睡眠剥夺对缺血再灌注后心肌Caveolin-3表达与细胞凋亡的影响

Effect of sleep deprivation on caveolin-3 expression and apoptosis in the themyocardium after ischemia/reperfusion

目的:探究睡眠剥夺对缺血再灌注(I/R)后心肌Caveolin-3(Cav-3)表达与细胞凋亡的影响。方法:采用改良多平台水环境法建立小鼠急性睡眠剥夺模型,睡眠剥夺4 d后通过结扎小鼠冠状动脉左前降支45 min,随后再灌注120 min建立心肌I/R模型。采用苏木精-伊红染色法观察小鼠心肌病理改变,TTC染色测定I/R后小鼠心肌梗死面积,ELASA法测量血清肌酸激酶同工酶(CK-MB)含量,脱氧核糖核苷酸转移酶介导的dUTP缺口末端标记法(TUNEL法)检测细胞凋亡,Western Blot法检测Cleaved caspase-3和Cav-3的表达水平。结果:与对照组比较,睡眠剥夺4 d后小鼠变得易怒、激惹,病理学观察提示睡眠剥夺后心肌组织纤维排列紊乱,进一步研究发现小鼠睡眠剥夺后血浆CK-MB含量及心肌细胞凋亡水平明显增加(TUNEL染色阳性细胞与Cleaved caspase-3水平均显著增加),而心肌Cav-3表达水平明显降低。在睡眠剥夺小鼠进一步遭受心肌I/R后,其心肌梗死面积、CK-MB含量及细胞凋亡水平进一步增加,而Cav-3表达进一步下调(P<0.05)。结论:睡眠剥夺可导致心肌更不易耐受缺血再灌注损伤,其机制可能与Cav-3表达下调及心肌细胞凋亡过度激活有关。

Objective:To investigate the effects of sleep deprivation on the expression of myocardial caveo-lin-3(Cav-3)and apoptosis after ischemia/reperfusion(I/R).Methods:The acute sleep deprivation model of mouse was established by using the improved multi platform water environment method.Af-ter four days of sleep deprivation,the myocardial I/R model was established by ligating the left anteri-or descending coronary artery of mice for 45 minutes,and then reperfusion for 120 minutes.Hematox-ylin eosin staining was used to observe the pathological changes of mouse myocardium,TTC staining was used to determine the myocardial infarction area of mice after I/R,ELASA method was used to measure the content of serum creatine kinase isoenzyme(CK-MB),deoxyribonucleotidyltransferase mediated dUTP nick end labeling(TUNEL)method was used to detect apoptosis,and Western Blot method was used to detect the expression level of Cleaved caspase-3 and Cav-3.Results:Compared with the mice in control group,the mice became irritable and irritated after four days of sleep depriva-tion.Pathological observation showed that the arrangement of myocardial tissue fibers was disordered after sleep deprivation.Further research found that the content of CK-MB in plasma and the level of apoptosis of myocardial cells in mice after sleep deprivation were significantly increased(TUNEL staining positive cells and the level of Cleaved caspase-3 were significantly increased),while the level of expression of Cav-3 in myocardium was significantly reduced.After sleep deprived mice suffered further myocardial I/R,and their myocardial infarction area,CK-MB content and apoptosis level in-creased,while the expression of Cav-3 decreased(P<0.05).Conclusion:Sleep deprivation can make myocardium more resistant to ischemia reperfusion injury,and its mechanism may be related to the down-regulation of Cav-3 expression and excessive activation of myocardial cell apoptosis.