基于调控SIRT3/PGC-1α通路干预能量代谢探讨温胆汤防治心肌缺血机制  

Wendantang Regulates Energy Metabolism in Treatment of Myocardial Ischemia via SIRT3/PGC-1αPathway

作　　者：张鑫俊 肖志强 鲁佳[2] 顿文亮[2] 顾宁[2] ZHANG Xinjun;XIAO Zhiqiang;LU Jia;DUN Wenliang;GU Ning(Nanjing University of Chinese Medicine,Nanjing 210046,China;Nanjing Hospital of Chinese Medicine Affiliated to Nanjing University of Chinese Medicine,Nanjing 210022,China)

机构地区：[1]南京中医药大学,南京210046 [2]南京中医药大学附属南京中医院,南京210022

出　　处：《中国实验方剂学杂志》2025年第2期1-8,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：南京市医学重点科技发展项目(ZKX21060);江苏省第四批名老中医药专家传承工作室建设项目(苏中医科教〔2021〕7号);江苏省研究生实践创新计划项目(SJCX24_1069)。

摘　　要：目的:探讨温胆汤调控沉默信息调节因子3(SIRT3)/过氧化物酶体增殖物激活受体γ共激活-1α(PGC-1α)通路影响能量代谢从而防治高脂血症(HL)伴心肌缺血(MI)模型大鼠心肌缺血的机制。方法:30只SD大鼠随机分为空白组、模型组、温胆汤低剂量组、温胆汤高剂量组、曲美他嗪组,每组6只。空白组常规饲养,其余各组采用连续6周高脂饲料喂养,构建大鼠HL模型。随后,温胆汤低、高剂量组分别以3.702、7.404 g·kg^(-1)·d^(-1)剂量灌胃,曲美他嗪组以0.006 g·kg^(-1)·d^(-1)剂量灌胃,空白组和模型组以等量生理盐水灌胃,连续灌胃14 d,末次灌胃后1 h,除空白组,其余各组大鼠腹腔注射垂体后叶素(30 U·kg^(-1)),制备大鼠MI模型。心电图(ECG)检测大鼠心电图变化;苏木素-伊红(HE)染色观察心脏病理学改变;酶联免疫吸附测定法(ELISA)检测血清心肌肌钙蛋白(I cTnI)、肌红蛋白(MYO)、肌酸激酶同工酶(CK-MB)水平;比色法检测血清总胆固醇(TC)、甘油三酯(TG)及心肌组织三磷酸腺苷(ATP)、丙二醛(MDA)、超氧化物歧化酶(SOD)水平;蛋白免疫印迹法(Western blot)检测大鼠心肌组织SIRT3、PGC-1α、腺苷酸活化蛋白激酶(AMPK)及磷酸化(p)-AMPK蛋白表达水平;实时荧光定量聚合酶链式反应(Real-time PCR)检测大鼠心肌组织SIRT3、PGC-1α、AMPK mRNA表达。结果:与空白组比较,模型组大鼠心电图J点偏移和升高,心率变快;心肌组织心肌横纹排列紊乱,边界不清晰;CK-MB、cTnI、MYO升高(P<0.05,P<0.01),TC、TG升高(P<0.01),SOD、ATP下降(P<0.01);SIRT3、PGC-1α、AMPK mRNA表达下降(P<0.05),SIRT3、PGC-1α、p-AMPK蛋白水平下调(P<0.05)。与模型组比较,温胆汤低、高剂量组与曲美他嗪组有抑制心电图J点偏移和升高、减慢心率的作用,心肌组织炎性细胞浸润与心肌横纹排列紊乱减轻,CK-MB、cTnI、MYO下降(P<0.05,P<0.01),TC、TG下降(P<0.05,P<0.01),SOD升高(P<0.01);SIRT3、PGC-1α、AMPK mRNA表达�Objective:To investigate the mechanism by which Wendantang regulates the silent information regulator 3(SIRT3)/peroxisome proliferator-activated receptor-gamma coactivator-1α(PGC-1α)pathway to influence energy metabolism and thereby prevent and treat myocardial ischemia(MI)in a rat model of hyperlipidemia(HL).Methods:Thirty SD rats were randomly assigned into five groups:control,model,low-dose(3.702 g·kg^(-1)·d^(-1))Wendantang,high-dose(7.404 g·kg^(-1)·d^(-1))Wendantang,and positive control(trimetazidine,0.006 g·kg^(-1)·d^(-1)),with six rats in each group.The control group was fed normally,while the other groups were fed with a high-fat diet for six weeks for the modeling of HL.Subsequently,the drug intervention groups were administrated with corresponding drugs by gavage,and the control and model groups received an equivalent volume of normal saline for 14 days.One hour after the last gavage,the other groups except the control group were injected intraperitoneally with posterior pituitary hormone(30 U·kg^(-1))to induce MI.Electrocardiography(ECG)was employed to detect changes in the electrocardiogram.Hematoxylin-eosin staining was performed to observe cardiac pathological changes.Enzyme-linked immunosorbent assay was employed to measure the serum levels of cardiac troponin(I cTnI),myoglobin(MYO),and creatine kinase-MB(CK-MB).Colorimetry was used to determine the levels of total cholesterol(TC)and triglycerides(TG)in the serum and ATP,malondialdehyde(MDA),and superoxide dismutase(SOD)in the myocardial tissue.Western blot was employed to determine the protein levels of SIRT3,PGC-1α,adenosine monophosphate-activated protein kinase(AMPK),and phosphorylated AMPK(p-AMPK)in the myocardial tissue.Real-time PCR was employed to measure the mRNA levels of SIRT3,PGC-1α,and AMPKαin the myocardial tissue.Results:Compared with the control group,the model group showed significant J-point deviation and elevation in the ECG image,increased heart rate,disarrangement of myocardial fibers with unclear boundaries,elevated

关 键 词：温胆汤 心肌缺血 高脂血症 氧化应激 能量代谢 沉默信息调节因子3(SIRT3)/过氧化物酶体增殖物激活受体γ共激活因子-1α(PGC-1α)信号通路 

分 类 号：R285[医药卫生—中药学] R289[医药卫生—中医学] R256