasion of AGS cells by improving the expression of miR-34a-5p.

Acrolein (ACR) is a toxic unsaturated aldehyde that is produced during food thermal processing. Here, we investigated the synergistic effect of polyphenols in binary, ternary, and quaternary combinations on ACR by the Chou–Talalay method, and then explored the synergistic effect of cardamonin (CAR), alpinetin (ALP), and pinocembrin (PIN) in fixed proportion from Alpinia katsumadai Hayata (AKH) combined with curcumin (CUR) in the model, and roasted pork using LC–MS/MS. Our results showed that their synergistic effect depended on the intensification of their individual trapping ACR activities, which resulted in the formation of more ACR adducts. In addition, by adding 1% AKH (as the carrier of CAR, ALP, and PIN) and 0.01% CUR (vs. 6% AKH single) as spices, more than 71.5% (vs. 54.0%) of ACR was eliminated in roast pork. Our results suggested that selective complex polyphenols can synergistically remove the toxic ACR that is produced in food processing. [ABSTRACT FROM AUTHOR]

• Pinocembrin is identified as a potential anti-obesity agent using CMap. • PB promotes adipose tissue lipolysis and inhibit lipid formation to anti-obesity. • PB inhibits the formation of lipid droplets in preadipocytes through GPR120-ERK1/2 signaling pathway. Obesity is a widely concerned health problem. Mobilizing white adipose tissue and reducing fat synthesis are considered as effective strategies in the treatment of obesity. Here, using Connectivity Map (CMap) approach, we identified the pinocembrin (PB), a natural flavonoid primarily found in propolis, as a potential anti-obesity drug. Therefore, high-fat-diet (HFD) mice were randomly divided into two groups and fed a HFD or HFD with PB in this study. In vivo experiments showed that supplementation of PB reduced the body weight gain and ameliorated insulin resistance in HFD-induced mice. More importantly, PB did not cause side effect through detecting the levels of alanine transaminase (ALT), aspartate aminotransferase (AST), creatinine (CRE) and blood urea nitrogen (BUN) in serum of mice. Additionally, PB reduced expansion of white adipose tissue with upregulation of genes related lipolysis and downregulation of genes related lipogenesis. Furthermore, in vitro experiments revealed that PB treatment dose-dependently inhibited lipid droplet formation with upregulation of genes related lipolysis and downregulation of genes related lipogenesis. Molecular docking analysis combined with cellular thermal shift assay (CETSA) suggested that PB has a high affinity to the G protein-coupled receptor 120 (GPR120). Meanwhile, we confirmed that PB efficiently inhibited adipogenic differentiation of preadipocytes by directly binding to GPR120 and subsequently activating the downstream phosphorylation extracellular regulated kinase 1/2 (ERK1/2). Collectively, PB exerted anti-obesity effect through GPR120-ERK1/2 signaling pathway, providing a novel and promising natural drug for the treatment of obesity. [ABSTRACT FROM AUTHOR]

Abstract Background Malaria remains the major health problem responsible for many mortality and morbidity in developing countries. Because of the development of resistance by Plasmodium species, searching effective antimalarial agents becomes increasingly important. Pinocembrin is a flavanone previously isolated as the most active antiplasmodial compound from the leaves of Dodonaea angustifolia. For a better understanding of the antiplasmodial activity, the synthesis of pinocembrin and a great number of analogs was undertaken. Methods Chalcones 5a-r were synthesized via Claisen-Schmidt condensation using 2,4-dibenzyloxy-6-hydroxyacetophenone and aromatic aldehydes as substrates under basic conditions. Cyclization of compounds 5a-r to the corresponding dibenzylated pinocembrin analogs 6a-r was achieved using NaOAc in EtOH under reflux. Catalytic hydrogenation using 10% Pd/C as catalyst in an H-Cube Pro was used for debenzylation to deliver 7a-l. The structures of the synthesized compounds were characterized using various physical and spectroscopic methods, including mp, UV, IR, NMR, MS and HRMS. The synthesized dibenzylated flavanones 6a-d, 6i and 7a were evaluated for their in vivo antiplasmodial activities against Plasmodium berghei infected mice. Molecular docking simulation and drug likeness properties of compounds 7a-l were assessed using AutoDock Vina and SwissADME, respectively. Results A series of chalcones 5a-r has been synthesized in yields ranging from 46 to 98%. Treatment of the chalcones 5a-r with NaOAc refluxing in EtOH afforded the dibenzylated pinocembrin analogs 6a-r with yields up to 54%. Deprotection of the dibenzylated pinocembrin analogs delivered the products 7a-l in yields ranging from 78 to 94%. The dibenzylated analogs of pinocembrin displayed percent inhibition of parastaemia in the range between 17.4 and 87.2% at 30 mg/kg body weight. The parastaemia inhibition of 87.2 and 55.6% was obtained on treatment of the infected mice with pinocembrin (7a) and 4’-chloro-5,7-dibenzylpinocembrin (6e), respectively. The mean survival times of those infected mice treated with these two compounds were beyond 14 days indicating that the samples suppressed P. berghei and reduced the overall pathogenic effect of the parasite. The molecular docking analysis of the chloro derivatives of pinocembrin revealed that compounds 7a-l show docking affinities ranging from – 8.1 to – 8.4 kcal/mol while it was -7.2 kcal/mol for chloroquine. Conclusion Pinocembrin (7a) and 4’-chloro-5,7-dibenzyloxyflavanone (6e) displayed good antiplasmodial activity. The in silico docking simulation against P. falciparum dihydrofolate reductase-thymidylate synthase revealed that pinocembrin (7a) and its chloro analogs 7a-l showed better binding affinity compared with chloroquine that was used as a standard drug. This is in agreement with the drug-like properties of compounds 7a-l which fulfill Lipinski's rule of five with zero violations. Therefore, pinocembrin and its chloro analogs could serve as lead compounds for further antiplasmodial drug development.

Some of the challenges to developing greener sample preparation procedures are related to finding solvents and practices with low environmental impacts. Given the importance of CO2, water, and ethanol compared to other green solvents in flavonoid extractions, it is convenient to explore changes in how they are employed. Depending on temperature and pressure, these three solvents may present total or partial miscibility that can be used conveniently in sample preparation. In this work, the Lippia origanoides (Verbenaceae family) vegetal material remnant after essential oil distillation was extracted with either aqueous ethanol (EtOH), ethanol-modified supercritical CO2 (EtOHCO2), or two coexisting CO2 fluid phases [(CO2)2]. The latter was the extractive practice that afforded higher selectivity and yield of pinocembrin (Pn) and galangin (Gn), two important active ingredients for pharmaceutical applications. EtOH extraction was the practice with the highest whole yield, and its extract contained mainly glycosylated compounds, in contrast to those extraction systems that involved CO2. The presence of CO2 allowed selective extraction of nonglycosylated flavonoids, possibly due to π−π intermolecular interactions with them. Flavonoids whose B-ring is a benzene or phenol group were recovered in higher amount. By means of the EtOHCO2 and (CO2)2 techniques, the extraction/isolation of Pn and Gn was achieved with less ethanol consumption and lower environmental impacts. The best setup was extraction with (CO2)2 and isolation by preparative HPLC. These results are promising for increasing selectivity and yield in some specific sample preparations.

Stroke is one of the main causes of mortality and disability, and it is due to be included in monetary implications on wellbeing frameworks around the world. Ischemic stroke is caused by interference in cerebral blood flow, leading to a deficit in the supply of oxygen to the affected region. It accounts for nearly 80–85% of all cases of stroke. Oxidative stress has a significant impact on the pathophysiologic cascade in brain damage leading to stroke. In the acute phase, oxidative stress mediates severe toxicity, and it initiates and contributes to late-stage apoptosis and inflammation. Oxidative stress conditions occur when the antioxidant defense in the body is unable to counteract the production and aggregation of reactive oxygen species (ROS). The previous literature has shown that phytochemicals and other natural products not only scavenge oxygen free radicals but also improve the expressions of cellular antioxidant enzymes and molecules. Consequently, these products protect against ROS-mediated cellular injury. This review aims to give an overview of the most relevant data reported in the literature on polyphenolic compounds, namely, gallic acid, resveratrol, quercetin, kaempferol, mangiferin, epigallocatechin, and pinocembrin, in terms of their antioxidant effects and potential protective activity against ischemic stroke. [ABSTRACT FROM AUTHOR]

Objective　 To observe the preventive effect of pinocembrin (PIN) on myocardial cell death induced by hypoxia/reoxygenation (H/R) in rats, and to explore its preventive mechanism. Methods　 The rat cardiomyocytes H9c2 were divided into PIN-H+740-Y-P group, PIN-H group, PIN-L group, model group, and control group. H9c2 cells in the PIN-H+740-Y-P group were incubated with medium including 25 μmol/L PIN and 50 μg/mL 740-Y-P, PIN-H group with medium including 25 μmol/L PIN, PIN-L group with medium including 12. 5 μmol/L PIN and model group with blank medium for 1 h and then followed by H/R treatment. H9c2 cells in the control group were only cultured with blank medium. The cells of the above groups were cultured for 24, 48 and 72 h, and the cell viability was determined by CCK-8. We took the cells of the above groups, measured the proportion of myocardial cells in each group by pyridine io⁃ dide (PI) staining, and measured the LDH in the cells by ELISA, and detected the phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT) and nuclear factor- κB (NF- κB) protein, and pyrolytic related proteins ［Caspase-1, apoptosis-associated speck-like protein containing CARD (ASC) and NOD-like receptor protein 3 (NLRP3) proteins］ in the cells by Western blotting. PI3K, AKT and NF-κB mRNA in cells were determined by fluorescence quantitative PCR. Results　 Compared with the control group, the cell viability (at 48 and 72 h) decreased (P<0. 05), but the cardiomyo⁃ cyte pyroptosis ratio, LDH level, PI3K, Akt and NF-κB mRNA expression levels, PI3K, Akt, NF-κB p65 protein phosphorylation levels, Caspase-1, ASC and NLRP3 protein expression levels increased in the H/R group (all P<0. 05). Com⁃ pared with the model group, the cell viability increased (at 48 and 72 h) (P<0. 05), but the proportion of myocardial cell death, the level of LDH, the relative expression levels of Caspase-1, ASC and NLRP3 proteins, the phosphorylation levels of PI3K, Akt and NF-κB p65 proteins and the relative expression levels of PI3K, Akt and NF-κB mRNA decreased in the PIN-H group (all P<0. 05). Compared with the PIN-H group, the cell viability decreased, but the proportion of myocardial cell death, the level of LDH, the relative expression levels of Caspase-1, ASC, NLRP3 proteins, the phosphorylation levels of PI3K, AKT, NF-κB p65 protein and the relative mRNA expression levels of PI3K, AKT and NF-κB increased in the PIN-H+740-Y-P group (all P<0. 05). Conclusion　 PIN may inhibit H/R-induced cardiomyocyte pyroptosis by downregulating PI3K/AKT/NF-κB signaling pathway. [ABSTRACT FROM AUTHOR]

Hepatic ischemia-reperfusion injury (HIRI) is of common occurrence during liver surgery and transplantation. Pinocembrin (PIN) is a kind of flavonoid monomer extracted from the local traditional Chinese medicine Penthorum chinense Pursh (P. chinense). However, the effect of PIN on HIRI has not determined. We investigated the protective effect and potential mechanism of PIN against HIRI. Model mice were subjected to partial liver ischemia for 60 min, experimental mice were pretreated with PIN orally for 7 days, and H2O2-induced oxidative damage model in AML12 hepatic cells was established in vitro. Histopathologic analysis and serum biochemical levels revealed that PIN had hepatoprotective activities against HIRI. The variation of GSH, SOD, MDA, and ROS levels indicated that PIN treatments attenuated oxidative stress in tissue. PIN pretreatment obviously ameliorated apoptosis, and restrained the expression of HMGB1 and TLR4 in vivo. In vitro, compared with H2O2 group, the contents of ROS, mitochondrial membrane potential, apoptotic cells, and Bcl-2 protein were decreased, while the Bax protein expression was increased. Moreover, HMGB-1 small interfering RNA test and western blotting showed that PIN pretreatment reduced HMGB1 and TLR4 protein levels. In conclusion, PIN pretreatment effectively protected hepatocytes from HIRI and inhibited the HMGB1/TLR4 signaling pathway. [ABSTRACT FROM AUTHOR]

Background: Propolis is a natural resinous mixture produced by bees. It provides beneficial effects on human health in the treatment/management of many diseases. The present study was performed to demonstrate the anti-Acanthamoeba activity of ethanolic extracts of Propolis samples from Iran. The interactions of the compounds and essential proteins of Acanthamoeba were also visualized through docking simulation. Methods: The minimal inhibitory concentrations (MICs) of Propolis extract against Acanthamoeba trophozoites and cysts was determined in vitro. In addition, two-fold dilutions of each of agents were tested for encystment, excystment and adhesion inhibitions. Three major compounds of Propolis extract such as chrysin, tectochrysin and pinocembrin have been selected in molecular docking approach to predict the compounds that might be responsible for encystment, excystment and adhesion inhibitions of A. castellanii. Furthermore, to confirm the docking results, molecular dynamics (MD) simulations were also carried out for the most promising two ligand-pocket complexes from docking studies. Results: The minimal inhibitory concentrations (MICs) 62.5 and 125 µg/mL of the most active Propolis extract were assessed in trophozoites stage of Acanthamoeba castellanii ATCC30010 and ATCC50739, respectively. At concentrations lower than their MICs values (1/16 MIC), Propolis extract revealed inhibition of encystation. However, at 1/2 MIC, it showed a potential inhibition of excystation and anti-adhesion. The molecular docking and dynamic simulation revealed the potential capability of Pinocembrin to form hydrogen bonds with A. castellanii Sir2 family protein (AcSir2), an encystation protein of high relevance for this process in Acanthamoeba. Conclusions: The results provided a candidate for the development of therapeutic drugs against Acanthamoeba infection. In vivo experiments and clinical trials are necessary to support this claim.

Background: Propolis is a natural resinous mixture produced by bees. It provides beneficial effects on human health in the treatment/management of many diseases. The present study was performed to demonstrate the anti-Acanthamoeba activity of ethanolic extracts of Propolis samples from Iran. The interactions of the compounds and essential proteins of Acanthamoeba were also visualized through docking simulation. Methods: The minimal inhibitory concentrations (MICs) of Propolis extract against Acanthamoeba trophozoites and cysts was determined in vitro. In addition, two-fold dilutions of each of agents were tested for encystment, excystment and adhesion inhibitions. Three major compounds of Propolis extract such as chrysin, tectochrysin and pinocembrin have been selected in molecular docking approach to predict the compounds that might be responsible for encystment, excystment and adhesion inhibitions of A. castellanii. Furthermore, to confirm the docking results, molecular dynamics (MD) simulations were also carried out for the most promising two ligand-pocket complexes from docking studies. Results: The minimal inhibitory concentrations (MICs) 62.5 and 125 µg/mL of the most active Propolis extract were assessed in trophozoites stage of Acanthamoeba castellanii ATCC30010 and ATCC50739, respectively. At concentrations lower than their MICs values (1/16 MIC), Propolis extract revealed inhibition of encystation. However, at 1/2 MIC, it showed a potential inhibition of excystation and anti-adhesion. The molecular docking and dynamic simulation revealed the potential capability of Pinocembrin to form hydrogen bonds with A. castellanii Sir2 family protein (AcSir2), an encystation protein of high relevance for this process in Acanthamoeba. Conclusions: The results provided a candidate for the development of therapeutic drugs against Acanthamoeba infection. In vivo experiments and clinical trials are necessary to support this claim.

Neuroinflammation is critical to the comorbidity of chronic pain and depression. Pyroptosis is an inflammatory cell death that is different from apoptosis. Activation of the P2X4 receptor leads to inflammation and is involved in chronic pain and depression. Pinocembrin (5,7-dihydroxyflavanone) is a natural flavonoid compound with anti-inflammatory, antioxidant and neuroprotective effects. In this study, an animal model of chronic pain and depression comorbidity was used to explore the therapeutic effect of pinocembrin in P2X4-mediated pyroptosis. The results showed that nociceptive behaviours and depression-like behaviours were obvious in the model rats induced by chronic constrictive injury (CCI) and chronic unpredictable mild stimulus (CUMS). In the model rats, the mRNA and protein levels of the P2X4 receptor in the hippocampus were increased, and the coexpression of P2X4 and the astrocyte marker glial fibrillary acidic protein (GFAP) in the hippocampus was increased. The protein content of connexion 43 (Cx43), NOD-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) and caspase-1 was increased. The serum content of IL-1β and the mRNA and protein expression of IL-1β were increased. The protein content of p-P38MAPK was increased. After treatment with pinocembrin in the model rats, these behavioural changes were improved, and the mRNA and protein levels of the above indicators were decreased. The results of molecular docking confirmed that the affinity of pinocembrin and the P2X4 receptor was − 7.8 (kcal/mol). At the same time, pinocembrin inhibited the ATP release and Ca2+ signal release in primary astrocytes and ATP-activated current of HEK293 cells transfected with the pcDNA3.0-EGFP-hP2X4 plasmid. Therefore, pinocembrin relieved nociceptive and depression-like behaviours in rats with chronic pain and depression comorbidity by inhibiting P2X4 receptor–mediated pyroptosis in the hippocampus. The mechanism of pinocembrin in treating rats with chronic pain and depression comorbidity. GJ stands for gap junction, and Cx43 is mainly expressed in astrocytes. [ABSTRACT FROM AUTHOR]

Pneumonia is a serious infectious disease with increased morbidity and mortality worldwide. The M. pneumoniae is a major airway pathogen that mainly affects respiratory tract and ultimately leads to the development of pneumonia. The current exploration was aimed to uncover the beneficial properties of pinocembrin against the M. pneumoniae-triggered pneumonia in mice via its anti-inflammatory property. The pneumonia was stimulated to the BALB/c mice via infecting them with M. pneumoniae (100 µl) for 2 days through nasal drops and concomitantly treated with pinocembrin (10 mg/kg) for 3 days. The azithromycin (100 mg/kg) was used as a standard drug. Then the lung weight, nitric oxide, and myeloperoxidase (MPO) activity was assessed. The content of MDA, GSH, and SOD activity was scrutinized using kits. The total cells and DNA amount present in the bronchoalveolar lavage fluid (BALF) was assessed by standard methods. The IL-1, IL-6, IL-8, TNF-α, and TGF contents in the BALF samples and NF-κB level in the lung tissues were assessed using kits. The lung histopathology was assessed microscopically to detect the histological alterations. The 10 mg/kg of pinocembrin treatment substantially decreased the lung weight, nitric oxide (NO) level, and MPO activity. The MDA level was decreased, and GSH content and SOD activity were improved by the pinocembrin treatment. The pinocembrin administered pneumonia animals also demonstrated the decreased total cells, DNA amount, IL-1, IL-6, IL-8, TNF-α, and TGF in the BALF and NF-κB level. The findings of histological studies also witnessed the beneficial role of pinocembrin against M. pneumoniae-infected pneumonia. In conclusion, our findings confirmed that the pinocembrin effectively ameliorated the M. pneumoniae-provoked inflammation and oxidative stress in the pneumonia mice model. Hence, it could be a hopeful therapeutic agent to treat the pneumonia in the future. [ABSTRACT FROM AUTHOR]

Pinocembrin is a type of flavonoid present in honey and propolis. It has high antioxidant and neuroprotective effect. The present study was designed to evaluate the immunomodulatory properties of pinocembrin (PCB) and the anti-apoptosis effects using in vitro and in vivo mice splenocytes and experimental autoimmune encephalomyelitis (EAE) in C57BL/6j female mice, respectively. The reduction of clinical symptoms and disease index were evaluated by histochemical analysis. T-cell population and cytokines levels in myelin oligodendrocyte glyco-protein (MOG) challenged PCB treated cells were estimated using flow cytometry and ELISA kits. Apoptotic and neuronal markers were evaluated using quantitative real time PCR and protein blots. Results showed that PCB inhibited the infiltration of inflammatory cells and improved the myelin protective proteins. It also positively regulated the antioxidants and apoptotic markers including Caspase-3, TGF-ß, SIRT-1, CCL-2 and MBP. Moreover, PCB modulated the levels of inflammatory mediators and TGF-ß in splenic cells. The production of myelin basic protein (MBP) increased in PCB treated EAE mice. Taken together, the current finding revealed that, PCB suppressed EAE pathological checkpoints for demyelination and apoptosis in CNS and act as a therapeutic role for treatment of MS in upcoming prospects. [ABSTRACT FROM AUTHOR]

To prepare polyethylene glycol succinate-vitamin E modified pinocembrin (PCB)-loaded liposomes (PCBT-liposomes) and evaluate PCBT-liposomal pharmacokinetics and antihyperglycemic activity. The novel PCBT-liposomes demonstrated a promising application prospect as a nano drug carrier for future research. Thin film dispersion was used to prepare PCBT-liposomes. We measured a series of characterization, followed by in vitro cumulative release, in vivo pharmacokinetic study, and antihyperglycemic activity evaluation. PCBT-liposomes displayed spherical and bilayered nanoparticles with mean particle size (roughly 92 nm), negative zeta potential (about −26.650 mV), high drug encapsulation efficiency (87.32 ± 1.34%) and good storage (at 4 or 25 °C) stability during 48 h after hydration. The cumulative release rate of PCBT-liposomes was markedly higher than free PCB in four different pH media. In vivo investigation showed that PCBT-liposomes could obviously improve oral bioavailability of PCB by 1.96 times, whereas the Cmax, MRT0–t, and T1/2 of PCBT-liposomes were roughly 1.700 ± 0.139 µg·mL−1, 12.695 ± 1.647 h, and 14.244 h, respectively. In terms of biochemical analysis, aspartate amino-transferase (AST), alanine amino-transferase (ALT), interleukin-1 (IL-1), and tumor necrosis factor-α (TNF-α) concentrations in serum of diabetic mice were respectively decreased 28.28%, 17.23%, 17.77%, and 8.08% after PCBT-liposomal treatment. These results show PCBT-liposomal preparation as an excellent nano-carrier which has the potential to improve water solubility, bioavailability, and antihyperglycemic activity of PCB, amid broadening the application of PCB in the clinical settings. [ABSTRACT FROM AUTHOR]

Plant-based food represents an excellent source of different nutrients and bioactive compounds, such as phenolics, carotenoids, vitamins, etc., with proven health benefits for humans. The content of selected phytochemicals, polyphenolic profile, and biological activity (antioxidant potential and α-glucosidase inhibitory activity) of fruit extracts of medlar (Mespilus germanica L.), blackthorn (Prunus spinosa L.), and common hawthorn (Crataegus monogyna Jacq.), the neglected Rosaceae species originated from Serbia were studied. Targeted UHPLC/(−)HESI–MS/MS quantitative analysis of phenolic compounds revealed pinocembrin only in medlar fruit extract, and it is the first report of this flavanone in medlar fruits. Total phenolic content did not differ between extracts, whereas significant differences were observed for the contents of total flavonoids, total phenolic acids, and total gallotannins. Monomeric anthocyanins and total anthocyanins were significantly higher in blackthorn compared to medlar and hawthorn fruit extracts (p < 0.05). DPPH· and ABTS·+ scavenging activities for examined fruits were modest compared to other natural antioxidants and BHT. The most potent inhibitory activity toward α-glucosidase expressed medlar and blackthorn extracts with IC50 values of 129.46 and 199.84 μg/mL, respectively, which was higher compared to the standard drug acarbose. [ABSTRACT FROM AUTHOR]

Background: Anxiety disorder (AD) is the most common mental disorder, which is closely related to atrial fibrillation (AF) and is considered to be a trigger of AF. Pinocembrin has been demonstrated to perform a variety of neurological and cardiac protective effects through its anti-inflammatory and antioxidant activities. The current research aims to explore the antiarrhythmic effect of pinocembrin in anxiety disorder rats and its underlying mechanisms. Methods: 60 male Sprague-Dawley rats were distributed into four groups: CTL group: control rats + saline; CTP group: control rats + pinocembrin; Anxiety disorder group: anxiety disorder rats + saline; ADP group: anxiety disorder rats + pinocembrin. Empty bottle stimulation was conducted to induce anxiety disorder in rats for 3 weeks, and pinocembrin was injected through the tail vein for the last 2 weeks. Behavioral measurements, in vitro electrophysiological studies, biochemical assays, ELISA, Western blot and histological studies were performed to assess the efficacy of pinocembrin. In addition, HL-1 atrial cells were cultured in vitro to further verify the potential mechanism of pinocembrin. Results: After 3 weeks of empty bottle stimulation, pinocembrin significantly improved the exploration behaviors in anxiety disorder rats. Pinocembrin alleviated electrophysiological remodeling in anxiety disorder rats, including shortening the action potential duration (APD), prolonging the effective refractory period (ERP), increasing the expression of Kv1.5, Kv4.2 and Kv4.3, decreasing the expression of Cav1.2, and ultimately reducing the AF susceptibility. These effects may be attributed to the amelioration of autonomic remodeling and structural remodeling by pinocembrin, as well as the inhibition of oxidative stress with upregulation of the nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) pathway. Conclusion: Pinocembrin can reduce AF susceptibility in anxiety disorder rats induced by empty bottle stimulation, with the inhibition of autonomic remodeling, structural remodeling, and oxidative stress. Therefore, pinocembrin is a promising treatment for AF in patients with anxiety disorder. [ABSTRACT FROM AUTHOR]

Acute kidney injury is a condition when kidney function decreased abruptly, characterized by e.g. a decrease in the glomerular filtration rate. This condition is due to the activation of various signaling pathways, such as effector caspases, and thus can reverse kidney damage by interacting with these proteins, and eventually inhibiting the apoptotic process. This study aims to determine the molecular interactions of eight phytoconstituents in B. rotunda rhizome with the active binding pocket of caspase-3 and-7 and to predict the ADMET profile of these ligands. Results indicated that all ligands could occupy the binding pocket of both caspases, similar to those of caspase inhibitors, which means that these compounds could inhibit the apoptotic signaling pathway, thus protecting the cell. However, these ligands indicate a better docking score with caspase-7 than with caspase-3. Of all ligands, 4-hydroxypanduratin A and panduratin A showed the best affinity (-8.3 kcal/mol and -8.5 kcal/mol, respectively). ADMET prediction revealed that the excellent oral bioavailability belongs to 4-hydroxypanduratin A and pinostrobin (HIA >99%; Caco2 >70 μm/sec; BBB <1; PPB <50%). [ABSTRACT FROM AUTHOR]

Chronic opioid induced analgesic tolerance is a major obstacle in pain management. Microglial activation is involved in morphine tolerance and pinocembrin suppresses microglial activation in several disease models. We aim to investigate whether and how pinocembrin alleviates morphine tolerance. We induced chronic morphine tolerance in mice by daily morphine injection, with some mice receiving pinocembrin. Analgesic tolerance and hyperalgesia were determined by behavioral assays. The effects of pinocembrin on morphine induced microglial activation, neuroinflammation, and STAT3 activation were determined by Iba1 immunostaining, Il1b and Tnfa mRNA levels and STAT3 phosphorylation. Finally, the effects of STAT3 inhibition on chronic morphine tolerance were assessed. We show that pinocembrin not only suppressed but also reversed preexisting chronic morphine tolerance and hyperalgesia. We found that chronic administration of morphine lead to microglial activation and neuroinflammation, which were suppressed by pinocembrin. Our results reveal a strong connection of STAT3 with morphine tolerance and pinocembrin suppressed morphine-induced STAT3 activation both in vivo and in BV2 cells. Finally, we show that STAT3 inhibition is sufficient to suppress morphine tolerance and hyperalgesia. Our study suggests that pinocembrin effectively prevents and alleviates chronic morphine tolerance through inhibition of STAT3 mediated microglia activation and neuroinflammation. [ABSTRACT FROM AUTHOR]

The main purpose of the present study was to evaluate the antitumor effects of pinocembrin in human prostate cancer cells (PC-3) along with investigating its effects on cell apoptosis, endogenous ROS production and cell cycle. MTT assay and clonogenic assays were used to study the effects on cell viability and cancer colony formation, respectively. Fluorescence microscopy along with Western blotting was used to study apoptotic effects induced by pinocembrin. Flow cytometry was used to study effects on ROS production and cell cycle phase distribution. Results indicated that pinocembrin promoted inhibition cell proliferation along with reducing cancer colony formation of PC-3 cells in a dose-dependent manner. Pinocembrin induced regulatory effects over expressions of caspase-3, caspase-9, Bax and Bcl-2, thereby promoting apoptotic cell death in PC-3 cells. It also led to the dose-dependent G0/G1 cell cycle arrest. In conclusion, pinocembrin exhibits strong anticancer effects in human prostate cancer cells mediated via apoptosis, endogenous ROS production and G0/G1 cell cycle arrest.

Wenqi Wang,&ast; Xin Feng,&ast; Yu Du, Cen Liu, Xinxin Pang, Kunxiu Jiang, Xirui Wang, Yonggang Liu School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, 102488, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yonggang Liu Email liuyg0228@163.comPurpose: Pinocembrin is a dihydroflavonoid, which is widely found in several plant species. Although pinocembrin has good pharmacological activity, it has poor water solubility and low bioavailability. Therefore, we have modified the structure of pinocembrin with a combination of different amino acids to solve this problem. Moreover, the effect of the antiaging activity of them has not been explored. We aim to investigate the effect of pinocembrin and its amino acid derivatives on the aging of Caenorhabditis elegans.Methods: Pinocembrin was spliced with different amino acids in order to obtain their corresponding derivatives. The preliminary research of pinocembrin and its amino acid derivatives on antiaging effect was studied by using the C. elegans model. Among all the compounds, the one shows the best antiaging effect was then studied on antiaging mechanism. The protective effect on nematodes under emergency conditions was explained by detecting the ROS content and sod-3p::GFP fusion protein expression in nematodes; the possible anti-aging mechanism of nematodes was determined by DAF-16 nuclear localization experiment and the survival curve of transgenic nematodes model under stress conditions.Results: Pb-3 showed the best effect on increasing tolerance to thermal and oxidative stress and reduce the accumulation of lipofuscin. In the assay of C. elegans, pb-3 reduced intracellular ROS accumulation. Application of pb-3 to the transgenic mutant TJ356 induced the translocation of the transcription factor DAF-16 from the cytosol to the nucleus, and modulated the expression of SOD-3 (downstream genes of daf-16), which regulates longevity in C. elegans. Moreover, pb-3 did not prolong the lifespan of daf-16, age-1, daf-2 and hsp16.2 mutants, suggesting that these genetic pathways are involved in mediating the antiaging effects of pb-3.Conclusion: The antioxidant and antiaging properties of pb-3 may involve in the DAF-16/FOXO transcription process. Pinocembrin amino acid derivatives might be a novel agent for antiaging therapy.Keywords: pinocembrin, antiaging, oxidative stress, amino acid derivatives, Caenorhabditis elegans, DAF-16/FOXO transcription factors

Abstract Background Oxidative stress is an important factor involved in the progress of heart failure. The current study was performed to investigate whether pinocembrin was able to ameliorate post-infarct heart failure (PIHF) and the underlying mechanisms. Methods Rats were carried out left anterior descending artery ligation to induce myocardial infarction and subsequently raised for 6 weeks to produce chronic heart failure. Then pinocembrin was administrated every other day for 2 weeks. The effects were evaluated by echocardiography, western blot, Masson’s staining, biochemical examinations, immunohistochemistry, and fluorescence. In vitro we also cultured H9c2 cardiomyocytes and cardiac myofibroblasts to further testify the mechanisms. Results We found that PIHF-induced deteriorations of cardiac functions were significantly ameliorated by administrating pinocembrin. In addition, the pinocembrin treatment also attenuated collagen deposition and augmented vascular endothelial growth factor receptor 2 in infarct border zone along with an attenuated apoptosis, which were related to an amelioration of oxidative stress evidenced by reduction of reactive oxygen species (ROS) in heart tissue and malondialdehyde (MDA) in serum, and increase of superoxide dismutase (SOD). This were accompanied by upregulation of nuclear factor erythroid 2-related factor 2 (Nrf2)/ heme oxygenase-1 (HO-1) pathway. In vitro experiments we found that specific Nrf2 inhibitor significantly reversed the effects resulted from pinocembrin including antioxidant, anti-apoptosis, anti-fibrosis and neovascularization, which further indicated the amelioration of PIHF by pinocembrin was in a Nrf2/HO-1 pathway-dependent manner. Conclusion Pinocembrin ameliorated cardiac functions and remodeling resulted from PIHF by ROS scavenging and Nrf2/HO-1 pathway activation which further attenuated collagen fibers deposition and apoptosis, and facilitated angiogenesis.