白介素10对肿瘤坏死因子-α诱导心肌细胞凋亡的影响及其作用机制  

The Role of Apoptosis-Inducing Factor on TNF-α Induced Apoptosis in Cultured Neonatal Rat Cardiontyocytes

作　　者：张闻多[1] 马青变[2] 史力斌[3] 丁文惠[3] 

机构地区：[1]北京医院心内科,北京市100030 [2]北京大学第三医院急诊科 [3]北京大学第一医院心内科

出　　处：《中国基层医药》2010年第11期1444-1446,共3页Chinese Journal of Primary Medicine and Pharmacy

摘　　要：目的 探讨凋亡诱导因子（AIF）在肿瘤坏死因子-α（TNF-α）诱导心肌细胞凋亡途径中的作用,以及白介素-10（IL-10）对TNF-α诱导心肌细胞凋亡的影响及其可能的作用机制.方法 原代培养的乳鼠心肌细胞,分为六组（对照组、TNF-α 100 ng/ml 12 h、18 h和24 h组,IL-10 50ng/ml 19 h、IL-10 50.g/ml＋TNF-α 100 ng/ml 18 h组）.用流式细胞仪和Hoeehest 33258染色检测心肌细胞凋亡;weatertrl blot检测AIF蛋白的表达.结果 TNF-α 100ng/ml 12 h、18 h、24 h组心肌细胞的凋亡率较对照组均明显升高：12 h组流式细胞仪法（5.08±0.79）%与（2.2±0.77）%;18 h（14.39±2.31）%与（2.2±0.77）%;24（4.61±0.84）%与（2.2±0.77）%.18 h组Hoechst 33258法（18.936±2.79t）%与（2.890±1.326）%,P〈0.01;18 h凋亡率达高峰,明显高于12 h、24 h组[分别为（14.39±2.31）%与（5.08±0.79）%;（14.39±2.31）%与（4.61±0.84）%]加入TNF-α后12 h、18 h、24 h,较对照组AIF蛋白的表达明显增加,18h的AIF蛋白的表达增加更明显.结论AIF可能是TNF-α诱导心肌细胞凋亡的途径之一;IL-10对TNF-α诱导的心肌细胞凋亡有保护作用.Objective This study was designed to investigate the role of apoptosis-inducing factor on neonatal rat cardiomyocytes apoptosis induced by TNF-a,and to detect the effect of IL-10 on the apoptosis rate of cardio-myocytes apoptosis induced by TNF-α.Methods Neonatal rat cardiacmyocytes in primary culture were exposed to TNF-α（ 100ng/ml）for 12 hours,18 hours,24 hours,TNF-α（ 100ng/ml） for lohours pretreated by IL-10（50ng/ml） 1 hour,or IL-10（50ng/ml） for 19hours.Apoptotic cells were detected by flow cytometry and Hoechest 33258 dye; Western blot were used to determine the apoptosis-inducing factor protein expression.Results The apoptosis rate of cardiomyocytes increased in TNF-α 12 hours group than that of control[ Flow cytometry （5.08 ±0.79% ） vs （ 2.2± 0.77）% ,P 〈0.05],and reach the peak in TNF-α 18 hours group compared with control[ Flow cytometry（14.39± 2.31）%vs（2.2±0.77）%,P〈0.01;Hoechst 33258 dyeing（18.936±2.791）% Vs（2.890± 1.326）% ,P〈 0.01];also elevated in TNF-α 24 hours group vs control[Flow cytometry（4.61±0.84）% vs（2.2 ±0.77）% ,P〈 0.05].AIF expression：Increased expression of AIF in the cells treated by TNF-α（ 100 ng/ml） 12 hours,reached highest in TNF-α（ 100 ng/ml） for 18 hours group and decreased in TNF-α（ 100 ng/ml） 24 hours group compared with control;AIF expression is not decrease in TNF-α（ 100 ng/ml） 18 hours pretreated by IL-10（50 ng/ml） for 1-hour group comparing with TNF-α（ 100 ng/ml） 18hours group.Conclusion The study demonstrated that AIF contributed to TNF-α induced apoptosis of rat cardiomyocytes; IL-10 alleviated the apoptosis of rat cardiacmyocytes induced by TNF-α,and AIF may not its mechanism.

关 键 词：小鼠 心肌 白细胞介素10 肿瘤坏死因子 细胞凋亡 

分 类 号：R54[医药卫生—心血管疾病]