牛磺酸对大乳鼠心肌细胞抗氧化作用的荧光免疫化学分析

Fluorescence Immunoassay and Chemical Analysis of Effect of Taurine on Neonatal Rat Myocardial Cells Antioxidant Role

应用免疫和化学分析方法探讨牛磺酸(Taurine,Tau)对原代培养的新生大乳鼠心肌细胞的保护作用,为心血管疾病的治疗提供理论依据。将原代培养的乳鼠心肌细胞随机分为对照组、过氧化氢(H2O2)损伤模型组以及H2O2损伤加药物治疗组。预先12 h给予Tau治疗药物后,使用H2O2损伤心肌细胞4 h,在倒置显微镜观察Tau对心肌细胞形态学的影响,3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(3-(4,5)-Dimethyl-thiahiazo(-z-yl)-3,5-diphenytetrazoliumromide,MTT)化学分析法测定心肌细胞的存活率,化学试剂盒检测心肌细胞内超氧化物歧化酶(Superoxide dismutase,SOD)活性及丙二醛(Malondialdehyde,MDA)含量的变化,利用荧光探针DCFH-DA分析检测心肌细胞中活性氧的水平。结果表明,H2O2损伤心肌细胞4 h后,Tau可使吸光度升高,并增加心肌细胞SOD活性、降低MDA含量及活性氧水平下降。由此可见,采用荧光免疫分析方法可证实Tau能减轻自由基对心肌细胞的损伤,从而达到保护心脏的作用。

The protective effect of taurine（Tau） on primary cultured neonatal rat myocardial cells was investigated by immune and chemical analysis methods to obtain the theoretical basis for the treatment of cardiovascular disease.The myocardial cells were randomly divided into below groups： control group,H2O2 damage group and taurine groups.The morpholoical changes of myocardial cells damaged for 4 h by 300 μm/L H2O2 under the inverted microscope were observed.Meanwhile,myocardial cell survival rate was measured by 3-（4,5）-dimethyl-thiahiazo（-2-yl）-3,5-diphenyl-tetrazoliumromide（MTT） chemical analysis method,superoxide dismutase（SOD） activity and malondialdehyde（MDA） content changes were detected by chemical reagent kit,the level of reactive oxygen species（ROS） in myocardial cell were measured by the way of fluorescence probe 2′7′-dichlorofluorescin diacetate（DCFH-DA） assays.The results showed that,after myocardial cells were injured for 4 h by H2O2,Tau could increase the A value and SOD activity in myocardial cells,and reduce the content of MDA and decline reactive oxygen species levels.The fluorescence immunoassay method can prove that Tau could reduce free radical on myocardial cells injury to cardiac protective effect.