摘要
为研究与分析牛磺酸干预对高糖培养H9c2心肌细胞的保护效应及其具体机制。通过将复苏的H9c2心肌细胞传至3～4代,分为以下三组:(1)对照组(CN):葡萄糖浓度为5. 5 mmol/L;(2)高糖组(HG):葡萄糖浓度为25. 5 mmol/L;(3)牛磺酸干预组(TAU):HG组基础上,加终浓度为40 mmol/L的牛磺酸。48 h后收集细胞,采用Annexin-V/碘化丙啶(propidine iodide,PI)结合流式细胞仪检测心肌细胞凋亡发生情况,Western Blot检测丝裂原活化蛋白激酶p38 MAPK的蛋白表达,放射免疫法检测TNF-α表达量,并采用相关试剂盒检测丙二醛(MDA)、超氧化物歧化酶(SOD)水平。结果显示:与CN组相比较,HG组H9c2心肌细胞经高糖培养48 h后细胞凋亡率增加(P <0. 001),细胞TNF-α、MDA显著水平升高(P <0. 001),SOD水平降低(P <0. 001),胞内磷酸化p38水平亦升高(P <0. 001);加入牛磺酸干预后,与HG组比较,TAU组细胞凋亡率减小(P <0. 01),细胞TNF-α、MDA及磷酸化p38蛋白水平明显降低(P <0. 05),SOD水平升高(P <0. 01)。可见牛磺酸能够减轻高糖刺激下H9c2心肌细胞的凋亡,其机制可能通过上调SOD水平,下调TNF-α、MDA水平,降低胞内p38磷酸化来实现对高糖细胞损伤的保护效应。
In order to investigate the protective effects and mechanisms of taurine on injured H9c2 myocardial cellsby high glucose.The resurrected H9c2 myocardial cells transmitted to 3~4 generations,were divided into three groups.①Control group(CN):glucose concentration was 5.5 mmol/L.②High glucose group(HG):glucose concentration was 25.5 mmol/L.③Taurine intervention group(TAU):on the basis of HG group,the final concentration of taurine was 40 mmol/L.After 48 h,the cells were collected and the apoptosis of myocardial cells was detected by Annexin-V/propidine iodide(PI)combined flow cytometry.The protein expression of mitogen activated protein kinase p38 MAPK was detected by Western Blot.The expression of TNF-αwas detected by radioimmunoassay,and the level of malonaldehyde(MDA)and superoxide dismutase(SOD)was detected by ELISA methods.The results showed that compared with the CN group,in HG group the apoptosis rate of H9c2 myocardial cellsincreased(P<0.001),the levels of TNF-αand MDA were also increased(P<0.001),the activity of SOD was decreased significantly(P<0.001)and the level of phosphorylation p38 was increased(P<0.001);compared with the HG group,in TAU group the apoptosis rate of H9c2 myocardial cells decreased(P<0.01),the level of TNF-α,MDA and phosphorylated p38 protein decreased(P<0.05),and the activity of SOD was increased(P<0.01).It is concluded that taurine can reduce the apoptosis of H9c2 cardiomyocytes stimulated by high glucose,and the mechanism may be related to the protection of high glucose cell injury by up-regulated level of SOD,down-regulated of TNF-αand MDA and the reduced intracellular phosphorylated p38.
作者
朱宁
张征
李良同
李少春
刘莉
周玉娟
张喆
ZHU Ning;ZHANG Zheng;LI Liang-tong;LI Shao-chun;LIU Li;ZHOU Yu-juan;ZHANG Zhe(Baoding First Center Hospital,Baoding 071000,China;Medical College,Baoding 071000,China;Hebei University,Baoding 071000,China)
出处
《科学技术与工程》
北大核心
2018年第32期157-161,共5页
Science Technology and Engineering
基金
河北省卫生和计划生育委员会青年科技课题(20120136)资助
