CAJ | 학술논문

目的探讨丝氨酸-苏氨酸激酶(Akt)/内皮型一氧化氮合酶(eNOS)信号通路在鞘内远端吗啡预处理减轻在体大鼠心肌缺血-再灌注损伤和细胞凋亡中的作用.方法鞘内置管成功的雄性SD大鼠36只,随机(随机数字法)分为3组:假手术组(Sham组)、缺血-再灌注组(I/R组)和鞘内远端吗啡预处理组(RMPC组).采用缺血30 min再灌注120 min的方法制备心肌缺血-再灌注模型.RMPC组在缺血前30 min内经5 min鞘内输注吗啡l μg/kg(用生理盐水稀释到10 μL),停止5 min,共3个循环;I/R组给予等容量生理盐水.记录吗啡预处理前(基础值)、缺血前即刻、缺血30 min、再灌注120 min时MAP和HR以及MAP与HR的乘积(RPP).实验结束处死大鼠,取心肌组织,计算梗死区体积及梗死区面积与缺血危险区体积的比值(IS/AAR);采用TUNEL染色法检测心肌细胞凋亡,计算凋亡指数(AI);采用Western blot法测定心肌组织Akt、磷酸化Akt (p-Akt)和eNOS表达.结果与Sham组比较,I/R组的IS体积、IS/AAR、心肌细胞AI和心肌组织p-Akt表达均增加(P＜0.01),而心肌组织eNOS表达下调(P＜0.01);与I/R组比较,RMPC组IS体积、IS/AAR和心肌细胞AI均降低(P＜0.01),心肌组织p-Akt和eNOS表达上调(P＜0.01).结论鞘内远端吗啡预处理减轻在体大鼠心脏缺血后损伤和心肌细胞凋亡,其机制可能与Akt/eNOS信号通路参与介导有关.
목적 탐토사안산-소안산격매(Akt)/내피형일양화담합매(eNOS)신호통로재초내원단마배예처리감경재체대서심기결혈-재관주손상화세포조망중적작용.방법 초내치관성공적웅성SD대서36지,수궤(수궤수자법)분위3조:가수술조(Sham조)、결혈-재관주조(I/R조)화초내원단마배예처리조(RMPC조).채용결혈30 min재관주120 min적방법제비심기결혈-재관주모형.RMPC조재결혈전30 min내경5 min초내수주마배l μg/kg(용생리염수희석도10 μL),정지5 min,공3개순배;I/R조급여등용량생리염수.기록마배예처리전(기출치)、결혈전즉각、결혈30 min、재관주120 min시MAP화HR이급MAP여HR적승적(RPP).실험결속처사대서,취심기조직,계산경사구체적급경사구면적여결혈위험구체적적비치(IS/AAR);채용TUNEL염색법검측심기세포조망,계산조망지수(AI);채용Western blot법측정심기조직Akt、린산화Akt (p-Akt)화eNOS표체.결과 여Sham조비교,I/R조적IS체적、IS/AAR、심기세포AI화심기조직p-Akt표체균증가(P＜0.01),이심기조직eNOS표체하조(P＜0.01);여I/R조비교,RMPC조IS체적、IS/AAR화심기세포AI균강저(P＜0.01),심기조직p-Akt화eNOS표체상조(P＜0.01).결론 초내원단마배예처리감경재체대서심장결혈후손상화심기세포조망,기궤제가능여Akt/eNOS신호통로삼여개도유관.
Objective To investigate the effects of intrathecal morphine remote preconditioning (MRPC) on protein-serine-threonine kinases-endothelial nitric oxide synthase (Akt/eNOS) signaling pathways and cardiac myocyte apoptosis in rats.Methods Male SD rats weighing 280-320 g were used in this study.A needle was inserted through a surgically created hole into the sub-dural space of spinal cord.Thirty-six rats in which intrathecal needle was successfully placed without complication were randomly divided into 3 groups (n =12 in each).In group Ⅰ sham operation was performed (Sham).In group Ⅱ myocardial I/R was produced (I/R).In group Ⅲ morphine was given intrathecally in 3 repeated doses of 1 μg/kg at 5 min intervals before ischemia (MRPC).Myocardial I/R was produced by occlusion of left anterior descending branch (LAD) of coronary artery for 30 min followed by 120 min reperfusion.The animals were then sacrificed and hearts removed for measurement of area at risk (AAR) and infarct size area (IS).IS/AAR ratio was calculated.Myocardial apoptosis was detected by TUNEL and apoptotic index (the number of apoptotic myocardial cells/the total number of myocardial cells) was calculated.The levels of Akt,phosphorylated Akt (p-Akt) and eNOS was determined by Western blot.Results The infarct size,myocardial cell apoptotic index and pAkt level were higher and eNOS level was significantly lower in I/R group than those in group Sham (P ＜ 0.01).MRPC significantly reduced the infarct size and myocardial cell apoptotic index,and pAkt and eNOS level up-regulated in group RMPC compared with group I/R (P ＜ 0.01).Conclusions Akt/eNOS signaling pathways probably participate in the protective effects of intrathecal morphine remote preconditioning against myocardial I/R injury and myocardial cell apoptosis in rats.