吗啡预处理对缺血再灌注损伤后大鼠心脏中Caspase-3表达及细胞凋亡的影响  

Morphine preconditioning reduces global ischemia/reperfusion injury via regulating Caspase-3 and cardiomyocyte apoptosis in isolated rat hearts

作　　者：朱瑞 金世云[1,2] 郭成晓[1,2] 张野[1,2] Zhu Rui;Jin Shiyun;Guo Chengxiao(Dept of Anesthesiology, The Second Affiliated Hospital of Anhui Medical University, Hefei 230601;Key Laboratory of Anesthesiology and Perioperative Medicine of Anhui Higher Education, Anhui Medical University, Hefei 230032)

机构地区：[1]安徽医科大学第二附属医院麻醉与围术期医学科,合肥230601 [2]安徽医科大学麻醉与围术期医学安徽普通高校重点实验室,合肥230032

出　　处：《安徽医科大学学报》2021年第7期1022-1026,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81801938、81970231);安徽医科大学校科研基金(编号:2017xkj047)。

摘　　要：目的探讨吗啡预处理(MP)对大鼠全心缺血再灌注损伤(IRI)后心肌组织中Caspase-3表达水平以及心肌细胞凋亡的影响。方法雄性成年SD大鼠51只随机分为3组:对照组(Sham组)、全心缺血再灌注组(IR组)、MP组(1μmol/L)。Sham组持续灌注195 min。IR组灌注K-H液45 min后制备离体全心缺血30 min再灌注10、60、120 min模型。MP组于缺血前灌注含吗啡K-H液和无吗啡K-H液各5 min,3个循环。氯化三苯基四氮唑(TTC)染色测定梗死区(IS)、缺血危险区(AAR)以及IS/AAR比值;化学比色法测定冠脉流出液乳酸脱氢酶(LDH)活性;Western blot及TUNEL染色分别测定心肌组织Caspase-3蛋白表达及细胞凋亡情况。结果与Sham组比较,IR组再灌注末心肌梗死体积百分比增加,再灌注后各时间点LDH活性、Caspase-3蛋白表达及心肌细胞凋亡比率均显著升高(P<0.05);与IR组比较,MP显著降低IS/AAR比值和再灌注后各时点LDH活性、Caspase-3蛋白表达以及心肌细胞凋亡比率(P<0.05);IR组与MP组均以再灌注10 min时LDH水平、再灌注60 min时Caspase-3蛋白表达和再灌注120 min时细胞凋亡比例最高。结论MP的心肌保护作用与降低再灌注前期Caspase-3蛋白表达及减轻再灌注后心肌细胞凋亡有关。Objective To investigate the effect of morphine pretreatment on the expression level of Caspase-3 and apoptosis of myocardial cells after ischemia reperfusion injury in rats.Methods A total of 51 male adult SD rats were randomly divided into 3 groups:the control group(Sham),ischemia-reperfusion group(IR)and morphine preconditioning group(1μmol/L,MP).The Sham group was perfused with K-H solution for 195 min.In IRgroup,K-H solution was infused for 45 min to prepare in vitro whole heart ischemia for 30 min,and 10,60 and 120 min post-ischemia reperfusion models were made.In MP group,K-H solution with morphine was perfused and K-H solution without morphine was perfused for 5 min each before ischemia,with a total of 3 cycles.Infarct volume(IS),area at risk(AAR)and IS/AAR were determined by 2,3,5-triphenyl-tetrazolium(TTC)staining.Lactic dehydrogenase(LDH)activity in coronary artery outflow was measured by chemical colorimetry.Caspase-3 and cardiomyocyte apoptosis were detected after 10,60 and 120 min of reperfusion via Western blot and TUNEL staining,respectively.Results Compared with sham group,the percentage of infarct volume at the end of reperfusion increased in IR group,LDH activity,Caspase-3 protein expression and cardiomyocyte apoptosis ratio significantly increased in IR group at each time point after reperfusion(P<0.05).Compared with IR group,morphine pretreatment significantly decreased IS/AAR ratio,LDH active Caspase-3 protein level and myocardial apoptosis ratio(P<0.05).LDH level at 10 min reperfusion,Caspase-3 protein expression at 60 min reperfusion and apoptosis rate at 120 min reperfusion were the highest in IR group and MP group.Conclusion The protective effect of morphine pretreatment on myocardium is associated with decreased expression of Caspase-3 protein at the early stage of reperfusion and decreased apoptosis of myocardial cells after reperfusion.

关 键 词：吗啡 CASPASE-3 细胞凋亡 缺血再灌注损伤 

分 类 号：R332[医药卫生—人体生理学] R971.2[医药卫生—基础医学]