Huangqin decoction alleviates lipid metabolism disorders and insulin resistance in nonalcoholic fatty liver disease by triggering Sirt1/NF-κB pathway

BACKGROUND Nonalcoholic fatty liver disease(NAFLD)is a clinicopathological entity characterized by intrahepatic ectopic steatosis.As a consequence of increased consumption of high-calorie diet and adoption of a sedentary lifestyle,the incidence of NAFLD has surpassed that of viral hepatitis,making it the most common cause of chronic liver disease globally.Huangqin decoction(HQD),a Chinese medicinal formulation that has been used clinically for thousands of years,has beneficial outcomes in patients with liver diseases,including NAFLD.However,the role and mechanism of action of HQD in lipid metabolism disorders and insulin resistance in NAFLD remain poorly understood.AIM To evaluate the ameliorative effects of HQD in NAFLD,with a focus on lipid metabolism and insulin resistance,and to elucidate the underlying mechanism of action.METHODS High-fat diet-induced NAFLD rats and palmitic acid(PA)-stimulated HepG2 cells were used to investigate the effects of HQD and identify its potential mechanism of action.Phytochemicals in HQD were analyzed by highperformance liquid chromatography(HPLC)to identify the key components.RESULTS Ten primary chemical components of HQD were identified by HPLC analysis.In vivo,HQD effectively prevented rats from gaining body and liver weight,improved the liver index,ameliorated hepatic histological aberrations,decreased transaminase and lipid profile disorders,and reduced the levels of pro-inflammatory factors and insulin resistance.In vitro studies revealed that HQD effectively alleviated PA-induced lipid accumulation,inflammation,and insulin resistance in HepG2 cells.In-depth investigation revealed that HQD triggers Sirt1/NF-κB pathwaymodulated lipogenesis and inflammation,contributing to its beneficial actions,which was further corroborated by the addition of the Sirt1 antagonist EX-527 that compromised the favorable effects of HQD.CONCLUSION In summary,our study confirmed that HQD mitigates lipid metabolism disorders and insulin resistance in NAFLD by triggering the Sirt1/NF-κB pathway.

Network pharmacology and molecular docking-based analyses to predict the potential mechanism of Huangqin decoction in treating colorectal cancer

BACKGROUND To analyze the potential action mechanism of Huangqin decoction(HQD)in colorectal cancer(CRC)treatment on the basis of network pharmacology and molecular docking.AIM To investigate the molecular mechanisms of HQD for CRC treatment by using network pharmacology and molecular docking.METHODS All HQD active ingredients were searched using the Systematic Pharmacology and Traditional Chinese Medicine Systems Pharmacology databases and the Bioinformatics Analysis Tool for Molecular Mechanisms in traditional Chinese medicine.Then,the targets of the active ingredients were screened.The abbreviations of protein targets were obtained from the UniProt database.A“drug–compound–target”network was constructed to screen for some main active ingredients.Some targets related to the therapeutic effect of CRC were obtained from the GeneCards,DisGeNET,Therapeutic Target Database,and Online Mendelian Inheritance in Man databases.The intersection of targets of Chinese herbs and CRC was taken.A Venn diagram was drawn to construct the intersection target interactions network by referring to the STRING database.Topological analysis of the protein interaction network was performed using Cytoscape 3.7.2 software to screen the core HQD targets for CRC.The core targets were imported into the DAVID 6.8 analysis website for gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses and visualization.Finally,molecular docking was performed using AutoDockTool and PyMOL for validation.RESULTS In total,280 potential drug-active ingredients were present in HQD,including 1474 targets of the drug-active ingredients.The main active ingredients identified were betulin,tetrahydropalmatine,and quercetin.In total,10249 CRC-related targets and 1014 drug-disease intersecting targets were identified,including 28 core targets of action such as Jun proto-oncogene,AP-1 transcription factor subunit,signal transducer and activator of transcription 3,tumor protein p53,vascular endothelial growth factor,and AKT serine/threonine kinase 1.The gene ontology enrichment functional analysis yielded 503 enrichment results,including 406 biological processes that were mainly related to the positive regulation of both gene expression and transcription and cellular response to hypoxia,etc.In total,38 cellular components were primarily related to polymer complexes,transcription factor complexes,and platelet alpha granule lumen.Then,59 molecular functions were closely related to the binding of enzymes,homologous proteins,and transcription factors.The Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis yielded 139 enrichment results,involving epidermal growth factor receptor tyrosine kinase inhibitor resistance and HIF-1 and mitogen-activated protein kinase signaling pathways.CONCLUSION HQD can play a role in CRC treatment through the“multi-component-target–pathway”.The active ingredients betulin,tetrahydropalmatine,and quercetin may act on targets such as Jun proto-oncogene,AP-1 transcription factor subunit,signal transducer and activator of transcription 3,tumor protein p53,vascular endothelial growth factor,and AKT serine/threonine kinase 1,which in turn regulate HIF-1 and mitogen-activated protein kinase signaling pathways in CRC treatment.The molecular docking junction clarified that all four key target proteins could bind strongly to the main HQD active ingredients.This indicates that HQD could slow down CRC progression by modulating multiple targets and signaling pathways.

Network pharmacology approach on the mechanism of Sanwu Huangqin Decoction in treating Behcet's disease

Objective:To analyze the main compounds of Sanwu Huangqin decoction based on the method of network pharmacology and to predict its possible target and mechanism of action in the treatment of Behcet's disease(BD).Methods:TCMSP,TCMID and literatures were used to search the main compounds and their corresponding targets of Sanwu Huangqin Decoction;the disease targets of BD were searched by GeneCards database;the intersection targets of Sanwu Huangqin Decoction and BD were visualized by String database and Cytoscape software;the intersection targets were enriched by GO and KEGG by DAVID database;BD mouse model was established,RT-PCR was used to verify the regulatory effect of Sanwu Huangqin Decoction on the core target genes.Results:A total of 81 main active compounds and 50 drug-disease-target were retrieved from the databases.There were 531 biological function information and 96 signal pathways were enriched by GO and KEGG.Among them,the top 5 signal pathways were Fluid shear stress and atherosclerosis、AGE-RAGE signaling pathway in diabetic complications、IL-17 signaling pathway、TNF signaling pathway、Malaria、Chagas disease(American trypanosomiasis).RT-PCR results showed that the expression of IL-4,IL-1β,IL-6,INF-γmRNA in the spleen of BD mice was significantly up-regulated,and the expression of Caspase 3 and Caspase 8 mRNA was significantly down regulated when compared with the control group(P<0.01).Compared with the model group,the expression of IL-4,IL-1β,IL-6,INF-γmRNA in the Sanwu Huangqin decoction group was significantly down regulated,and the expression of caspase 3 and caspase 8 mRNA was significantly up regulated(P<0.01).Conclusion:Sanwu Huangqin decoction can treat BD through multi-target and multi-channel therapy,and our study and our research provides bioinformatics basis for further revealing its mechanism and treatment.

Effect of Sanwu Huangqin Decoction on IFN-γ and IL-17A levels in mice with Behcet's disease

Objective:To investigate the effect of Sanwu Huangqin Decoction on IFN-γand IL-17A levels in mice with Behcet's disease;Methods:Clinical samples of patients with Behcet's disease were collected and mononuclear cells were isolated.High throughput RNA-seq was used to identify the different genes between Behcet's disease and healthy subjects.Mouse model was constructed by shrimp myosin sensitization and treated with Sanwu Huangqin decoction.The levels of IFN-γand IL-17A in peripheral blood and mononuclear cells were detected by ELISA and flow cytometry.The expression level of TBX21 gene in spleen was detected by RT-PCR;Results:Compared with healthy subjects,31 differential genes were identified in BD patients,of which 19 genes were up-regulated and 12 genes were down regulated.PPI analysis showed that the top 5 genes were TBX21,STAT3,IL15,PRF1 and IFNG.The results of animal experiments showed that the levels of IFN-γand IL-17A in the peripheral blood and PBMC of the model group were significantly higher compared with the control group(P<0.01).Sanwu Huangqin decoction could significantly reduce the levels of IFN-γand IL-17A in the peripheral blood and PBMC of mice(P<0.05 or P<0.01).RT-PCR results showed that the expression of TBX21 mRNA in spleen tissue of model group was significantly higher than that of the control group(P<0.01).Sanwu Huangqin decoction could significantly down-regulate the expression of TBX21 mRNA in spleen(P<0.05 or P<0.01);Conclusion:Sanwu Huangqin decoction reduce the levels of IFN-γand IL-17A by inhibiting the expression of TBX21 mRNA,so as to achieve the effect of treating BD.

A Network Pharmacology Approach to Investigate the Mechanisms of Huangqin Decoction in the Treatment of Irinotecan-Induced Gastrointestinal Toxicity

Objective:To find out the potential mechanisms of Huangqin decoction in the treatment of irinotecan-induced gastrointestinal toxicity.Methods:A network pharmacology approach was used to analyze the active compounds,drug targets and interacting pathways of Huangqin decoction in treating irinotecan-induced gastrointestinal toxicity.The compounds and predicted targets of Huangqin decoction were screened from TCMSP,and the disease targets were obtained from GeneCards.The therapeutic mechanisms of action of the Huangqin decoction were analyzed by gene ontology(GO)enrichment,Kyoto encyclopedia of genes and genomes pathway(KEGG)enrichment analyses.Results:The results show that 161compounds and 143 targets were obtained in this work.These targets were further mapped to 216 GO biological process terms and 30 remarkably pathways.Active compounds,targets,and pathways were used to construct a compound-target network.These results indicated that Huangqin decoction may treat the irinotecan-induced gastrointestinal toxicity mainly from intervening in the mucosal inflammation,cell apoptosis process,and cell proliferation.Conclusion:This study confirmed that the active components of Huangqin decoction play an important role in the treatment of irinotecan-related gastrointestinal toxicity through multi-target and multi-pathway,which provides a new way for the pathogenesis of irinotecan-related gastrointestinal toxicity.It facilitates the modernization of herbal medicine for complex diseases in the future.

Huangqin Decoction Delays Progress of Colitis-Associated Carcinogenesis by Regulating Nrf2/HO-1 Antioxidant Signal Pathwayin Mice

Objective:To investigate the effect of Huangqin Decoction(HQD)on nuclear factor erythroid 2 related-factor 2(Nrf2)/heme oxygenase(HO-1)signaling pathway by inducing the colitis-associated carcinogenesis(CAC)model mice with azoxymethane(AOM)/dextran sodium sulfate(DSS).Methods:The chemical components of HQD were analyzed by liquid chromatography-quadrupole-time-of-flight mass spectrometry(LC-Q-TOF-MS/MS)to determine the molecular constituents of HQD.Totally 48 C57BL/6J mice were randomly divided into 6 groups by a random number table,including control,model(AOM/DSS),mesalazine(MS),low-,medium-,and high-dose HQD(HQD-L,HQD-M,and HQD-H)groups,8 mice in each group.Except for the control group,the mice in the other groups were intraperitoneally injected with AOM(10 mg/kg)and administrated with 2.5%DSS orally for 1 week every two weeks(totally 3 rounds of DSS)to construct a colitis-associated carcinogenesis mouse model.The mice in the HQD-L,HQD-M and HQD-H groups were given HQD by gavage at doses of 2.925,5.85,and 11.7 g/kg,respectively;the mice in the MS group was given a suspension of MS at a dose of 0.043 g/kg(totally 11 weeks).The serum levels of malondialdehyde(MDA)and superoxide dismutase(SOD)were measured by enzyme-linked immunosorbent assay.The mRNA and protein expression levels of Nrf2,HO-1,and inhibitory KELCH like ECH-related protein 1(Keap1)in colon tissue were detected by quantitative real-time PCR,immunohistochemistry,and Westem blot,respectively.Results:LC-Q-TOF-MS/MS analysis revealed that the chemical constituents of HQD include baicalin,paeoniflorin,and glycyrrhizic acid.Compared to the control group,significantly higher MDA levels and lower SOD levels were observed in the model group(P<0.05),whereas the expressions of Nrf2 and HO-1 were significantly decreased,and the expression of Keap1 increased(P<0.01).Compared with the model group,serum MDA level was decreased and SOD level was increased in the HQD-M,HQD-H and MS groups(P<0.05).Higher expressions of Nrf2 and HO-1 were observed in the HQD groups.Conclusion:HQD may regulate the expression of Nrf2 and HO-1 in colon tissue,reduce the expression of MDA and increase the expression of SOD in serum,thus delaying the progress of CAC in AOM/DSS mice.

Huangqin Decoction Attenuates DSS-Induced Mucosal Damage and Promotes Epithelial Repair via Inhibiting TNF-α-Induced NF-κB Activation

Objective:To investigate the protective effect of Chinese herbal formula Huangqin Decoction(HQD)on ulcerative colitis mouse model induced by dextran sulphate sodium(DSS)and human intestinal epithelial cell injury induced by tumour necrosis factor-α(TNF-α).Methods:In vivo,30 male C57BL/6 mice were divided into 5 groups using a random number table(n=6 per group),including control,DSS,5-aminosalicylic acid(5-ASA),HQD low-(HQD-L)and high-dose(HQD-H)groups.The colitis mouse model was established by 3%(w/v)DSS water for 5 days.Meanwhile,mice in the HQD-L,HQD-H and 5-ASA groups were administrated with 100,200 mg/kg HQD or 100 mg/kg 5-ASA,respectively,once daily by gavage.After 9 days of administration,the body weight,disease activity index(DAI)score and colon length of mice were measured,the pathological changes of colons were analyzed by hematoxylin-eosin staining(HE)staining,and the levels of serum interleukin(IL)-6,IL-1βand TNF-αwere measured by enzyme linked immunosorbent assay.In vitro,the human colon epithelial normal cells(FHC cells)were exposed to HQD(0.6 mg/mL)for 12 h and then treated with TNF-α(10 ng/mL)for 24 h.The tight junction(TJ)protein expression levels of Claudin-4 and Occludin,and the protein phosphorylation levels of p65 and inhibitor of nuclear factor kappaB(NF-κB)-α(IκBα)were measured by Western blot.Results:In vivo,compared with the DSS group,HQD-H treatment attenuated the weight loss and reduced DAI score of mice on the 8th day(P<0.05).Moreover,HQD-H treatment ameliorated the colon shortening in the DSS-induced colitis mice(P<0.05).HE staining showed HQD attenuated the pathological changes of colitis mice,and the histological scores of HQD-H and 5-ASA groups were significantly decreased compared with the DSS group(P<0.05).Meanwhile,HQD-H and 5-ASA significantly decreased the serum IL-1β,IL-6 and TNF-αlevels of mice(P<0.05).In vitro experiments showed that HQD up-regulated Occludin and Claudin-4 protein expressions and inhibited p-p65 and p-IκBαlevels in FHC cells compared with the TNF-αgroup(P<0.05).Conclusion:HQD significantly relieved the symptoms in DSS-induced colitis mice by inhibiting pro-inflammatory cytokines expression and maintained the homeostasis of TJ protein in FHC cells by suppressing TNF-α-induced NF-κB activation.