芹菜素对大鼠缺血/再灌注心肌细胞凋亡及相关蛋白Bcl-2、Bax、Caspase-3表达的影响

摘要

Aim To investigate the flavonoid apigenin on myocardial cell apoptosis and Bcl-2 , Bax , Caspase-3protein expression in experimental rats with myocardialischemia and reperfusion ( ischemia/reperfusion , I/R )and to alalyze the pathological damage in myocardialtissue. Methods The myocardialischemia-reperfusioninjury model was established through occluding the leftanterior descending branch of coronary artery for 45min and removing the ligation later to reperfuse for 2hours. Sixty-four rats were randomly divided into eightgroups: normal ( Normal ) group, sham operation( Sham ) group, saline ischemia-reperfusion ( NSgroup, solvent control( Sol ) group, metoprolol( Meto )control group , apigenin low, medium and high dose ( 1mg · kg-1, 2 mg · kg-1, 4 mg · kg-1 ) treatmentgroup ( Apil , Api2, Api4 ). Hearts were quickly re-moved after 2 h reperfusion, and myocardial cell apop-tosis was marked by TUNEL; Bcl-2、Bax and Caspase-3 protein expression were tested by immunohistochemi-cal staining method; myocardial injury degree was ob-served though histopathological biopsy. Results Myo-cardial cell apoptosis rate in apigenin groups with dif-ferent doses were significantly lower than that of NSgroup ( P ＜ 0. 05 ). and dose-dependent reduction inmyocardial ischemia/reperfusion was observed; Apige-nin increased dose-dependently the Bcl-2 protein ex-pression( P ＜0. 05 ) and reduced dose-dependently theBax and Caspase-3 protein expression( P ＜ 0. 05 ) inrats with myocardial ischemia; Bathological damage ofmyocardial cells in apigenin groups with different doseswere significantly lighter than that of NS group( P ＜0. 05 ), and dose-dependent reduction was detected inmyocardial ischemia/reperfusion. Conclusion Theprotective effect of apigenin on myocardial ischemia/reperfusion may be related to the inhibition of ischemi-a/reperfusion myocardial apoptosis; The mechanism ofapigenin against myocardial apoptosis may involve theincreased Bcl-2 protein expression and reduced Bax.Caspase-3 protein expression; Apigenin can reducepathological damage of myocardial cells.%目的 探讨黄酮类化合物芹菜素对大鼠实验性心肌缺血/再灌注(ischemia/reperfusion,I/R)时心肌细胞凋亡与Bcl-2、Bax、Caspase-3蛋白表达的影响,并分析心肌组织病理学损伤程度.方法 采用结扎左冠状动脉前降支,心肌缺血45 min,再灌注2 h制作缺血/再灌注模型.将大鼠随机分为8组,即正常组(normal group,Normal)、假手术组(sham operation group,Sham)、生理盐水缺血/再灌注组(saline ischemia-reperfusion group,NS)、溶剂对照组(solvent control group,Sol)、美托洛尔对照组(metoprolol control group,Meto)、芹菜素低、中、高剂量(1、2、4 mg·kg-1)用药组(apigenin low,medium and high dose treatment group,Api1,Api2,Api4).再灌注2 h后迅速取出心脏,TUNEL法原位标记凋亡的心肌细胞;免疫组化法测Bcl-2、Bax和Caspase-3蛋白表达;做病理组织切片检查心肌损伤情况.结果 芹菜素各剂量组心肌细胞凋亡率明显低于NS组(P＜0.05),Api1,Api2,Api4能剂量依赖性地降低大鼠缺血/再灌注心肌细胞凋亡率;芹菜素各剂量组剂量依赖性地提高大鼠心肌缺血/再灌注的Bcl-2 蛋白表达量(P＜0.05)、降低大鼠心肌缺血/再灌注的Bax、Caspase-3 蛋白表达量(P＜0.05);芹菜素各剂量组与NS组比较,心肌组织损伤的病理学变化明显减轻(P＜0.05).结论 芹菜素对缺血/再灌注心肌的保护效应可能与其抑制缺血/再灌注心肌细胞凋亡有关;芹菜素抗心肌凋亡作用的机制可能与其上调Bcl-2蛋白表达和下调Bax、Caspase-3蛋白表达有关;芹菜素能明显减轻心肌组织损伤.