环孢素A对心肌缺血再灌注Fas/FasL蛋白表达及心肌细胞凋亡的影响

Impact of Fas/FasL protein expression and myocardial apoptosis of cyclosporine A on myocardial ischemia-reperfusion

目的探讨环孢素A（CSA）对心肌缺血再灌注大鼠Fas/FasL蛋白表达的影响及CSA对心肌细胞的保护作用。方法选取30只SD大鼠随机分为假性手术组（S组）、缺血在灌注组（IR组）、CSA预处理组（CSA组）,每组各10只,CSA组给予每天10mg/kg的CSA与腹腔内注射,10d后建立大鼠心肌缺血再灌注模型,IR组及CSA组结扎左冠状动脉前降肢30min后复灌注3h,S组仅行穿线处理,不进行结扎。分别于结扎前（T0）、再灌注即刻（T1）、30min（T2）、60min（T3）、90min（T4）、120min（T5）采用ELISA测定各组大鼠血清超敏肌钙蛋白I（cTnI）、肌酸激酶同工酶（CK-MB）水平。于试验结束后采用流式细胞仪测定各组大鼠血清心肌细胞凋亡率、钙调神经磷酸酶（CaN）活性,采用免疫组织化学法测定Fas/FasL蛋白水平。结果与T0,IR组、CSA组T1~T5时段cTnI、CK-MB水平显著升高（P〈0.05）。与S组相比,IR组、CSA组T1~T5时段cTnI、CK-MB水平显著升高（P〈0.05）。CSA组T1~T5时段cTnI、CK-MB水平显著低于IR组（P〈0.05）。与IR组,CSA组心肌缺血再灌注后心肌细胞凋亡率、CaN活性及Fas/FasL蛋白水平显著下降（P〈0.05）。结论 CSA预处理心肌缺血再灌注大鼠可通过抑制CaN信号降低大鼠心肌梗死面积,同时能有效减少Fas/FasL蛋白表达,延缓心肌细胞缺血再灌注损伤,具有保护心肌细胞作用。

Objective To investigate the impact of Fas/FasL protein expression and myocardial apoptosis of cyclosporine A on myocardial ischemia-reperfusion.Method 30 SD rats were randomly divided into pseudo-operation group（S group）,ischemic perfusion group（IR）,CSA preconditioning group（CSA group）,each group 10,CSA group was given 10 mg daily/kg intraperitoneal injection of CSA and establish 10 dafter myocardial ischemia-reperfusion model,IR group and CSA group before and after re-perfusion 3hleft anterior descending coronary artery ligation limb 30 min,S group only threading processing,without ligation.Respectively,before ligation（T0）,immediately reperfusion（T1）,reperfusion 30min（T2）,reperfusion 60min（T3）,reperfusion 90min（T4）,reperfusion 120min（T5）using ELISA serum of the rats sensitive troponin I（cTnI）,creatine kinase（CK-MB）levels.After the end of the test serum by flow cytometry rat myocardial apoptosis rate phosphatase calcineurin（CaN）activity was measured Fas/FasL protein level by immunohistochemistry method.Result Compared with T0,IR group,CSA group T1~ T5 period cTnI,CK-MB levels were significantly higher（P〈0.05）.Compared with the S group,IR group,CSA group T1~T5period cTnI,CK-MB levels were significantly higher（P〈0.05）.CSA Group T1~ T5 period cTnI,CK-MB levels were significantly lower than the IR group（P〈0.05）.With IR group,CSA myocardial ischemia and reperfusion myocardial apoptosis rate,CaN activity and Fas/FasL protein levels were significantly decreased（P〈0.05）.Conclusion CSA preconditioning myocardial ischemia reperfusion injury in rats by inhibiting CaN signal reduces myocardial infarct size,and can effectively reduce the Fas/FasL protein expression,delaying myocardial ischemia-reperfusion injury,has a protective role in myocardial cells.