CAJ | 학술논문

目的探讨单磷酸腺苷激活蛋白激酶(AMPK)信号通路在七氟醚预处理增强大鼠心肌细胞缺氧复氧时抗氧化能力中的作用.方法采用随机数字表法,将大鼠胚胎心肌细胞(H9C2细胞)分为3组(n=12):对照组(C组)、缺氧复氧损伤组(H/R组)和七氟醚预处理(SP组);采用随机数字表法,将AMPK基因敲除(AMPK siRNA)的H9C2细胞分为3组(n=12):对照组(A-C组)、缺氧复氧损伤组(A-H/R组)和七氟醚预处理(A-SP组).H9C2细胞置于95％N2+ 5％CO2培养箱孵育14 h,之后置于95％O2+ 5％CO2培养箱中孵育3h,建立缺氧复氧模型.SP组H9C2细胞培养液中加入七氟醚,终浓度为2.0 mmol/L,每次孵育15 min,共3次,间隔15 min进行七氟醚预处理.七氟醚预处理结束后15min建立缺氧复氧模型.复氧3h时,采用lucigenin增强化学发光法和DHE染色测定超氧阴离子水平,并测定LDH释放量和caspase-3活性.结果与C组比较,H/R组和SP组H9C2细胞LDH释放量增多,caspase-3活性增强,超氧阴离子水平升高(P＜0.01);与H/R组比较,SP组H9C2细胞LDH释放量减少,caspase-3活性减弱,超氧阴离子水平降低,A-H/R组H9C2细胞LDH释放量增多,caspase-3活性增强,超氧阴离子水平升高(P＜0.01);与SP组比较,A-SP组H9C2细胞LDH释放量增多,caspase-3活性增强,超氧阴离子水平升高(P＜0.01);与A-H/R组比较,A-SP组H9C2细胞LDH释放量减少,caspase-3活性减弱,超氧阴离子水平降低(P＜0.01).结论 AMPK信号通路参与了七氟醚预处理增强大鼠心肌细胞缺氧复氧时的抗氧化能力,但并未起主要作用.
목적 탐토단린산선감격활단백격매(AMPK)신호통로재칠불미예처리증강대서심기세포결양복양시항양화능력중적작용.방법 채용수궤수자표법,장대서배태심기세포(H9C2세포)분위3조(n=12):대조조(C조)、결양복양손상조(H/R조)화칠불미예처리(SP조);채용수궤수자표법,장AMPK기인고제(AMPK siRNA)적H9C2세포분위3조(n=12):대조조(A-C조)、결양복양손상조(A-H/R조)화칠불미예처리(A-SP조).H9C2세포치우95％N2+ 5％CO2배양상부육14 h,지후치우95％O2+ 5％CO2배양상중부육3h,건립결양복양모형.SP조H9C2세포배양액중가입칠불미,종농도위2.0 mmol/L,매차부육15 min,공3차,간격15 min진행칠불미예처리.칠불미예처리결속후15min건립결양복양모형.복양3h시,채용lucigenin증강화학발광법화DHE염색측정초양음리자수평,병측정LDH석방량화caspase-3활성.결과 여C조비교,H/R조화SP조H9C2세포LDH석방량증다,caspase-3활성증강,초양음리자수평승고(P＜0.01);여H/R조비교,SP조H9C2세포LDH석방량감소,caspase-3활성감약,초양음리자수평강저,A-H/R조H9C2세포LDH석방량증다,caspase-3활성증강,초양음리자수평승고(P＜0.01);여SP조비교,A-SP조H9C2세포LDH석방량증다,caspase-3활성증강,초양음리자수평승고(P＜0.01);여A-H/R조비교,A-SP조H9C2세포LDH석방량감소,caspase-3활성감약,초양음리자수평강저(P＜0.01).결론 AMPK신호통로삼여료칠불미예처리증강대서심기세포결양복양시적항양화능력,단병미기주요작용.
Objective To investigate the role of 5'-AMP-activated protein (AMPK) signaling pathway in sevoflurane preconditioning-induced enhancement of antioxidant capacity during hypoxia-reoxygenation (H/R) in the cardiomyocytes of the rats.Methods The rat embryonic cardiomyocytes (H9C2 cells) were randomly divided into 3 groups (n =12 each) using a random number table:control group (group C),H/R group,and sevoflurane preconditioning group (group SP).AMPK gene knockout (AMPK siRNA) H9C2 cells were randomly divided into 3 groups (n =12 each) using a random number table:control group (group A-C),A-H/R group,and sevoflurane preconditioning group (group A-SP).The cells were exposed to 95％ N2 + 5％ CO2 in an incubator for 14 h followed by reoxygenation with 95％ O2 + 5％ CO2 for 3 h.In SP group,the cells were preconditioned with 15 min sevoflurane (final concentration 2.0 mmol/L) three times at 15 min interval and H/R model was established at 15 min after the end of preconditioning.At 3 h of reoxygenation,the level of superoxide anion was detected using lucigenin-enhanced chemiluminescence and DHE staining,and the release of dehydrogenase (LDH) and caspase3 activity were measured.Results Compared with group C,the release of LDH and level of superoxide anion were significantly increased,and caspase-3 activity was enhanced in group H/R.Compared with group H/R,the release of LDH and level of superoxide anion were significantly decreased,and caspase-3 activity was weakened in group SP,and the release of LDH and level of superoxide anion were significantly increased,and caspase-3 activity was enhanced in group A-H/R.Compared with group SP,the release of LDH and level of superoxide anion were significantly increased,and caspase-3 activity was enhanced in group A-SP.Compared with group A-H/R,the release of LDH and level of superoxide anion were significantly decreased,and caspase-3 activity was weakened in group A-SP.Conclusion AMPK signaling pathway is involved in the mechanism by which sevoflurane preconditioning enhances the antioxidant capacity during H/R in the cardiomyocytes of the rats,but it does not play a major role.