MitoKATP与枳实薤白桂枝汤保护家兔心肌缺血再灌注损伤机制相关性研究

摘要

目的 目的通过实验研究观察线粒体ATP敏感性钾离子通道(MitoKATP)是否参与枳实薤白桂枝汤(ZSXBGZD)对家兔心肌缺血再灌注损伤(MIRI)保护性机制.方法 将40只大耳家兔随机分为5组,每组8只,即非缺血再灌注组(Sham组)、缺血再灌注组(MIRI组)、缺血再灌注+枳实薤白桂枝汤组(ZSXBGZD组)、缺血再灌注+枳实薤白桂枝汤+5-羟基癸酸盐组(5-HD组)、缺血再灌注+枳实薤白桂枝汤+格列本脲组(Gli组),每组各8只.利用Langendorff体外法备MIRI模型,Sham组家兔完成全部操作,不进行停灌,持续灌流200 min;其余4组家兔离体心脏平衡灌流30 min后,进行停灌80 min;ZSXBGZD组于缺血前10 min给予含有枳实薤白桂枝汤提取液的locke液灌流10 min,然后缺血80 min,再灌注时仍给予含有枳实薤白桂枝汤提取液的locke液灌流120 min;5-HD组、Gli组过程同ZSXGZD组,在给予ZSXBGZD药物的基础上分别再加入5-HD、Gli.灌流结束后测定心肌组织乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)含量,分别记录停灌前10 min及再灌注后20、40、60、80、100、120 min各组家兔的冠脉流量,利用2,3,5-氯化三苯基四氮唑法(TTC法)测定心肌梗死面积.结果 MIRI组中家兔MDA、LDH、心肌梗死面积均明显高于Sham组,SOD、冠脉流量明显低于Sham组,比较差异均有统计学意义(P<0.05);与MIRI组相比,ZSXBGZD组SOD、冠脉流量均明显增加,心肌梗死面积缩小,MDA、LDH降低,比较差异均有统计学意义(P<0.05).与MIRI组比较,5-HD组及Gli组SOD、MDA、LDH含量,冠脉流量及心肌梗死面积均无明显变化(P>0.05),但与ZSXBGZD组比较差异均有统计学意义(P<0.05).结论 ZSXBGZD能减轻家兔MIRI损伤,具有保护心肌作用,且此作用能被5-HD大部分抵消,能被Gli完全阻断,提示枳实薤白桂枝汤对家兔MIRI保护作用与MitoKATP显著性相关.%Objective To investigate the correlation between MitoKATP and the mechanism of Zhishi-xie-bai-guizhi decoction (ZSXBGZD)in protection of myocardial ischemia reperfusion injury (MIRI)rabbits. Meth-ods 40 rabbits were randomly divided into 5 groups,including the non-ischemic reperfusion group (Sham group),ischemia-reperfusion group (MIRI group),ischemia and reperfusion with ZSXBGZD group (MIRI + ZSX-BGZD group),ischemia and reperfusion with ZSXBGZD + 5-HD group (5-HD group),ischemia and reperfusion ZSXBGZD + Gli group (Gli group),8 rabbits in each group. MIRI model was established by Langendorff isolated heart perfusion system. Rabbits in Sham group received o-verall operation,continuous perfusion 200 min without ische-mia. Rabbits other four groups,after 30 min stabilization the injury was induced by 80 min ischemia. ZSXBGZD group re-ceived 10 min perfusion of locke's liquid with ZSXBGZD ex-tracting solution 10 min before ischemia,ischemia for 80 min and 120 min reperfusion of locke's liquid with ZSXBGZD ex-tracting solution. The 5-HD and Gli was added in 5-HD and Gli group on the basis of ZSXGAD group operation. After perfusion,the lactic dehydrogenase (LDH),malonaldehyde (MDA)and superoxide dismutase (SOD)in car-diac muscle tissue were measure. The coronary flow rate was recorded at 10 min before ischemia,and 20 min,40 min,60 min,80 min,100 min,120 min after reperfusion. The myocardial infarction area was measured by 2,3,5 -triphenyltetrazolium chloride(TTC)method. Results The MDA,LDH and myocardial infarction area in MIRI group were obvious higher than Sham group,the SOD and coronary flow rate were obvious lower,with statistical differences (P < 0. 05). As compared with MIRI,the SOD and coronary flow rate in ZSXBGZD group were markedly increased,and the myocardial infarction area was decreased,the MAD and LDH were decreased,with statistical differences (P < 0. 05). As compared with MIRI group,there were no statistical differences on the SOD,MDA, LDH,coronary flow rate and myocardial infarction area in 5-HD and Gli group (P > 0. 05),but there were statisti-cal differences with ZSXBGZD group (P < 0. 05). Conclusion ZSXBGZD can reduce injury of MIRI in rabbits, has protect effect of myocardium,which can be largely offset by 5-HD and can be completely blocked by Gli, showed that the protective effects of Zhishi-xiebai-guizhi decoction on MIRI rabbits were significantly associated with MitoKATP.