基于AMPK/SIRT1/PGC-1α信号通路研究香青兰总黄酮对大鼠心肌缺血再灌注损伤的保护机制  

Study on Protective Mechanism of Dracocephalum moldavica Total Flavonoids against Myocardial Ischemia-reperfusion Injury in Rats Based on AMPK/SIRT1/PGC-1αSignaling Pathway

作　　者：赵云丽 袁勇[1,2] 马晓莉[1,2] 黄川生 文志萍[1,2] 郭新红[1,2] 王新春 ZHAO Yunli;YUAN Yong;MA Xiaoli;HUANG Chuansheng;WEN Zhiping;GUO Xinhong;WANG Xinchun(College of Pharmacy,Shihezi University,Xinjiang Shihezi 832002,China;Dept.of Pharmacy,the First Affiliated Hospital of the Medical College,Shihezi University,Xinjiang Shihezi 832008,China)

机构地区：[1]石河子大学药学院,新疆石河子832002 [2]石河子大学医学院第一附属医院药剂科,新疆石河子832008

出　　处：《中国药房》2021年第3期278-283,共6页China Pharmacy

基　　金：国家自然科学基金资助项目(No.81860747,No.81960766)。

摘　　要：目的:研究香青兰总黄酮(TFDM)对腺苷酸活化蛋白激酶(AMPK)/沉默信息调节因子1(SIRT1)/过氧化物酶体增殖物激活受体γ辅激活因子1α(PGC-1α)信号通路的影响,探究其保护大鼠心肌缺血再灌注损伤(MIRI)的作用机制。方法:将50只健康雄性SD大鼠随机分为假手术组、模型组、TFDM组[60 mg/(kg·d),以提取物计]、Compound C+TFDM组[灌胃60 mg/(kg·d)TFDM+再灌注前15 min尾静脉注射250μg/kg Compound C(AMPK抑制剂)]、EX-527+TFDM组[灌胃60 mg/(kg·d)TFDM+再灌注前20 min腹腔注射5 mg/kg EX-527(SIRT1抑制剂)],每组10只。每天灌胃给药1次,连续7 d。末次灌胃给药后,假手术组大鼠行假手术,其余4组大鼠均采用结扎冠状动脉左前降支缺血30 min、再灌注2 h构建MIRI模型。再灌注结束后,采用苏木精-伊红染色法观察大鼠心肌组织病理学变化;采用反相高效液相色谱法测定其心肌组织中腺苷三磷酸(ATP)、腺苷二磷酸(ADP)、腺苷一磷酸(AMP)及烟酰胺腺嘌呤二核苷酸(NAD^+)的含量;采用实时荧光定量-聚合酶链式反应法检测大鼠心肌组织中AMPK、SIRT1和PGC-1αmRNA表达水平;采用Western blotting法检测大鼠心肌组织中AMPK蛋白的磷酸化水平和SIRT1、PGC-1α蛋白表达水平。结果:与假手术组比较,模型组大鼠心肌纤维排列紊乱、横向条纹消失,细胞肿胀破裂、坏死,细胞核变形移位;心肌组织中ATP、NAD^+含量和AMPK、SIRT1、PGC-1αmRNA表达水平以及SIRT1、PGC-1α蛋白表达水平均显著降低(P<0.05或P<0.01),ADP、AMP含量及AMPK蛋白的磷酸化水平均显著升高(P<0.01)。与模型组比较,TFDM组大鼠心肌病理学形态明显改善;心肌组织中ATP、NAD^+含量和AMPK、SIRT1、PGC-1αmRNA表达水平以及AMPK蛋白的磷酸化水平和SIRT1、PGC-1α蛋白表达水平均显著升高(P<0.05或P<0.01),ADP、AMP含量均显著降低(P<0.01)。与TFDM组比较,Compound C+TFDM组和EX-527+TFDM组大鼠上述指标的改善作用均被逆转(P<0.05或POBJECTIVE:To study the effects of Dracocephalum moldavica total flavonoids(TFDM)on AMPK/SIRT1/PGC-1αsignaling pathway,and to explore the mechanism of its protective effect on myocardial ischemia reperfusion injury(MIRI)rats.METHODS:Totally 50 healthy male SD rats were randomly divided into sham operation group,model group,TFDM group[60 mg/(kg·d),by extract],Compound C+TFDM group[ig administration of 60 mg/(kg·d)TFDM+intravenous injection of 250μg/kg Compound C(AMPK inhibitor)via tail vein 15 min before reperfusion],EX-527+TFDM group[ig administration of 60 mg/(kg·d)TFDM+ip injection of 5 mg/kg EX-527(SIRT1 inhibitor)20 min before reperfusion],with 10 rats in each group.They were given relevant medicine intragastrically,once a day,for consecutive 7 days.After last ig administration,sham operation group underwent sham operation,other 4 groups were established MIRI model by ligating left anterior descending coronary artery,ischemia for 30 min and reperfusion for 2 h.After reperfusion,the myocardial histopathological changes were observed by HE staining;RP-HPLC method was used to determine the contents of ATP,ADP,AMP and NAD^+in cardiac tissue.mRNA expressions of AMPK,SIRT1 and PGC-1αwere detected by quantitative real-time PCR assay.Western blotting assay was used to detect the phosphorylation level of AMPK protein and the expressions of SIRT1 and PGC-1αprotein in myocardium.RESULTS:Compared with sham operation group,model group showed myocardial fibers arranged disorder and horizontal stripes disappearance,cell swelling burst and necrosis,and nuclei deformation displacement;the contents of ATP and NAD^+,mRNA expression of AMPK,SIRT1 and PGC-1α,protein expression of SIRT1 and PGC-1αin cardiac tissue were decreased significantly(P<0.05 or P<0.01);the contents of ADP and AMP,the phosphorylation level of AMPK protein were increased significantly(P<0.01).Compared with model group,myocardial pathological morphology were improved significantly in TFDM group;the contents of ATP and NAD^+in cardiac tissue,mRNA expre

关 键 词：香青兰总黄酮 能量代谢 腺苷酸活化蛋白激酶 沉默信息调节因子1 过氧化物酶体增殖物激活受体γ辅激活因子1α 机制 

分 类 号：R285[医药卫生—中药学]