左归降糖清肝方含药血清调控线粒体功能改善HepG2细胞脂肪性变的机制研究  

Study on the mechanism of serum containing Zuogui Jiangtang Qinggan Formula(左归降糖清肝方)in regulating mitochondrial function to ameliorate steatosis in HepG2 cells

作　　者：李亚楠 冯馨瑶 徐睿怡 李振祥 马桂秀 李萍 成细华[2] LI Ya-nan;FENG Xin-yao;XU Rui-yi;LI Zhen-xiang;MA Gui-xiu;KI Ping;CHENG Xi-hua(Graduate School of Hunan University of Chinese Medicine,Changsha 410208,China;Medical School of Hunan University of Chinese Medicine,Changsha 410208,China;Xiangya Boai Rehabilitation Hospital,Changsha 410100,China)

机构地区：[1]湖南中医药大学研究生院,湖南长沙410208 [2]湖南中医药大学医学院湖南长沙410208 [3]湘雅博爱康复医院,湖南湘雅410100

出　　处：《时珍国医国药》2025年第6期1014-1022,共9页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学区域联合基金(U21A20411);湖南省卫生健康委员会重点项目(A202303066904);湖南省自然科学基金(2025JJ90275,2025JJ90309);湖南中医药大学研究生创新课题(2023CX159);湖南中医药大学本科生科研创新基金(2023BKS068,2024BKS093);湖南中医药大学一流学科基础医学资助项目;湖南省国内一流培育学科中西医结合学科(2024);血管生物学与转化医学湖南省重点实验室(2024);中西医结合病原生物学湖南省重点实验室(2024)。

摘　　要：目的探讨左归降糖清肝方含药血清调控过氧化物酶体增殖物激活受体-γ共激活因子-1α(proliferator-activated receptor-γcoctiovator-1α,PGC-1α)介导的线粒体功能改善HepG2细胞脂肪性变。方法采用油酸(OA)诱导HepG2细胞建立脂肪性变细胞模型,以左归降糖清肝方含药血清进行干预。采用油红O染色检测脂肪堆积;试剂盒检测细胞内甘油三酯(TG)、活性氧(ROS)、ATP、线粒体膜电位;RT-PCR检测PGC-1α及其相关基因mRNA表达;Western blotting检测PGC-1α及其相关蛋白表达。结果与空白组比较,模型组TG及ROS含量显著升高(P<0.01);ATP含量、线粒体膜电位显著下降(P<0.01);PGC-1α、NRF1、TFAM及下游脂代谢相关基因PPARα、ACOX1、CPT1αmRNA表达显著下调(P<0.05),脂肪合成基因FAS mRNA表达显著上调(P<0.05);与模型组比较,左归降糖清肝方含药血清各剂量组均可改善细胞内脂质沉积,显著下降TG、ROS含量(P<0.01);显著上调ATP、线粒体膜电位含量(P<0.01);显著上调PGC-1α、NRF1、TFAM及下游PPARα、ACOX1、CPT1αmRNA表达(P<0.05),显著下调FAS mRNA表达(P<0.05);PGC-1α抑制剂降低了左归降糖清肝方的保护作用(P<0.05)。结论左归降糖清肝方含药血清通过调控PGC-1α表达,影响PGC-1α调控的线粒体生物发生和脂质代谢,改善线粒体功能,减少肝细胞脂肪性变。Objective To investigate the mechanism by which the medicated serum of Zuogui Jiangtang Qinggan Formula(左归降糖清肝方,ZGJTQGF)regulates peroxisome proliferator-activated receptor-γcoactivator-1α(PGC-1α)-mediated mitochondrial function to ameliorate hepatic steatosis in HepG2 cells.Methods A cellular model of hepatic steatosis was established using oleic acid(OA)-induced HepG2 cells,followed by intervention with serum containing ZGJTQGF.Lipid accumulation was detected via Oil Red O staining.Intracellular triglyceride(TG),reactive oxygen species(ROS),ATP levels,and mitochondrial membrane potential(MMP)were measured using the assay kits.RT-PCR and Western blot(WB)were used to evaluate mRNA and protein expression of PGC-1αand its related genes.Results Compared with the control group,the model group showed significantly increased TG and ROS levels(P<0.01),decreased ATP content and MMP(P<0.01),down-regulated mRNA expressions of PGC-1α,NRF1,TFAM,and downstream lipid metabolism-related genes(PPARα,ACOX1,CPT1α)(P<0.05),and up-regulated mRNA expression of the lipogenic gene FAS(P<0.05).Compared with the model group,all dose groups with medicated serum of ZGJTQGF ameliorated intracellular lipid deposition,significantly reduced TG and ROS levels(P<0.01),increased ATP content and MMP(P<0.01),up-regulated mRNA expressions of PGC-1α,NRF1,TFAM,PPARα,ACOX1,and CPT1α(P<0.05),and down-regulated FAS mRNA expression(P<0.05).The protective effects of ZGJTQGF were diminished by a PGC-1αinhibitor(P<0.05).Conclusion The medicated serum of ZGJTQGF ameliorates hepatic steatosis by regulating the PGC-1αexpression,affecting mitochondrial biogenesis and lipid metabolism while improving mitochondrial function.

关 键 词：非酒精性脂肪肝 左归降糖清肝方 HEPG2细胞 PGC-1Α 线粒体功能 

分 类 号：R285.5[医药卫生—中药学]