Objective To compare female C57BL/6J and BALB/c mice models of ulcerative colitis(UC)induced by dextran sulfate sodium salt(DSS),providing a reference for selecting female animal models in studies of innovative drugs for UC treatment.Methods A total of 48 female mice aged 6-8 weeks,including 24 C57BL/6J and 24 BALB/c mice,were divided into four groups:control group,UC-3d group(2.5%DSS drinking for 3 days),UC-7d group(2.5%DSS drinking for 7 days),and UC-withdrawal group(2.5%DSS drinking for 7 days followed by 3 days of pure water replacement),with 6 mice in each group.Body weight,fecal occult blood,and stool characteristics were recorded daily,and disease activity index(DAI)scores were calculated.At the end of DSS induction,mice were gavaged with fluorescein isothiocyanate(FITC)-labeled dextran solution before tissue collection.Four hours later,under deep anesthesia induced by inhalation of 4%isoflurane,blood samples were collected from the retro-orbital venous plexus,and euthanasia was performed by cervical dislocation.The colon was then separated immediately,and its length from the anus to the ileocecal junction was measured.Colon tissues were fixed,embedded,and sectioned,and they were evaluated using hematoxylin-eosin(HE)staining and alcian blue-periodic acid-Schiff(AB-PAS)staining to comprehensively assess the degree of colonic inflammation.The spleen was also collected and weighed,and the spleen coefficient was calculated.Serum FITC fluorescence intensity and D-lactic acid concentration were measured to evaluate intestinal permeability and barrier injury.The grooming duration,the number of buried marbles,the time spent in the center of the open field,and the total distance traveled were measured to evaluate anxiety/depression-like behaviors in mice.Results Fecal occult blood appeared in both C57BL/6J and BALB/c mice after 3 days of ad libitum access to DSS.Compared with the control group,there was no significant change in the weight loss rate of female BALB/c mice in the UC-7d and UC-withdrawal groups(P>0.05),but the DAI score increased(P<0.01).Conversely,both the weight loss rate and DAI score increased for female C57BL/6J mice in the UC-7d and UC-withdrawal groups(P<0.01).In female C57BL/6J mice,both the UC-7d and UC-withdrawal groups had higher spleen coefficients than the control group(P<0.05).Additionally,in these groups,colon length was significantly reduced(P<0.01),FITC fluorescence intensity in serum was markedly increased(P<0.01),obvious inflammatory reaction areas appeared in the colon,and the goblet cells and acidic mucus layer were damaged.In contrast,in female BALB/c mice,no significant differences were observed in colon length,serum FITC fluorescence intensity,or D-lactic acid concentration between the UC-7d and UC-withdrawal groups compared with the control group(P>0.05).Only a few inflammatory cells and damaged goblet cells were observed in the colonic mucosal layer,and no other obvious pathological changes were detected.In addition,female C57BL/6J mice in the UC-7d group showed a significantly reduced total distance in the open field and fewer buried marbles(P<0.05),indicating depression-like behavior,as well as prolonged grooming duration(P<0.05),indicating anxiety-like behavior.Conclusion Female BALB/c mice are not sensitive to 2.5%DSS and do not show typical colonic pathological changes of UC.In contrast,ad libitum access to 2.5%DSS for 7 days successfully induces typical UC pathological symptoms and colonic pathological changes in female C57BL/6J mice.These changes persist for 3 days after DSS withdrawal,and the mice also exhibit symptoms of anxiety/depression.The results suggest that this model can be used for research on the pathogenesis and therapeutic drugs of UC.

The annual output of Moutan Cortex is significant, but the epidermis of the root bark of peony (EPRP), as a by-product of Moutan Cortex, is typically discarded. To achieve full use of resources, this study aimed to develop an HPLC fingerprint analysis method using HPLC-DAD-ESI-MS/MS on EPRP sourced from different regions to establish EPRP quality control standards. For chromatography, a ShimPack Scepter C18 column (4.6 mm × 250 mm, 5 μm) was used. The mobile phase for HPLC consisted of acetonitrile (A) and a 0.1% aqueous formic acid solution (B). An HPLC fingerprint was established, featuring 30 characteristic peaks with a similarity of over 0.80. A total of 31 components were identified, with 22 chemical markers determined, including 1-galloylglucose, gallic acid, methyl gallate, oxypaeoniflora, paeonolide, apiopaeonoside, albiflorin, paeoniflorin, p-coumaric acid, ferulic acid, 1,2,3,6-tetragalloylglucose, ellagic acid, galloylpaeoniflorin, luteoloside, 1,2,3,4,6-O-penta-galloylglucose, diosmin, neodiosmin, resveratrol, mudanpioside C, benzoyloxypaeoniflorin, benzoylpaeoniflorin, and paeonol. These markers align with component structure theory, allowing for the analysis of the structural characteristics of EPRP from different regions. These findings provide a valuable reference for the future quality evaluation of EPRP, enhance the understanding of the components in EPRP from diverse sources, and lay a foundation for the development and greater utilization of EPRP.

This study investigated the therapeutic effects and mechanisms of Paeonia suffruticosa fruit pod extract (EMP) and its main component albiflorin (AF) on hyperuricemia-associated cognitive impairment (HUA-CI). A HUA-CI mouse model was established, with cognitive function evaluated via Morris water maze. Hippocampal pathology, inflammation, oxidative stress, and apoptosis were assessed using HE staining, ELISA, TUNEL, and Western blotting. Network pharmacology predicted EMP's targets, and molecular docking analyzed AF-MAP2K1 binding. In vitro experiments used UA-stimulated BV2 and HT22 cells to explore AF's effect on HIF-1 signaling. EMP significantly improved cognitive function and reduced pathological damage in the hippocampus of HUA-CI mice. It exerted protective effects by inhibiting inflammatory responses, alleviating oxidative stress, and preventing cell apoptosis. Network pharmacology analysis revealed that EMP acts through multiple targets and pathways, particularly via the strong binding affinity between AF and MAP2K1. Both in vivo and in vitro studies demonstrated that AF inhibited the HIF-1 signaling pathway, thereby reducing microglial activation and associated inflammation, mitigating uric acid-induced neuronal apoptosis, enhancing antioxidant defenses, and protecting neuronal function. Our research indicates that EMP exerts multi-target therapeutic effects on HUA-CI; AF plays a key role by targeting MAP2K1 and inhibiting HIF-1 signaling.

DCM is a serious complication of diabetes that eventually develops into heart failure and damages the health of human. Here, we study the therapeutic effect of MC-GF-CR on C57BL/6 mice with DCM. First, the aqueous extract powder of MC-GF-CR was characterized by HPLC for compositional analysis. Afterword, the network pharmacology method was adopted to gain the TCM-DCM network, PPI network, key targets, GO and KEGG enrichment analysis results. Meanwhile, we constructed the DCM mouse model by feeding high-sugar and high-fat fodder and injecting STZ to evaluate the therapeutic effect of MC-GF-CR on DCM and validate the potential targets. Thirteen chemicals were detected from the MC-GF-CR extracts by HPLC. 186 intersecting targets were screened between TCM and disease, including 31 core targets. The GO and KEGG analysis results suggested that MC-GF-CR can regulate DCM from various signaling pathways. The in vivo animal experiments exhibited that MC-GF-CR could improve the levels of body weight, blood glucose, blood lipids, serum insulin, cardiac function, and the expression of relative proteins in the model mice. Molecular docking results showed that IL-6-paeoniflorin and -quercetin, MAPK1-quercetin, and PRKCA-quercetin all exhibited a better binding ability. Our study reported that MC-GF-CR exhibited therapeutic effect on DCM through regulating the expression of IL-6, MAPK1, and PRKCA.

ETHNOPHARMACOLOGICAL RELEVANCE:The bark of Ailanthus altissima (Mill.) Swingle (BAA), a widely used Chinese medicinal herb in traditional remedies for bowel disorders, has yet to be explored in the context of ulcerative colitis (UC), and its therapeutic mechanisms remain unclear. AIM OF THE STUDY:This study integrated network pharmacology and experimental validation to investigate the effects and underlying mechanisms of BAA in treating UC. MATERIALS AND METHODS:First, UPLC-MS/MS analysis was employed to identify the chemical constituents of BAA. Network pharmacology was then applied to analyze the potential mechanisms of BAA based on these identified compounds. Lastly, a dextran sulfate sodium (DSS)-induced UC mouse model was utilized to assess BAA's therapeutic efficacy, with Western blotting performed to examine changes in protein expression within the key pathway influenced by BAA. RESULTS:UPLC-MS/MS and SwissADME analysis identified 223 active compounds in BAA. Network pharmacology suggested that the PI3K/AKT pathway may serve as a primary mechanism by which BAA exerts its anti-UC effects. In the DSS-induced UC mouse model, BAA significantly mitigated colonic injury, reduced DAI scores, and promoted weight recovery in mice. Additionally, BAA downregulated pro-inflammatory cytokines, including TNF-α, IL-1β, and IL-6, thereby suppressing inflammatory responses in the colon. Western blot analysis further demonstrated that BAA primarily inhibited the PI3K/AKT pathway in UC mouse colon tissue. CONCLUSION:This study highlights that BAA effectively reduces colonic inflammation and preserves intestinal mucosal integrity, likely through the inhibition of PI3K/AKT pathway activity, positioning it as a potential treatment for UC.

To evaluate the development and application of a kind of oil peony by chromatographic analysis of the main active components of the waste moutan pods (MP).The methods adopted in this paper, firstly, the quality evaluation method of MP is established by HPLC-DAD-ESI-MS/MS; The second is to compare the types and contents of the main active components of moutan barks (Chinese Pharmacopoeia, MB) with those of the medicinal parts of peony used in oil. HPLC fingerprint method for the extract of moutan pods (EMP) was established. Twenty-one compounds were identified in EMP and their levels determined in EMP and the extract of moutan barks (EMB) by HPLC. The results showed that the components of EMP and EMB were similar, though their contents differed. These findings suggest that EMP may have similar pharmacological effects to EMB. Our findings regarding the similarity and comparison of the chemical components in the EMB and EMP have important practical value for replacing EMB medicine and protecting peony plants from resource depletion caused by root digging.

ETHNOPHARMACOLOGICAL RELEVANCE:The Danshen-Shanzha Decoction (DSD) is a renowned herbal combination consisting of the root of Salvia miltiorrhiza Bunge (known as Danshen in Chinese) and the fruits of Crataegus pinnatifida Bunge (known as Shanzha in Chinese), which has exhibited remarkable clinical efficacy in the treatment of coronary heart disease (CHD) in traditional Chinese medicine, with its earliest recorded application dating to around 202 BCE during the Han Dynasty. Despite significant advancements in the fundamental research and clinical applications of DSD over the past few decades, the precise bioactive components as well as the underlying mechanisms responsible for its protective effect on CHD remain unelucidated. AIM OF THE STUDY:The present study was designed to elucidate the bioactive components and potential mechanism of DSD in the treatment of CHD using in silico technologies integrated with pharmacoinformatic methods and experimental validation. MATERIALS AND METHODS:The chemical components of DSD were analyzed and identified using UPLC-Q-TOF-MS. Pharmacoinformatic-based methods were employed to comprehensively investigate the principal active components and targets of DSD for treating CHD. GO and KEGG pathway analyses were utilized to elucidate the underlying mechanism responsible for DSD's efficacy against CHD. Molecular docking and molecular dynamics simulation were performed to assess the binding affinity between active components and putative targets. Furthermore, surface plasmon resonance (SPR) was carried out to verify the affinity and kinetic characteristics of major components to STAT3 protein. Subsequently, a series of in vitro experiments, including cell viability test, flow cytometric analysis, ELISA and western blotting, were conducted to validate the predicted results in an oxygen-glucose deprivation (OGD)-stimulated H9c2 model. RESULTS:A total of 96 compounds were characterized by UPLC-Q-TOF-MS, and 281 overlapping targets were identified through pharmacoinformatic-based methods. Among these, ten critical compounds were determined as the core active components of DSD. The core targets associated with the development of CHD included STAT3, SRC, TP53, JUN, and AKT1. Notably, Dihydrotanshinone I and (+)-Epicatechin exhibited strong binding affinity towards STAT3. The potential mechanisms by which DSD modulates the pathological progression of CHD were predicted to involve inflammation, oxidative stress, and apoptosis. Importantly, the cytoprotective effect of DSD against apoptosis was confirmed in OGD-stimulated H9c2 cells, as evidenced by the upregulation of Bcl-2 expression and downregulation of both Bax and cleaved caspase-3 expressions upon DSD treatment. Furthermore, DSD significantly enhanced the phosphorylated protein expressions of JAK2 and STAT3 compared to the OGD group, suggesting its potential role in modulating related signaling pathways. CONCLUSIONS:The current study successfully fills the gap in the understanding of the chemical profiles of DSD, predicting its active components, potential targets, and molecular mechanisms in the treatment of CHD. These findings not only provide a valuable strategy but also robust data support for future investigations into DSD, thereby facilitating the identification of novel therapeutic targets for traditional Chinese medicines in the battle against CHD.

Peptidyl arginine deiminase 4 (PAD4) is an enzyme playing essential role in many biological processes especially immune responses, and is a highly attractive therapeutic target. However, the number of PAD4 inhibitors is limited, and there is no inhibitor in clinical trial. This study aims to identify natural compounds holding high inhibitory effect on PAD4 activity from traditional herbal medicine. Of the thirty-six extracts of twelve medical plants, the dry root bark of Paeonia × suffruticosa Andrews (also known as Moutan Cortex) extracts showed highest inhibition effect on PAD4 activity. Further experiment was conducted with different batches of Moutan Cortex to confirm this result. In addition, we also evaluated the inhibitory activity of different parts of the Paeonia × suffruticosa Andrews, including flower ball, flower, leaf, pollen and seed meal. The results substantiated the hypothesis that Paeonia × suffruticosa Andrews contains compounds with high PAD4 inhibitory effect. Therefore, a practical bioactivity-guided fractionation coupling with a chemical profiling strategy was used to identify the fractions from Moutan Cortex extract with strong PAD4 inhibition activity, and the major constituents in these bioactive fractions were further characterized by LC-MS/MS. Among all the identified compounds, pentagalloylglucose (PGG) showed the most potent inhibition effect on PAD4 activity, with an IC50 value of 4.50 µM. The inhibition kinetic analysis, drug affinity response target stability (DARTS) and molecular docking experiments were performed to confirm the interaction between PGG and PAD4. The findings indicated that PGG functions as a mixed model PAD4 inhibitor. In summary, our results showed that PGG in Moutan Cortex is a highly potent PAD4 inhibitor, and PGG can be used as a promising lead compound for the development of effective PAD4 inhibitors.

Danshen-Shanzha Formula (DSF) is a well-known herbal combination comprising Radix Salvia Miltiorrhiza (known as Danshen in Chinese) and Fructus Crataegi (known as Shanzha in Chinese), It has been documented to exhibit considerable benefits for promoting blood circulation and removing blood stasis, and was used extensively in the treatment of atherosclerotic cardiac and cerebral vascular diseases over decades. Despite several breakthroughs achieved in the basic research and clinical applications of DSF over the past decades, there is a lack of comprehensive reviews summarizing its features and research, which hinders further exploration and exploitation of this promising formula. This review aims to provide a comprehensive interpretation of DSF in terms of its ethnopharmacological relevance, preparation methods, chemical constituents, pharmacokinetic properties and pharmacological effects. The related information on Danshen, Shanzha, and DSF was obtained from internationally recognized online scientific databases, including Web of Science, PubMed, Google Scholar, China National Knowledge Infrastructure, Baidu Scholar, ScienceDirect, ACS Publications, Online Library, Wan Fang Database as well as Flora of China. Data were also gathered from documentations, printed works and classics, such as the Chinese Pharmacopoeia, Chinese herbal classics, etc. Three essential avenues for future studies were put forward as follows: a) Develop and unify the standard preparation method of DSF as to achieve optimized pharmacological properties. b) Elucidate the functional mechanisms as well as the rationality and rule for the compatibility art of DSF by focusing on the clinic syndromes together with the subsequent development of preclinic study system in vitro and in vivo with consistent pathological features, pharmacokinetical behaviour and biomarkers. c) Perform more extensive clinical studies towards the advancement of mechanism-based on evidence-based medicine on the safety application of DSF. This review will provide substantial data support and broader perspective for further research on the renowned formula.

Background: Ailanthone, a small compound derived from the bark of Ailanthus altissima (Mill.) Swingle, has several anti-tumour properties. However, the activity and mechanism of ailanthone in colorectal cancer (CRC) remain to be investigated. This study aims to comprehensively investigate the mechanism of ailanthone in the treatment of CRC by employing a combination of network pharmacology, bioinformatics analysis, and molecular biological technique.Methods: The druggability of ailanthone was examined, and its targets were identified using relevant databases. The RNA sequencing data of individuals with CRC obtained from the Cancer Genome Atlas (TCGA) database were analyzed. Utilizing the R programming language, an in-depth investigation of differentially expressed genes was carried out, and the potential target of ailanthone for anti-CRC was found. Through the integration of protein-protein interaction (PPI) network analysis, GO and KEGG enrichment studies to search for the key pathway of the action of Ailanthone. Then, by employing molecular docking verification, flow cytometry, Transwell assays, and Immunofluorescence to corroborate these discoveries.Results: Data regarding pharmacokinetic parameters and 137 target genes for ailanthone were obtained. Leveraging The Cancer Genome Atlas database, information regarding 2,551 differentially expressed genes was extracted. Subsequent analyses, encompassing protein–protein interaction network analysis, survival analysis, functional enrichment analysis, and molecular docking verification, revealed the PI3K/AKT signaling pathway as pivotal mediators of ailanthone against CRC. Additionally, the in vitro experiments indicated that ailanthone substantially affects the cell cycle, induces apoptosis in CRC cells (HCT116 and SW620 cells), and impedes the migration and invasion capabilities of these cells. Immunofluorescence staining showed that ailanthone significantly inhibited the phosphorylation of AKT protein and suppressed the activation of the PI3K/AKT signaling pathway, thereby inhibiting the proliferation and metastasis of CRC cells.Conclusion: Therefore, our findings indicate that Ailanthone exerts anti-CRC effects primarily by inhibiting the activation of the PI3K/AKT pathway. Additionally, we propose that Ailanthone holds potential as a therapeutic agent for the treatment of human CRC.

Adding excipients is an important method to change the flavor and biological activity of food materials during processing. In this study, the contents of 11 bioactive compounds in the mulberry leaf tea with or without processing by addition of honey or salt, and their absorption and elimination characteristics in rats were determined. The biological activities of processed products were studied by in vitro models, and the effects of different processing methods on the compounds and biological activities of mulberry leaf tea extracts were analyzed by multiple factor analysis. We found that different processing methods can change the contents of some compounds in mulberry leaf tea extracts, and then affect the biological activity of extracts. The processing method of adding honey and salt can respectively enhance the antioxidant capacity and anti-apoptotic effect of mulberry leaf, while the processing method without auxiliary materials was more conducive to the repair of blood vessels.

Mulberry (Morus alba L.) has a special significance in the history of agriculture and economic plant cultivation. Mulberry has strong environmental adaptability, a wide planting range, and abundant output. It is not only an important resource for silkworm breeding but also a raw ingredient for various foods and has great potential for the development of biological resources. The bioactivities of mulberry in different planting areas are not the same, which is an obstacle to the development of mulberry. This study collected information on the planting conditions of mulberry branches in 12 planting areas, such as altitude, temperature difference, and precipitation. A comparison of the levels of 12 constituents of mulberry branches from mulberry grown in different planting areas was then made. An in vitro model was used to study the bioactivities of mulberry branches in the 12 planting areas, and mathematical analysis was used to explain the possible reasons for the differences in the composition and bioactivities of mulberry branches in different planting areas. After studying mulberry samples from 12 planting areas in China, it was found that a small temperature difference could affect the antiapoptotic effect of mulberry branch on microvascular endothelial cells by changing the levels and proportions of rutin, hyperoside, and morusin. Adequate irrigation can promote the antioxidation of the mulberry branch on microvascular endothelial cells by changing the levels and proportions of scopoletin and quercitrin. The results of the analysis of planting conditions and the levels of active constituents and their correlation with bioactivities support the improvement of mulberry planting conditions and have great significance in the rational development of mulberry resources. This is the first time that a mathematical analysis method was used to analyze the effects of planting conditions on mulberry biological activity.

Ethnopharmacological relevance: Ilex rotunda Thunb. (IR) is widely used for gastrointestinal diseases by Yao physician, and it has a better clinical curative effect on ulcerative colitis (UC). However, the main active components and mechanism of IR in the treatment of UC remain to be clarified.Aim of the study: To investigate the main active components and mechanism of IR in the treatment of UC.Materials and methods: Ten biological active components of IR were quantified by UPLC-MS/MS. In vitro, Caco2 cell monolayers were stimulated by lipopolysaccharide, and were treated with 10 biologically active components individually to investigate the protective role of the components of IR in mucosal barrier damage. In vivo, a mouse model of UC was induced by dextran sulfate sodium and administered with the candidate active components of IR. On day 8, the serum and colon tissue were collected for histological and molecular analysis to investigate the main active components and mechanism of IR.Results: Ziyuglycoside I, ziyuglycoside II, syringin, and pedunculoside in IR reduced phenol red transmission of the monolayer, and inhibited the protein expression of oncostatin M and oncostatin M receptor in Caco2 cells. Notably, ziyuglycoside II and syringin decreased the transepithelial electrical resistance of the monolayer, and promoted the protein expression of Occludin, Claudin-1 and zonula occludens-1 (ZO-1) in Caco2 cells. In vivo, ziyuglycoside II and syringin improved the symptoms of UC mice, including body weight, disease activity score, shortening of colon length, damaging of acidic mucus layer, histopathological changes, and protein expression of Occludin, Claudin-1, and ZO-1. Pedunculoside reduced the neutrophils and inflammatory response in the UC mice. Moreover, when the combination of ziyuglycoside II, syringin and pedunculoside was used for the treat-ment of UC, syringin and pedunculoside enhanced the therapeutic effect of ziyuglycoside II. Finally, RNA sequencing and RT-qPCR analysis revealed that ziyuglycoside II + syringin + pedunculoside and IR coregulated up to 42.7% of genes, and mainly reduced the overexpression of C-X-C motif ligand 1(CXCL1), oncostatin M receptor (OSMR), interleukin 1 receptor type I (IL1R1), tumor necrosis factor receptor superfamily member 9 (TNFRSF9), C-X-C motif chemokine 13 (CXCL13), oncostatin M (OSM), and interleukin 6 (IL-6) in the cytokine-cytokine interaction pathways.Conclusions: The combination of ziyuglycoside II, syringin, and pedunculoside protects against UC by modulating the intestinal mucosal barrier and inhibiting the cytokine-cytokine interaction pathways, and the effect is relatively equivalent to that of the water extract of Ilex rotunda Thunb.

目的 探讨四季抗病毒合剂(Siji Kangbingdu Mixture,SJM)及其主成分(principal component,PC:连翘酯苷 A、连翘酯苷B、连翘苷、甘草苷、甘草酸、木犀草苷、绿原酸、1,3-二咖啡酰奎宁酸、3,5-二咖啡酰奎宁酸)抗肠道病毒 71 型(enterovirus 71,EV71)增殖和感染的作用及其作用机制.方法 MTS法检测 SJM、PC 对非洲绿猴肾细胞(Vero)的毒性;基于 EV71 感染细胞模型检测 SJM、PC抗损伤的效果;基于 EV71 感染乳鼠模型观察 SJM、PC对其存活率、存活时间、临床症状评分的改善情况;检测乳鼠模型肌肉与血清中白细胞介素-6(interleukin 6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、单核细胞趋化蛋白-1(monocyte chemoattractant protein-1,MCP-1)水平.结果 (1)EV71对 Vero 细胞的半数组织培养感染剂量(TCID50)为10-5.68·mL-1;(2)SJM、PC对 Vero细胞的半数抑制质量浓度分别为11.83 mg·mL-1(按生药量计)、20.67μg·mL-1(按连翘酯苷A计);(3)单次和连续给予 SJM与PC对 ICR乳鼠无明显致毒作用;(4)SJM与 PC 可显著抑制 EV71 的增殖,以提高被感染细胞的存活率;(5)SJM与PC可显著改善被感染乳鼠的存活率、临床症状等指标;(6)SJM与PC均可显著调节肌肉与血清中IL-6、TNF-α、MCP-1 的水平.结论 SJM与PC可显著抑制 EV71 的增殖,调节免疫炎症因子,PC可在一定程度上替代 SJM,发挥治疗手足口病的重要作用.

Paeonia suffruticosa is widely cultivated globally due to its medicinal and ornamental value. Peony pollen (PP) is commonly used in Chinese folk medicine to make tea to treat benign prostatic hyperplasia (BPH), but its molecular mechanism against BPH is yet to be comprehended. The objective of this research was to experimentally verify the effect of PP in the treatment of BPH and to preliminarily reveal its mechanism of action on BPH using network pharmacology methods. The results revealed that PP could decrease prostate volume and prostate index, serum testosterone (T), dihydrotestosterone (DHT), and estradiol (E2) levels. Moreover, it could improve prostate tissue structure in BPH model animals as well. Additionally, database searches and disease target matching revealed 81 compounds in PP. Of these, 3, 7, 8, 2′-tetrahydroxyflavone, Chrysin, Wogonin, Limocitrin, and Sexangularetin were the top five compounds associated with the therapeutic effects of BPH. Furthermore, 177 therapeutic targets for BPH were retrieved from databases of Swiss Target, DisGeNET, Drugbank, Genecards, OMIM, TTD, and Uniprot. In contrast, core targets AKT1, EGFR, IL6, TNF, and VEGFA were obtained by PPI network diagram. Molecular docking also showed that the main efficacy components and potential core targets in PP had good binding capacity. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomics (KEGG) analysis established that the effect of PP in BPH therapy was mainly through regulating the expression levels of protein kinase B on phosphatidylinositol 3-kinase and phosphatidylinositol 3-kinase-protein kinase B pathways. Additionally, Western blot experiments also exhibited a significant elevation in the activated PI3K and AKT proteins in the model (Mod) group relative to the control (Con) group, and the expression of these activated proteins was significantly reduced after PP administration. In summary, this research provides a scientific basis for employing PP to treat BPH, preliminarily reveals its mechanism of action and potential targets, and lays the foundation for further research and development.

Background The dried bark of Ailanthus altissima (Mill.) Swingle is widely used in traditional Chinese medicine for the treatment of ulcerative colitis. The objective of this study was to explore the therapeutic basis of the dried bark of Ailanthus altissima (Mill.) Swingle for the treatment of ulcerative colitis based on Virtual Screening–Molecular Docking–Activity Evaluation technology. Methods By searching the Traditional Chinese Medicine Systems Pharmacology TCMSP Database and Analysis Platform, 89 compounds were obtained from the chemical components of the dried bark of Ailanthus altissima (Mill.) Swingle. Then, after preliminarily screening the compounds based on Lipinski’s rule of five and other relevant conditions, the AutoDock Vina molecular docking software was used to evaluate the affinity of the compounds to ulcerative colitis-related target proteins and their binding modes through use of the scoring function to identify the best candidate compounds. Further verification of the compound’s properties was achieved through in vitro experiments. Results Twenty-two compounds obtained from the secondary screening were molecularly docked with ulcerative colitis-related target proteins (IL-1R, TLR, EGFR, TGFR, and Wnt) using AutoDock Vina. The free energies of the highest scoring compounds binding to the active cavity of human IL-1R, TLR, EGFR, TGFR, and Wnt proteins were − 8.7, − 8.0, − 9.2, − 7.7, and − 8.5 kcal/mol, respectively. The potential compounds, dehydrocrebanine, ailanthone, and kaempferol, were obtained through scoring function and docking mode analysis. Furthermore, the potential compound ailanthone (1, 3, and 10 µM) was found to have no significant effect on cell proliferation, though at 10 µM it reduced the level of pro-inflammatory factors caused by lipopolysaccharide. Conclusion Among the active components of the dried bark of Ailanthus altissima (Mill.) Swingle, ailanthone plays a major role in its anti-inflammatory properties. The present study shows that ailanthone has advantages in cell proliferation and in inhibiting of inflammation, but further animal research is needed to confirm its pharmaceutical potential.

Background: Hyperuricemia is an important pathological basis of gout and a distinct hazard factor for metabolic syndromes and cardiovascular and chronic renal disease, but lacks safe and effective treatments currently. Paeonia x suffruticosa Andrews leaf effectively reduced serum uric acid in gout patients; however, the material foundation and the mechanism remain unclear. Purpose: To determine the primary active components and mechanism of P. suffruticosa leaf in hyperuricemic mice. Methods: The chemical constituents of P. suffruticosa leaf was identified using high-performance liquid chro-matographic analysis. The anti-hyperuricemic activity of P. suffruticosa leaf extract (12.5, 25, 50, 100, and 200 mg/kg) and its components was evaluated in hyperuricemic mice induced by a high purine diet for 14 days. Then, the urate-lowering effects of apigenin 7-O-glucoside (0.09, 0.18, and 0.36 mg/kg) were assessed in another hyperuricemic mice model built by administrating potassium oxonate and adenine for 4 weeks. The inhibitory effect of apigenin 7-O-glucoside on uric acid production was elucidated by investigating xanthine oxidase activity in vitro and in serum and the liver and through molecular docking. Immunofluorescence and western blot an-alyses of the expression of renal urate transporter 1 (URAT1), glucose transporter 9 (GLUT9), organic anion transporters 1 (OAT1), and ATP-binding cassette G member 2 (ABCG2) proteins elucidated how apigenin 7-O-glucoside promoted uric acid excretion. Results: Six compounds were identified in P. suffruticosa leaf: gallic acid, methyl gallate, oxypaeoniflorin, paeoniflorin, galloylpaeoniflorin, and apigenin 7-O-glucoside. P. suffruticosa leaf extract significantly attenuated increased serum uric acid, creatinine, and xanthine oxidase activity in hyperuricemic mice. Apigenin 7-O-glucoside from P. suffruticosa leaf reduced uric acid, creatinine, and malondialdehyde serum levels, increased superoxide dismutase activity, and partially restored the spleen coefficient in hyperuricemic mice. Apigenin 7-O-glucoside inhibited xanthine oxidase activity in vitro and decreased serum and liver xanthine oxidase activity and liver xanthine oxidase protein expression in hyperuricemic mice. Molecular docking revealed that apigenin 7-O-glucoside bound to xanthine oxidase. Apigenin 7-O-glucoside facilitated uric acid excretion by modulating the renal urate transporters URAT1, GLUT9, OAT1, and ABCG2. Apigenin 7-O-glucoside protected against renal damage and oxidative stress caused by hyperuricemia by reducing serum creatinine, blood urea nitrogen, malondialdehyde, and renal reactive oxygen species levels; increasing serum and renal superoxide dismutase activity; restoring the renal coefficient; and reducing renal pathological injury. Conclusion: Apigenin 7-O-glucoside is the main urate-lowering active component of P. suffruticosa leaf extract in the hyperuricemic mice. It suppressed liver xanthine oxidase activity to decrease uric acid synthesis and modulated renal urate transporters to stimulate uric acid excretion, alleviating kidney damage caused by hyperuricemia.

Objective:To establish a method for the simultaneous determination of O-nornuciferine, quercetin-3-O-sycambinoside, hyperoside, isoquercitrin, quercetin-3-O-glucuronide, and nuciferine in water extract of lotus leaf. Methods:The column was Lamdo Stamsil C 18 （250 mm×4.6 mm, 5 μm）, the mobile phase was 0.1% phosphoric acid aqueous solution-acetonitrile in gradient mode at a flow rate of 0.6 mL·min -1 . The detective wavelength was set at 270 nm. The column temperature was 25 ℃ and the injection volume was 5 μL. The relative correction factors of the other five components to O-nornuciferine were calculated and their contents were determined. Results:The linear relationships of O-nornuciferine, quercetin-3-O-sambulide, hyperoside, isoquercitrin, quercetin-3-O-glucuronide, and nuciferine were good（r≥0.999 5）. The RSDs of precision, repeatability and stability tests were less than 3.0%. The average recovery was 94.6%-108.0% with RSDs within 0.95%-2.1%. The content of the six components in the water extract of lotus leaf was determined by QAMS was 0.23, 3.43, 1.69, 10.59, 41.23, 0.69 mg·g -1 . Conclusion:The established method for the determination of 6 components in the water extract of lotus leaf is reliable and can be used for its quality control and pharmacodynamics research.

Peony pollen contains multiple nutrients and components and has been used as a traditional Chinese medicine with a long history, but the effect of the treatment of primary dysmenorrhea remains to be clarified. The aim of this study is to investigate the therapeutic effect of peony pollen on primary dysmenorrhea mice and the potential mechanism. A uterus contraction model in vitro and primary dysmenorrhea mice were used to evaluate the treatment effect of peony pollen on primary dysmenorrhea. The primary dysmenorrhea mice were treated with 62.5 mg/kg, 125 mg/kg, or 250 mg/kg of peony pollen, and the writhing response, latency period, histopathological changes in the uterus, prostaglandin E2 (PGE2) and prostaglandin F2α (PGF2α) levels, and infiltration of neutrophils and macrophages were investigated. Protein expression of interleukin 1 β (IL-1β), interleukin 6 (IL-6), NOD-like receptor thermal protein domain associated protein 3 (NLRP3), cyclooxygenase-2 (COX-2), microsomal prostaglandin-E synthase 1 (mPGEs-1), BCL2-Associated X (Bax), B-cell lymphoma-2 (BCL-2), caspase-3, and cleaved caspase-3 were detected by Western blot, and the oxidative stress related marker malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and reactive oxygen species (ROS) were evaluated. Peony pollen could attenuate spontaneous or oxytocin-induced uterus contractions in vitro. Moreover, peony pollen decreased the writhing times, prolonged the writhing latency, and reduced the pathological damage of uterine tissues. Furthermore, the inflammatory cell infiltration and the protein expression of IL-1β, IL-6, and NLRP3 were decreased. The COX-2/PGE2 pathway was inhibited; oxidative stress and apoptosis in the uterus also improved in the uterus of primary dysmenorrhea mice. Peony pollen exerts a positive effect on primary dysmenorrhea by inhibiting the inflammatory response and modulating oxidative stress and apoptosis by regulating the COX-2/PGE2 pathway.

Coronavirus disease 2019 (COVID‐19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2). In China, the Acacia catechu (AC)‐Scutellariae Radix (SR) formula has been widely used for pulmonary infection in clinical practice for several centuries. However, the potential role and mechanisms of this formula against COVID‐19 remains unclear. The present study was designed to dissect the active ingredients, molecular targets, and the therapeutic mechanisms of AC‐SR formula in the treatment of COVID‐19 based on a systems pharmacology strategy integrated by ADME screening, target prediction, network analysis, GO and KEGG enrichment analysis, molecular docking, and molecular dynamic (MD) simulations. Finally, Quercetin, Fisetin(1‐), kaempferol, Wogonin, Beta‐sitosterol, Baicalein, Skullcapflavone II, Stigmasterol were primarily screened to be the potentially effective active ingredients against COVID‐19. The hub‐proteins were TP53, JUN, ESR1, MAPK1, Akt1, HSP90AA1, TNF, IL‐6, SRC, and RELA. The potential mechanisms of AC‐SR formula in the treatment of COVID‐19 were the TNF signaling pathway, PI3K‐Akt signaling pathway and IL‐17 signaling pathway, etc. Furthermore, virtual docking revealed that baicalein, (+)‐catechin and fisetin(1‐) exhibited high affinity to SARS‐CoV‐2 3CLpro, which has validated by the FRET‐based enzymatic inhibitory assays with the IC50 of 11.3, 23.8, and 44.1 μM, respectively. And also, a concentration‐dependent inhibition of baicalein, quercetin and (+)‐catechin against SARS‐CoV‐2 ACE2 was observed with the IC50 of 138.2, 141.3, and 348.4 μM, respectively. These findings suggested AC‐SR formula exerted therapeutic effects involving “multi‐compounds and multi‐targets.” It might be working through directly inhibiting the virus, improving immune function, and reducing the inflammatory in response to anti‐COVID‐19. Ultimately, this study would provide new perspective for discovering potential drugs and mechanisms against COVID‐19.