摘要
目的探究褪黑素(Mel)介导腺苷酸活化蛋白激酶(AMPK)通路对心肌缺血/再灌注损伤(MI/RI)的保护机制。方法选取50只无特定病原体(SPF)级雄性SD大鼠作为研究对象,随机分为5组,每组10只,其中假手术组(Sham组)不结扎冠状动脉左前降支;Mel组不结扎冠状动脉左前降支,腹腔单次注射褪黑素;溶剂(V)对照组(MI/RI+V组)采用冠状动脉结扎法造模,造模前及再灌注前腹腔注射乙醇-生理盐水;Mel治疗组(MI/RI+Mel组)造模前及再灌注前腹腔注射褪黑素;Mel+AMPK抑制剂BML275处理组(MI/RI+Mel+BML275组)造模前及再灌注前腹腔注射BML275,再灌注前腹腔注射褪黑素。采用冠状动脉结扎法建立MI/RI模型,苏木素-伊红(HE)染色法观察病理结果,检测大鼠心功能和血清肌钙蛋白T(TnT)、乳酸脱氢酶(LDH)、肌酸激酶(CK)水平。Evans blue-2,3,5-氯化三苯基四氮唑(TTC)双染法观察心肌梗死情况。Western blot检测心肌组织腺苷酸活化蛋白激酶α(AMPKα)、线粒体动力相关蛋白(Drp1)和细胞色素C(Cyt C)表达。结果MI/RI+Mel组、MI/RI+Mel+BML275组的左心室射血分数(LVEF)和左心室缩短分数(LVFS)均高于MI/RI+V组,MI/RI+Mel+BML275组的LVEF和LVFS均低于MI/RI+Mel组,差异有统计学意义(P<0.05)。MI/RI+Mel组、MI/RI+Mel+BML275组的血清TnT、CK、LDH水平均低于MI/RI+V组,MI/RI+Mel+BML275组的血清TnT、CK、LDH水平均高于MI/RI+Mel组,差异有统计学意义(P<0.05)。MI/RI+Mel组、MI/RI+Mel+BML275组的心肌梗死面积均小于MI/RI+V组,MI/RI+Mel+BML275组的心肌梗死面积大于MI/RI+Mel组,差异有统计学意义(P<0.05)。MI/RI+Mel组、MI/RI+Mel+BML275组的磷酸化腺苷酸活化蛋白激酶α(p-AMPKα)表达均高于MI/RI+V组,MI/RI+Mel+BML275组的p-AMPKα表达低于MI/RI+Mel组,差异有统计学意义(P<0.05)。MI/RI+Mel组、MI/RI+Mel+BML275组的磷酸化线粒体动力相关蛋白(p-Drp1)/Drp1和Cyt C表达均低于MI/RI+V组,MI/RI+Mel+BML275组的p-Drp1/Drp1和Cyt C表达均高于MI/RI+Mel组,差异有统计学意义(P<0.05)。结论Mel可以通过激活AMPK通路抑制线粒体损伤,并减轻MI/RI损伤,改善心功能。
Objective To explore the protective mechanism of melatonin(Mel)mediated adenosine 5'-monophosphate-activated protein kinase(AMPK)pathway against myocardial ischemia/reperfusion injury(MI/RI).Methods A total of 50 SPF grade male SD rats were selected as research subjects,and they were randomly divided into 5 groups,with 10 rats in each group.In Sham operation group,the left anterior descending coronary artery was not ligated.In the Mel group,the left anterior descending coronary artery was not ligated,and a single intraperitoneal injection of melatonin was given.The vehicle(V)control group(MI/RI+V group)was established by coronary artery ligation,and ethanol-saline was injected intraperitoneally before modeling and reperfusion.In the Mel treatment group(MI/RI+Mel group),melatonin was injected intraperitoneally before modeling and reperfusion.In MI/RI+Mel+BML275 group,BML275 was intraperitoneally injected before modeling and reperfusion,and melatonin was intraperitoneally injected before reperfusion.A MI/RI model was established by coronary artery ligation,and the pathological results were observed by hematoxylin-eosin(HE)staining.The cardiac function and serum levels of troponin T(TnT),lactate dehydrogenase(LDH),and creatine kinase(CK)were detected in rats.Evans blue-triphenyltetrazolium chloride(TTC)double staining method was used to observe myocardial infarction.Western blot was used to detect the expression of adenosine 5'-monophosphate-activated protein kinase(AMPKα),dynamin-related protein 1(Drp1),and cytochrome C(Cyt C)in myocardial tissue.Results The left ventricular ejection fraction(LVEF)and left ventricular shortening fraction(LVFS)of the MI/RI+Mel group and the MI/RI+Mel+BML275 group were higher than those of the MI/RI+V group,while the LVEF and LVFS of the MI/RI+Mel+BML275 group were lower than those of the MI/RI+Mel group,with statistically significant differences(P<0.05).The serum TnT,CK,and LDH levels in the MI/RI+Mel group and MI/RI+Mel+BML275 group were lower than those in the MI/RI+V group,while the serum TnT,CK,and LDH levels in the MI/RI+Mel+BML275 group were higher than those in the MI/RI+Mel group,with statistically significant differences(P<0.05).The myocardial infarction area in the MI/RI+Mel group and MI/RI+Mel+BML275 group were smaller than that in the MI/RI+V group,while the myocardial infarction area in the MI/RI+Mel+BML275 group was larger than that in the MI/RI+Mel group,with statistically significant differences(P<0.05).The expression of phosphorylation adenosine 5'-monophosphate-activated protein kinaseα(p-AMPKα)in the MI/RI+Mel group and the MI/RI+Mel+BML275 group were higher than that in the MI/RI+V group,while the expression of p-AMPKαin the MI/RI+Mel+BML275 group was lower than that in the MI/RI+Mel group,with statistically significant differences(P<0.05).The expression of phosphorylation dynamin-related protein 1(p-Drp1)/Drp1 and Cyt C in the MI/RI+Mel group and MI/RI+Mel+BML275 group were lower than that in the MI/RI+V group,and the expression of p-Drp1/Drp1 and Cyt C in the MI/RI+Mel+BML275 group were higher than that in the MI/RI+Mel group,and the differences were statistically significant(P<0.05).Conclusion Mel can inhibit mitochondrial damage by activating AMPK pathway,reduce MI/RI injury,and improve cardiac function.
作者
曾明辉
余龙辉
邱明涛
李晨
ZENG Minghui;YU Longhui;QIU Mingtao;LI Chen(The Second Department of Cardiovascular Medicine,Pingxiang People's Hospital,Jiangxi Province,Pingxiang337000,China;Department of Cardiovascular Medicine,Jiangxi Provincial People's Hospital,Jiangxi Province,Nanchang330000,China)
出处
《中国当代医药》
2025年第6期10-15,共6页
China Modern Medicine
基金
江西省卫生健康委科技计划项目(202410765)。
关键词
褪黑素
心肌缺血/再灌注损伤
大鼠
腺苷酸活化蛋白激酶通路
线粒体损伤
Melatonin
Myocardial ischemia/reperfusion injury
Adenosine 5’-monophosphate-activated protein kinase pathway
Mitochondrial damage
