摘要
目的:借助转录组高通量测序(RNA-seq)探究调肝理脾方治疗非酒精性脂肪性肝病(NAFLD)的作用机制。方法:将18只小鼠随机分为空白组、模型组、中药组,每组6只,空白组采用常规饲料喂养,另两组采用高脂饮食喂养制备NAFLD模型。造模7周后,中药组给予调肝理脾方颗粒剂灌胃干预,而空白组和模型组则给予等量蒸馏水灌胃。通过苏木精-伊红(HE)染色观察各组小鼠肝组织病理变化,生化分析仪检测血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST))、总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)和高密度脂蛋白胆固醇(HDL-C)的水平。每组随机选取3只小鼠进行肝组织RNA-seq,通过差异基因分析、GO和KEGG富集分析、蛋白互作网络筛选调肝理脾方核心靶点和通路,同时利用数据集GSE89632进行核心靶点外部验证。结果:与空白组比较,模型组小鼠肝组织出现明显的脂肪空泡,中药组小鼠经调肝理脾方治疗后肝组织病理改变明显改善。与模型组比较,中药组小鼠体质量及血清ALT、TC、LDL-C水平均显著改善,血清HDL-C水平升高(P<0.05)。RNA-seq分析结果显示,中药组和模型组存在差异表达基因925个,模型组和空白组有差异表达基因782个。中药组逆转模型组可注释差异表达基因共350个,包括过氧化物酶体增殖物激活受体α(PPARA)、RPL38、RPS20等,参与脂肪酸代谢、脂质代谢、细胞器等过程的调节,还可通过过氧化物酶体增殖物激活受体(PPAR)、核糖体合成等信号通路发挥调控作用。外部数据集验证表明,PPARA、RPL38、RPS20、RPL39在NAFLD疾病进展过程中存在表达量的变化。结论:调肝理脾方能有效改善NAFLD,可能通过调节PPARA、RPL38、RPS20等靶点调控脂质代谢、影响核糖体合成等途径发挥治疗效果。
Objective:To investigate the mechanisms of Tiaogan Lipi formula(TGLPF)in treating non-alcoholic fatty liver disease(NAFLD)through high-throughput transcriptome sequencing.Methods:Eighteen mice were randomly divided into a control group,a model group,and a Chinese medicine group,with six in each group.NAFLD models were prepared by feeding a high-fat diet.After 7 weeks of modeling,the Chinese medicine group received TGLPF via intragastric intervention,while both the control and model groups were administered an equal amount of distilled water.HE staining assessed pathological changes in liver tissue across three groups.A biochemical analyzer measured the levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC),triglycerides(TG),low-density lipoprotein cholesterol(LDL-C),and high-density lipoprotein cholesterol(HDL-C)in each group.Transcriptome sequencing(RNA-seq)was performed on liver tissue from three randomly selected mice in each group.Differential gene analysis,GO and KEGG enrichment analysis,and protein-protein interaction networks were used to identify core targets and pathways of TGLPF.Core targets were externally validated in the GSE89632 dataset.Results:Compared with the control group,the model group mice showed more fat vacuoles in liver tissue,which were significantly improved after treatment with TGLPF.Compared with the model group,TGLPF significantly improved the body weight,ALT,TC,and LDL-C levels in the model group mice,and increased the level of serum HDL-C(P<0.05).RNA-seq analysis found 925 differential genes between the Chinese medicine group and the model group,and 782 differential genes between the model group and control group.A total of 350 annotated differential genes were reversed by the Chinese medicine group compared to the model group,including genes such as PPARA,RPL38,RPS20.Both of these genes are involved in the regulation of fatty acid metabolism,lipid metabolism,and cellular organelles,and may also exert regulatory effects through PPAR and ribosome synthesis signaling pathways.External dataset validation showed that PPARA,RPL38,RPS20,RPL39 exhibited changes in expression during the progression of NAFLD.Conclusion:TGLPF effectively improves NAFLD,possibly by regulating targets such as PPARA,RPL38,RPS20 which influence lipid metabolism and ribosome synthesis,thus providing therapeutic benefits.
作者
屈祥科
董佳
沈悦
孙劲晖
QU Xiang-ke;DONG Jia;SHEN Yue;SUN Jin-hui(Department of Gastroenterology Department of Gastroenterology,Dongzhimen Hospital,Beijing University of Chinese Medicine(Beijing,100700),China)
出处
《中西医结合肝病杂志》
2025年第12期1485-1491,共7页
Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases
关键词
非酒精性脂肪性肝病
调肝理脾方
转录组测序
脂质代谢
Tiaogan Lipi formula
nonalcoholic fatty liver disease
transcriptome sequencing
molecular mechanisms
lipid metabolism
