牛磺酸抗心肌成纤维细胞增殖作用的实验研究

目的研究牛磺酸(Taurine,Tau)对血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)诱导新生大乳鼠心肌成纤维细胞(myofibroblasts,myo Fbs)增殖的抑制作用,并探讨其作用机制.方法用AngⅡ诱导新生大乳鼠myo Fbs增殖,建立心肌纤维化(myofibrosis,MF)模型.采用四甲基偶氮唑盐(MTT)法检测细胞增殖;羟脯氨酸试剂盒检测胶原含量;免疫细胞化学染色检测p-PKCα的膜转位及表达;Western blot检测p-PKCα和p-ERK1/2蛋白表达及含量.结果 Tau(30,60,120)mmol/L可明显抑制AngⅡ诱导的Myo Fbs增殖及胶原合成,同AngⅡ组比较差异具有统计学意义(P0.05或P0.01),并且呈现出剂量依赖性.应用免疫细胞染色Western blot并且结合图像分析,结果表明:Tau可抑制p-PKCα膜转位和表达,与AngⅡ组相比差异具有统计学意义(P0.01).结论 Tau可能通过减少p-PKCα的表达及膜转位,从而抑制MyoFbs增殖和胶原含量的增加,抑制MF,逆转心肌重塑.

Experimental Study on Effect of Taurine on the Anti-proliferation of Myofibroblasts

Objective To investigate the effect of Taurine( Tau) on inhibiting the proliferation of myoFbs of the neonatal rats induced by angiotensinⅡ( AngⅡ) , and to explore its mechanism.Methods The myoFbs proliferation of the cultured neonatal rat was induced by AngⅡ, and was detected with thiazole blue ( MTT ) colorimetric assay.The levels of collagenⅠand collagen Ⅲ were measured by hydroxyproline kit.Translocation and expression of p-PKCα in cells membrane or nuclear were determined by the immunohistochemistry staining with image analysis software.The expression and content of p-PKCαand p-ERK1/2 in cells were determined with western technology and gel imaging system.Results MyoFbs proliferation and hydroxyproline content in the culture medium were markedly inhibited when myoFbs were dealed with Tau at the concentrations of (120,60 and 30)mmol/L for 24 h(P<0.01,P<0.05,respectively),and showed dose-dependent.Immunocytochemistry, western blot and image analysis system showed that Tau could inhibit the expression and membrane translocation of p-PKCα, and further inhibit the nuclear translocation and expression of p-ERK1/2.These results showed statistically significant difference compared with AngⅡgroup(P<0.01).Conclusion These results suggest that Tau can inhibit the proliferation of myoFbs and the increase of collagen content by inhibiting the translocation and expression of p-PKCα in membrane,and then inhibit MF and reverse cardiac remodeling.