非酒精性脂肪性肝炎代谢相关关键差异表达基因靶向治疗药物的筛选与验证OACSTPCD
Screening and validation of targeted therapeutic agents for key differentially expressed genes related to metabolism in non-alcoholic steatohepatitis
目的 筛选非酒精性脂肪性肝炎(NASH)代谢相关关键差异表达基因的靶向治疗药物,并进行验证,为NASH的新药研发提供参考.方法 从基因表达数据库(GEO)中下载NASH基因表达谱GSE164760和GSE63067数据集,利用R软件筛选差异表达基因(DEGs),并进行GO分子功能与KEGG信号通路分析.将NASH DEGs与代谢相关基因取交集,采用LASSO回归模型和SVM-RFE算法筛选NASH代谢相关关键DEGs.利用比较毒物基因组学数据库(CTD)、中药系统药理学数据库与分析平台(TCMSP)、Pubmed数据库筛选NASH代谢相关关键差异表达基因的治疗药物.利用Autodock4软件将获得的治疗药物与NASH代谢相关关键DEGs进行分子对接验证靶向关系.采用油红〇实验观察NASH代谢相关关键差异表达基因靶向治疗药物柚皮素对NASH细胞内脂滴含量的影响观察,采用实时荧光定量RT-qPCR法观察NASH代谢相关关键差异表达基因靶向治疗药物柚皮素对NASH细胞内NASH代谢相关关键DEGs的影响.结果 筛选得到336个DEGs,主要参与脂肪酸代谢过程、蛋白质导向和小分子分解代谢过程和辅助因子的生物合成、胆汁分泌和过氧物酶体等信号通路.NASH代谢相关关键DEGs为SLCO1B1、SCD、ACAT1基因.共同靶向ACAT1、SCD、SLCO1B1的化合物有4种,分别为柚皮素、环孢霉素、红豆碱、鹅去氧胆酸.分子对接验证结果显示,柚皮素、环孢霉素、红豆碱、鹅去氧胆酸与ACAT1、SCD、SLCO1B1基因之间具有较好的分子结合活性,具有靶向关系.细胞实验验证结果显示,靶向治疗药物柚皮素可降低NASH细胞内脂滴含量,影响NASH细胞内ACAT1、SCD、SLCO1B1基因表达.结论 筛选获得的NASH代谢相关关键差异表达基因靶向治疗药物是柚皮素、环孢霉素、红豆碱、鹅去氧胆酸,分子对接验证和细胞实验均验证通过.
Objective To screen and validate targeted therapeutic agents for key differentially expressed genes(DEGs)related to metabolism in non-alcoholic steatohepatitis(NASH),in order to provide a reference for the develop-ment of new drugs for NASH.Methods The NASH gene expression profiles GSE164760 and GSE63067 datasets were downloaded from the Gene Expression Omnibus(GEO)database.Differentially expressed genes(DEGs)were identified using R software,and GO molecular function and KEGG signaling pathway analyses were performed.The DEGs associat-ed with NASH and metabolic genes were intersected to screen NASH metabolic key DEGs using LASSO regression model and SVM-RFE algorithm.Targeted therapeutic agents for NASH metabolic key DEGs were identified using the Compara-tive Toxicogenomics Database(CTD),Traditional Chinese Medicine Systems Pharmacology Database and Analysis Plat-form(TCMSP),and PubMed database.Molecular docking was conducted using Autodock4 software to verify the targeted relationship between the obtained therapeutic agents and NASH metabolic key DEGs.The effect of naringenin,a NASH metabolic key DEGs targeted therapeutic agent,on the content of intracellular lipid droplets in NASH cells was observed using Oil Red 〇staining.The effect of naringenin on the expression of NASH metabolic key DEGs within NASH cells was observed using real-time quantitative RT-qPCR.Results A total of 336 DEGs were identified,which were mainly involved in fatty acid metabolism,protein targeting,small molecule degradation metabolism,the biosynthesis of auxiliary factors,bile secretion,and peroxisomes,and other signaling pathways.The NASH metabolic key DEGs were identified as SLCO1B1,SCD,and ACAT1 genes.Four compounds,naringenin,cyclosporin,abrine,and chenodeoxycholic acid,were found to target ACAT1,SCD,and SLCO1B1.Molecular docking results indicated that naringenin,cyclosporin,abrine,and chenodeoxycholic acid had good molecular binding activity with ACAT1,SCD,and SLCO1B1 genes,sug-gesting a targeted relationship.Cell experiments demonstrated that the targeted therapeutic agent naringenin could de-crease the content of intracellular lipid droplets in NASH cells and influence the expression of ACAT1,SCD,and SL-CO1B1 genes within NASH cells.Conclusion The targeted therapeutic agents for key DEGs related to metabolism in NASH are naringenin,cyclosporin,abrine,and chenodeoxycholic acid,which are validated by molecular docking and cell experiments.
黄雨晴;王小宁;陆景坤
内蒙古医科大学基础医学院,呼和浩特 010010
药学
慢性肝病非酒精性脂肪性肝病非酒精性脂肪性肝炎柚皮素靶向治疗药物
chronic liver diseasenon-alcoholic fatty liver diseasenon-alcoholic steatohepatitisnaringenintar-geted therapeutic agents
《山东医药》 2024 (033)
25-29 / 5
国家自然科学基金项目(81960757,82260813);内蒙古自治区成果转化项目(2019CG042);内蒙古自治区自然科学基金项目(2019MS08119,2020MS08045);内蒙古医科大学基础项目(YKD2021MS033);内蒙古自治区科技计划项目(2023YFDZ0059).
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