芬太尼和脂多糖迟发预处理对缺血再灌注损伤大鼠心肌细胞凋亡的影响

目的诱导大鼠心肌迟发预处理保护作用,测定心肌细胞凋亡率、Caspase、细胞色素C的变化,探讨DPC对心肌细胞凋亡的影响及作用机制。方法健康SD大鼠32只,随机分为4组,每组8只。芬太尼组（A组）：大鼠腹腔注射芬太尼,24小时后建立缺血再灌注损伤模型;脂多糖组（B组）：腹腔注射脂多糖,24小时后处理同A组。生理盐水组（C组）：腹腔注射生理盐水,24小时后处理同A组。对照组（D组）：不注射任何药物,24小时后处理同A组。TUNEL法检测心肌细胞凋亡;caspase-3试剂盒测定其活性;Westernblot法检测细胞色素C。结果HE染色：损伤重区域的心肌仍保存细胞轮廓,横纹不清或消失,收缩带形成。损伤轻区域细胞轮廓清晰,横纹不消失。TUNEL染色：散在的细胞核呈棕色凋亡细胞。A组和B组心肌细胞凋亡率分别为（9±3）%和（11±3）%低于C组（17±5）%;A组和B组caspase-3活性（分别为0.43±0.11和0.40±0.13）低于C组（0.75±0.23）;A组和B组细胞色素C蛋白表达（分别为0.67±0.11和0.63±0.18）低于C组（0.98±0.19）,均有显著差异（P〈0.05）。结论芬太尼、脂多糖诱导的迟发预处理中存在心肌细胞凋亡,且心肌细胞凋亡率、细胞色素C、Caspase-3降低。

Role of delayed preconditioning on apoptosis of rat myocardium induced by fentanyl and lipopolysaccharide

Objective To measure the apoptosis rate,Actavity of caspase-3,and Cytochrome C in the process of delayed preconditioning on rat myocardium,try to reveal their association and to manifest the role of delayed preconditioning on apoptosis of rat myocardium.Methods Thirty two SD rats were randomized into four groups（n=8）,fentanyl group（group A） were admin-istrated with fentanyl by peritoneal injection,then the heart underwent ischemic reperfusion injury in vivo after 24 hours.LPS group（group B）and NS group（group C）were treated with LPS and NS respectively,control group with nothing,they were under-went the same thing just like group A.Measure the apoptosis rate with TUNEL,actavity of caspase-3 and cytochrome C by west-ern blot.Results HE staining displays severe injuryed zone has normal outline without transverse striation.In mild injuryed zone there has obvious outline with transverse striation.TUNEL staining displayed scattered apoptotic body.the myocardium apoptosis rate,actavity of caspase-3 and cytochrome C expression in group A（9±3）%,0.43±0.11,0.67±0.11 and group B（11± 3）%,0.40±0.13,0.63±0.18 were significantly lower compared with that of group C respectively,（P〈0.05）.Conclusion There is myocardium apoptosis DPC induced by fentanyl or LPS,and apoptosis,Cytochrome C,caspase-3 decrease in this process.