CAJ | 학술논문

目的探讨不同浓度七氟醚预处理对大鼠海马神经元缺氧复氧时细胞凋亡的影响及线粒体ATP敏感型钾通道(mito-KATP通道)在其中的作用.方法新生(出生＜24 h)SD大鼠,雌雄不拘,体重5～6 g,原代培养海马神经元,接种于培养孔或培养皿中,采用随机数字表法,将其随机分为7组,每组48孔和12皿,正常对照组(C组):不予任何处理;缺氧复氧组(HR组):缺氧4 h复氧24 h;6%七氟醚预处理组(S1 组)、4%七氟醚预处理组(S2 组)、2%七氟醚预处理组(S3 组):分别经6%、4%、2%七氟醚预处理后行缺氧复氧;5-羟葵酸100 μmol/L预处理组(5-HD组):经mito-KATP通道阻断剂5-羟葵酸(终浓度100 μmol/L)预处理后进行缺氧复氧;5-羟葵酸100 μmol/L+6%七氟醚预处理组(5-HD+S组):同时行5-羟葵酸和6%七氟醚预处理后进行缺氧复氧.各组以上处理结束后,测定神经元活力、凋亡率、Bcl-2和Bax蛋白的表达水平.结果与C组比较,其余6组海马神经元活力降低,细胞凋亡率升高,Bcl-2和Bax蛋白表达上调(P＜0.01);与HR组比较,S1组～S3组海马神经元活力增强,细胞凋亡率降低,Bcl-2蛋白表达上调,Bax蛋白表达下调(P＜0.01),5-HD组和5-HD+S组上述指标比较差异无统计学意义(P＞0.05);与S1组比较,S2组、S3组和5-HD+S组海马神经元活力降低,细胞凋亡率升高,Bcl-2蛋白表达下调,Bax蛋白表达上调(P＜0.01);与S2组比较,S3组海马神经元活力降低,细胞凋亡率升高,Bcl-2蛋白表达下调,Bax蛋白表达上调(P＜0.01).结论七氟醚预处理可抑制大鼠海马神经元缺氧复氧时细胞凋亡,从而减轻神经元损伤,且呈浓度依赖性,机制可能与开放神经元mito-KATP通道,上调Bcl-2蛋白表达,下调Bax蛋白表达有关.
목적 탐토불동농도칠불미예처리대대서해마신경원결양복양시세포조망적영향급선립체ATP민감형갑통도(mito-KATP통도)재기중적작용.방법 신생(출생＜24 h)SD대서,자웅불구,체중5～6 g,원대배양해마신경원,접충우배양공혹배양명중,채용수궤수자표법,장기수궤분위7조,매조48공화12명,정상대조조(C조):불여임하처리;결양복양조(HR조):결양4 h복양24 h;6%칠불미예처리조(S1 조)、4%칠불미예처리조(S2 조)、2%칠불미예처리조(S3 조):분별경6%、4%、2%칠불미예처리후행결양복양;5-간규산100 μmol/L예처리조(5-HD조):경mito-KATP통도조단제5-간규산(종농도100 μmol/L)예처리후진행결양복양;5-간규산100 μmol/L+6%칠불미예처리조(5-HD+S조):동시행5-간규산화6%칠불미예처리후진행결양복양.각조이상처리결속후,측정신경원활력、조망솔、Bcl-2화Bax단백적표체수평.결과 여C조비교,기여6조해마신경원활력강저,세포조망솔승고,Bcl-2화Bax단백표체상조(P＜0.01);여HR조비교,S1조～S3조해마신경원활력증강,세포조망솔강저,Bcl-2단백표체상조,Bax단백표체하조(P＜0.01),5-HD조화5-HD+S조상술지표비교차이무통계학의의(P＞0.05);여S1조비교,S2조、S3조화5-HD+S조해마신경원활력강저,세포조망솔승고,Bcl-2단백표체하조,Bax단백표체상조(P＜0.01);여S2조비교,S3조해마신경원활력강저,세포조망솔승고,Bcl-2단백표체하조,Bax단백표체상조(P＜0.01).결론 칠불미예처리가억제대서해마신경원결양복양시세포조망,종이감경신경원손상,차정농도의뢰성,궤제가능여개방신경원mito-KATP통도,상조Bcl-2단백표체,하조Bax단백표체유관.
Objective To investigate the effect of preconditioning with different concentrations of sevoflurane on hypoxia-reoxygenation(H/R)-induced apoptosis in rat hippocampal neurons and the role of mitochondrial KATP(mito-KATP)channels.Methods Primary cultured hippocampal neurons isolated from newborn SD rats(＜24h)of both sexes,weighing 5-6 g,were randomly divided into 7 groups with 48 wells and 12 dishes in each one:control group(C group),H/R group,preconditioning with 6%,4%and 2% sevoflurane groups(S1-3 groups),5-hydroxydecanoate(5-HD,mito-KATP channel blocker)100 μmol/L preconditioning group(5-HD group)and preconditioning with 5-HD 100 μmol/L+6% sevoflurane group(5-HD+S group).The neurons were exposed to 4 h hypoxia followed by 24 h reoxygenation. In S1-3 groups, preconditioning was performed with 6% , 4% and 2% sevoflurane respectively before H/R. In 5-HD group, preconditioning was performed with 5-HD (final concentration 100 μmol/L) before H/R. In 5-HD + S group, preconditioning was performed with 5-HD 100 μmol/L and 6% sevoflurane before H/R. The neuronal viability, apoptosis rate and expression of Bcl-2 and Bax were determined after 24 h reoxygenation.Results The neuronal viability was significantly lower,while the apoptosis rate and expression of Bcl-2 and Bax were significantly higher in the other 6 groups than in group C(P＜0.01).The neuronal viability and expression of Bcl-2 were significantly higher,while the apoptosis rate and Bax expression were lower in S1-3 groups than in group H/R. There was no significant difference in the parameters mentioned above between 5-HD and 5-HD + S groups(P＞0.05).The neuronal viability and expression of Bcl-2 were significantly lower, while the apoptosis rate and Bax expression were higher in S2, S3 and 5-HD + S groups than in group S1, and in group S3 than in group S2(P＜0.0l) .Conclusion Sevoflurane preconditioning can inhibit H/R-induced apoptosis in rat hippocampal neurons and reduce the injury to neurons in a concentration-dependent manner, and the underlying mechanism may be related to activation of mito-KATP channels, up-regulation of Bcl-2 expression and down-regulation of Bax expression.