山东医药2025，Vol.65Issue(3)：1-6,6.DOI:10.3969/j.issn.1002-266X.2025.03.001
柴胡疏肝散治疗肝纤维化核心靶基因筛选及对肝星状细胞活化的影响
Screening of core target genes of Chaihu Shugan powder in treatment of hepatic fibrosis and its effect on activation of hepatic stellate cells
摘要
Abstract
Objective To screen core target genes of Chaihu Shugan powder(CSP)in treatment of hepatic fibrosis via network pharmacology and to analyze its effect and mechanism on the activation of hepatic stellate cells.Methods The target genes of the active ingredients in CSP were intersected with the target genes of liver fibrosis to obtain the potential anti-liver fibrosis target genes of CSP.Gene Ontology(GO),KEGG,and Reactome pathway analyses were performed.The protein-protein interaction network of the potential anti-liver fibrosis target genes of CSP was constructed using STRING platform,and the core target genes were screened by degree value.Human hepatic stellate cells LX-2 were divid-ed into the transforming growth factor-β1(TGF-β1)group,low-dose group,medium-dose group,high-dose group(treated with 2 ng/mL TGF-β1 and 0,12.5,25,and 50 mg/mL CSP for 24 h),and the Control group(untreated).Real-time PCR and Western blotting were employed to detect the mRNA and protein levels of fibrotic markers,including COL1A1,vimentin,and α-SMA.Immunofluorescence was used to detect the expression of Fibronectin(represented by green fluores-cence intensity).LX-2 cells were divided into the pcDNA3.1 group(transfected with pcDNA3.1 plasmid),the signal transducer and activator of transcription 3(STAT3)-Flag group(transfected with pcDNA3.1-STAT3-Flag plasmid),and the Control group(untreated).After transfection,the pcDNA3.1 group and STAT3-Flag group were further divided into four parts,which were treated with 2 ng/mL TGF-β1 and 0,12.5,25,50 mg/mL CSP for 24 h,respectively.Western blotting was used to detect the protein levels of p-STAT3 Tyr705,COL1A1,vimentin,and α-SMA.Results Forty-five potential anti-liver fibrosis target genes of CSP were obtained,among which 12 were core target genes,with STAT3 having the highest degree.These genes were enriched in cell migration,response to hypoxia,transcription factor binding,protein kinase activity,and other functions,and interleukin signal transduction and the JAK-STAT signal pathway,etc.Com-pared with the Control group,the mRNA and protein levels of COL1A1,vimentin,and α-SMA increased in the TGF-β1 group(all P<0.05).Compared with the TGF-β1 group,the mRNA and protein levels of COL1A1 in the low-,medium-,and high-dose groups decreased,with more significant decreases in the medium-and high-dose groups at the mRNA level(all P<0.05).Compared with the TGF-β1 group,the mRNA and protein levels of vimentin and α-SMA were down-regulat-ed in the high-dose group(all P<0.05).The protein levels of p-STAT3 Tyr705,COL1A1,vimentin,and α-SMA increased successively in the control group,pcDNA3.1 group with TGF-β1,and STAT3-Flag group with TGF-β1(all P<0.05).Compared with the cells in the same group that only received TGF-β1,the relative expression levels of p-STAT3 protein in the pcDNA3.1 group treated with TGF-β1 and low-,medium-,and high-doses of CSP decreased successively(all P<0.05).However,there were no significant changes in the relative expression levels of p-STAT3 protein in the STAT3-Flag group treated with TGF-β1 and low-,medium-,and high-doses of CSP(all P>0.05).The protein levels of COL1A1,vimentin,and α-SMA in the STAT3-Flag group were higher than those in the pcDNA3.1 group(all P<0.05).Conclusion The core target genes of CSP in treating hepatic fibrosis include STAT3,whose mechanism of inhibiting the activation of hepatic stellate cells may be associated with the regulation of the STAT3 signaling pathway.关键词
柴胡疏肝散/网络药理学/肝纤维化/肝星状细胞/信号传导转录激活因子3信号通路Key words
Chaihu Shugan powder/network pharmacology/liver fibrosis/hepatic stellate cells/signal transduc-er and activator of transcription 3 signaling pathway分类
医药卫生引用本文复制引用
王帅,葛茂旭,刘安昌..柴胡疏肝散治疗肝纤维化核心靶基因筛选及对肝星状细胞活化的影响[J].山东医药,2025,65(3):1-6,6.基金项目
山东省自然科学基金青年基金资助项目(ZR2021QH204). （ZR2021QH204）
