中国中西医结合杂志2024，Vol.44Issue(2)：208-215,8.DOI:10.7661/j.cjim.20230621.280
祛湿活血方作用非酒精性脂肪肝肝细胞转录组分析及验证
Transcriptome Analysis and Verification of the Effect of Qushi Huoxue Recipe on Non-alcoholic Fatty Liver Cells
摘要
Abstract
Objective To obtain and dig out the transcriptome database of Qushi Huoxue Formula acting on nonalcoholic fatty liver disease(NAFLD)hepatocytes and the differentially expressed genes of pyrolysis related to the inflammatory environment of hepatocytes.Methods The research used palmitic acid-induced human liver cell line L-02 to establish the cell model of NAFLD.Transcriptome sequencing with Ribosome RNA removed(Ribo-Zero RNA-Seq)was used to transcriptome sequence the L-02 cell model treated by Qushi Huoxue Recipe.Systematic bioinformatics analysis was performed and differentially expressed genes related to inflammatory environment pyroptosis were screened out.Then the expression of nucleotides bind oligomerized domain-like receptor proteins 3(NLRP3)and its induced liver pyroptosis related molecules were verified by real-time PCR quantitative(qRT-PCR)and Western Blot.Results More than 3 000 significantly differentially expressed genes were obtained in this sequencing analysis.Of them,the number of differentially expressed genes was the highest in the blank treatment group/Qushi Huoxue Formula group,including 1 435 up-regulated and 658 down-regulated genes.Differentially expressed genes were mainly enriched in related pathways such as inflammation signals,cell fate,and fat metabolism.Pyroptosis related genes such as NLRP3,gasdermin D(GSDMD),and cysteinyl aspartate specific proteinase-1(Caspase-1)were screened out.Results of qRT-PCR and Western Blot proved that the treatment of hepatocyte by Qushi Huoxue Formula lowered the expressions of the above molecules at gene and protein levels.Conclusions Qushi Huoxue Formula regulated the gene and protein expressions of inflammation-related pyroptosis signal molecules NLRP3,GSDMD,and Caspase-1,attenuated inflammatory reactions and inhibited hepatocyte pyrolysis.This might be an effective target of Qushi Huoxue Formula in the treatment of NAFLD.关键词
祛湿活血方/转录组测序/肝细胞/非酒精性脂肪肝/差异表达基因/中药复方Key words
Qushi Huoxue Formula/transcriptome sequencing/hepatocyte/non-alcoholic fatty liver/differentially expressed gene/Chinese herbal compound引用本文复制引用
庞华珍,李辉,刘旭东,赵晓芳,唐艳芳..祛湿活血方作用非酒精性脂肪肝肝细胞转录组分析及验证[J].中国中西医结合杂志,2024,44(2):208-215,8.基金项目
国家自然科学基金资助项目(No.82160837) （No.82160837）
2019年广西一流学科建设课题项目(No.2019XK139) （No.2019XK139）
广西中医药大学岐黄工程高层次人才培育项目(No.2021007) （No.2021007）
2019年中医药传承与创新人才培养平台建设项目(No.国中医药人教函[2019]41号) （No.国中医药人教函[2019]41号）
