基于AMPK/PPARα/PGC-1α信号通路探讨金合欢素改善代谢综合征大鼠骨骼肌能量代谢紊乱作用机制  

Study on the Mechanism of Acacetin in Improving Energy Metabolism Disorder in Skeletal Muscle of Rats with Metabolic Syndrome Based on AMPK/PPARα/PGC-1αSignaling Pathway

作　　者：陈祎 袁培培[1,2] 高丽媛 魏亚新 李潘营 阮元 郑晓珂 冯卫生[1,2] CHEN Yi;YUAN Peipei;GAO Liyuan;WEI Yaxin;LI Panying;RUAN Yuan;ZHENG Xiaoke;FENG Weisheng(Henan University of Chinese Medicine,Zhengzhou 450046;The Engineering and Technology Center for Chinese Medicine Development of Henan Province,Zhengzhou 450046)

机构地区：[1]河南中医药大学,郑州450046 [2]河南省中药开发工程技术研究中心,郑州450046

出　　处：《中药药理与临床》2023年第4期29-33,共5页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家重点研发计划-中医药现代化研究项目(编号:2019YFC1708802);河南省重大科技专项(编号:171100310500);河南省高层次人才特殊支持计划(编号:ZYQR201810080);河南中医药大学博士科研基金项目(编号:-RSBSJJ2018-04)。

摘　　要：目的:探究金合欢素对代谢综合征大鼠骨骼肌能量代谢紊乱的影响,并初步探讨其作用机制。方法:通过果糖诱导自发性高血压(SHR)大鼠建立代谢综合征大鼠模型,灌胃给予金合欢素25、50 mg/kg, 7 w后检测收缩压(SBP)、胰岛素抵抗数指数(HOMA-IR),试剂盒检测血清高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白胆固醇(LDL-C)、三酰甘油(TG)的含量;ELISA法检测腓肠肌组织中ATP含量;采用qPCR法测定大鼠腓肠肌中线粒体DNA(mtDNA)拷贝数;Western Blot法检测UCP 3、线粒体融合蛋白(MFN2)、线粒体裂变因子(MFF)、线粒体动力相关蛋白(Drp1)、线粒体途径凋亡指标蛋白(Cytochrome c)、能量代谢相关蛋白AMPK、p-AMPK、PPAR α、PGC-1α的表达。结果:与正常对照组相比,模型对照组大鼠SBP、HOMA-IR、体质量以及血清中LDL-C、TG含量显著升高(P<0.01),血清中HDL-C、腓肠肌质量、ATP含量、mtDNA拷贝数显著降低(P<0.01),腓肠肌组织线粒体中UCP 3、MFN2、Cytochrome c蛋白表达显著下调(P<0.01),MFF、Drp1蛋白表达显著上调(P<0.01),腓肠肌组织中AMPK蛋白磷酸化表达、PPAR α、PGC-1α蛋白表达显著下调(P<0.01);与模型对照组相比,金合欢素各组能明显改善代谢综合征大鼠SBP、HOMA-IR、体质量以及血清中HDL-C、LDL-C、TG的含量;升高腓肠肌质量和ATP含量、mtDNA拷贝数,明显上调UCP 3、MFN2、Cytochrome c蛋白表达,下调MFF、Drp1蛋白表达(P<0.05或P<0.01),促进大鼠腓肠肌线粒体融合并抑制其分裂;上调AMPK蛋白磷酸化、PPAR α、PGC-1α蛋白表达(P<0.01)。结论:金合欢素可能通过激活AMPK/PPAR α/PGC-1α通路,促进代谢综合征大鼠骨骼肌线粒体融合并抑制其分裂,增强线粒体功能进而改善代谢综合征大鼠骨骼肌能量代谢紊乱。Objective:To explore the effects of acacetin on energy metabolism disorder in skeletal muscle of rats with metabolic syndrome and its mechanism.Methods:In this study,the rat model of metabolic syndrome was established by administration of a high-fructose diet to spontaneous hypertension(SHR)rats.Seven weeks after intragastric administration of 25 and 50 mg/kg acacetin,systolic blood pressure(SBP)and homeostasis model assessment-insulin resistance(HOMA-IR)index were detected.The contents of high-density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(LDL-C)were determined by the corresponding colorimetric assay kits(double reagents),and the serum level of triglyceride(TG)was detected by GPO-PAP method.ATP content in gastrocnemius tissue was determined by ELISA.The copy number of mitochondrial DNA(mtDNA)in rat gastrocnemius was determined by real-time quantitative PCR(qPCR).Western Blot was used to detect UCP3,mitochondrial fusion protein mitofusin 2(MFN2),mitochondrial fission factor(MFF),dynamin related protein 1(Drp1),indicator of mitochondrial-mediated apoptosis(cytochrome C),and energy metabolism-related proteins AMPK,p-AMPK,PPARαand PGC-1α.Results:Compared with normal control group,SBP,HOMA-IR,body weight,serum LDL-C and TG levels in model group were significantly increased(P<0.01),while the serum HDL-C level,gastrocnemius weight,ATP content and the copy number of mtDNA in model group were significantly decreased(P<0.01).The expression levels of UCP3,MFN2 and cytochrome C in mitochondria of gastrocnemius tissue were significantly down-regulated(P<0.01),while the expression levels of MFF and Drp1 were significantly up-regulated(P<0.01).The phosphorylation level of AMPK,and the protein expression levels of PPARαand PGC-1αin gastrocnemius tissue were significantly down-regulated(P<0.01).Compared with model group,SBP,HOMA-IR,body weight and serum levels of HDL-C,LDL-C and TG in rats with metabolic syndrome were significantly improved by acacetin(P<0.05 or P<0.01).The gastrocnemius weight

关 键 词：代谢综合征 肌肉 线粒体 腺苷酸活化蛋白激酶/过氧化物酶体增殖激活受体α/过氧化物酶体增殖受体γ辅激活因子α 能量代谢 

分 类 号：R285.5[医药卫生—中药学]