CAJ | 학술논문

目的评价线粒体ATP敏感性钾通道(mito-KATP通道)在七氟醚预处理对大鼠离体缺血再灌注心脏延迟性保护中的作用.方法健康成年雄性SD大鼠72只,体重270～350 g,采用随机数字表法,将其随机分为6组(n=12):对照组(CON组)、缺血再灌注组(I/R组)各吸入33％氧气1h；七氟醚对照组(SEVO组)、七氟醚预处理组(SWOP组)各吸入2.5％七氟醚1h;5-羟葵酸+七氟醚预处理组(5-HD+ SWOP组)于七氟醚预处理前30 min腹腔注射mito-KATP阻断剂5-羟葵酸10 mg/kg;5-羟葵酸对照组(5-HD组)腹腔注射5-羟葵酸10 mg/kg,24 h后除CON组和SEVO组,其余组建立大鼠Langendorff离体心脏灌注模型,停止灌注30 min,再灌注120 min.于缺血前即刻(T0)和再灌注120 min(T1)时采用Western blot法检测心肌组织磷酸化的ε型蛋白激酶C(p-PKC-ε)、caspase-8蛋白的表达水平；于再灌注120 min时采用TTC法测定心肌梗死范围,计算心肌梗死体积百分比.结果与CON组比较,I/R组、SWOP组、5-HD+ SWOP组和5-HD组T1时心肌梗死体积百分比升高,SEVO组和SWOP组T0时,I/R组、SWOP组、5-HD+ SWOP组和5-HD组T1时心肌p-PKC-ε蛋白表达上调,SEVO组T2时caspase-8 蛋白表达下调(P＜0.05)；与I/R组比较,SWOP组和5-HD+ SWOP组T1时心肌梗死体积百分比降低,SEVO组和SWOP组T0时p-PKC-ε蛋白表达上调,SWOP组T1时caspase-8蛋白表达下调(P＜ 0.05)；与SWOP组比较,5-HD+ SWOP组心肌梗死体积百分比升高,T0时p-PKC-ε蛋白表达下调,T1时caspase-8 蛋白表达上调(P＜0.05).结论 mito-KATP通道参与了七氟醚预处理减轻大鼠离体心脏缺血再灌注损伤的过程,可能与上调缺血前p-PKC-ε蛋白表达水平,从而抑制再灌注时细胞凋亡有关.
목적 평개선립체ATP민감성갑통도(mito-KATP통도)재칠불미예처리대대서리체결혈재관주심장연지성보호중적작용.방법 건강성년웅성SD대서72지,체중270～350 g,채용수궤수자표법,장기수궤분위6조(n=12):대조조(CON조)、결혈재관주조(I/R조)각흡입33％양기1h；칠불미대조조(SEVO조)、칠불미예처리조(SWOP조)각흡입2.5％칠불미1h;5-간규산+칠불미예처리조(5-HD+ SWOP조)우칠불미예처리전30 min복강주사mito-KATP조단제5-간규산10 mg/kg;5-간규산대조조(5-HD조)복강주사5-간규산10 mg/kg,24 h후제CON조화SEVO조,기여조건립대서Langendorff리체심장관주모형,정지관주30 min,재관주120 min.우결혈전즉각(T0)화재관주120 min(T1)시채용Western blot법검측심기조직린산화적ε형단백격매C(p-PKC-ε)、caspase-8단백적표체수평；우재관주120 min시채용TTC법측정심기경사범위,계산심기경사체적백분비.결과 여CON조비교,I/R조、SWOP조、5-HD+ SWOP조화5-HD조T1시심기경사체적백분비승고,SEVO조화SWOP조T0시,I/R조、SWOP조、5-HD+ SWOP조화5-HD조T1시심기p-PKC-ε단백표체상조,SEVO조T2시caspase-8 단백표체하조(P＜0.05)；여I/R조비교,SWOP조화5-HD+ SWOP조T1시심기경사체적백분비강저,SEVO조화SWOP조T0시p-PKC-ε단백표체상조,SWOP조T1시caspase-8단백표체하조(P＜ 0.05)；여SWOP조비교,5-HD+ SWOP조심기경사체적백분비승고,T0시p-PKC-ε단백표체하조,T1시caspase-8 단백표체상조(P＜0.05).결론 mito-KATP통도삼여료칠불미예처리감경대서리체심장결혈재관주손상적과정,가능여상조결혈전p-PKC-ε단백표체수평,종이억제재관주시세포조망유관.
Objective To evaluate the role of the mitochondrial ATP-sensitive potassium (mito-KATP)channel in sevoflurane preconditioning-induced delayed cardioprotection against ischemia-reperfusion (I/R) injury in isolated rat hearts.Methods Seventy-two adult male Sprague-Dawley rats were randomly divided into 6 groups (n =12 each):control group (group CON),I/R group,sevoflurane control group (group SEVO),sevoflurane preconditioning group (group SWO P),5-hydroxydeconoate (5-HD) + sevoflurane preconditioning group (group 5-HD+ SWOP) and 5-HD control group (group 5-HD).The rats were exposed to 33％ pure oxygen for 1 h in groups CON and I/R.The rats were exposed to 2.5％ sevoflurane for 1 h in groups SEVO and SWOP.5-HD (a mito-KATP channel inhibitor) 10 mg/kg was injected intraperitoneally 30 min before sevoflurane preconditioning in group 5-HD + SWOP.5-HD 10 mg/kg was injected intraperitoneally in group 5-HD.The hearts were immediately removed and perfused in a Langendorff apparatus.The hearts were made globally ischemic for 30 min followed by 120 min reperfusion in groups I/R,SWOP,5-HD + SWOP and 5-HD.The expression of phosphorylated protein kinase C-epsilon (p-PKC-ε) and caspase-8 was measured by Western blot immediately before ischemia (T0) and at 120 min of reperfusion (T1).The myocardial infarct volume was measured by TTC staining.Results Compared with group CON,the myocardial infarct volume was significantly increased at T1 in groups I/R,SWOP,5-HD +SWOP and 5-HD,p-PKC-ε expression was up-regulated at T0 in groups SEVO and SWOP and at T1 in groups I/R,SWOP,5-HD + SWOP and 5-HD,and caspase-8 expression was down-regulated at T1 in group SEVO (P ＜0.05).Compared with group I/R,the myocardial infarct volume was significantly decreased at T1 in groups SWOP and 5-HD + SWOP,p-PKC-ε expression was up-regulated at T0 in groups SEVO and SWOP,and caspase-8 expression was down-regulated at T1 in group SWOP (P ＜ 0.05).Compared with group SWOP,the myocardial infarct volume was significantly increased,p-PKC-ε expression was down-regulated at T0,and caspase-8 expression was up-regulated at T1 in group 5-HD + SWOP (P ＜ 0.05).Conclusion The mito-KATP channel is involved in sevoflurane preconditioning-induced delayed cardioprotection against I/R injury in isolated rat hearts through upregulation of p-PKC-ε expression before ischemia and inhibition of cell apoptosis during reperfusion.