同型半胱氨酸激活JNK信号通路诱导血管平滑肌细胞氧化应激的损伤研究

Oxidative stress induced by homocysteine viaactivating JNK signaling pathway in vascular smooth muscle cells

目的观察同型半胱氨酸(Hcy)在A10血管平滑肌细胞株(vascular smooth muscle cells,VSMC)上对氧化应激的影响及其可能的分子机制。方法在体外培养的A10细胞上,用3个不同浓度(5、30和100μmol/L)的Hcy孵育细胞共48h,随后破裂细胞,一方面使用试剂盒检测细胞裂解液中的活性氧(ROS)水平、H_2O_2含量、总抗氧化能力(T-AOC),以评价其氧化应激情况。另一方面,用免疫印迹法检测磷酸化应激活化蛋白激酶(JNK)及总JNK的表达量。结果Hcy可诱导ROS、H_2O_2的含量升高,而导致T-AOC的水平下降。而且,Hcy增加了磷酸化JNK的蛋白表达量(P<0.05)。结论 Hcy在VSMC上通过激活JNK信号通路诱导氧化应激。

Objective To investigate the influence and the underlying mechanisms of homocysteine on oxidative stress production in vascular smooth muscle cells（VSMC）.Methods A10 cell line（a kind of VSMC）was cultured in vitro.Three different concentrations of homocysteine（5,30 and 100μmol/L）were added into the medium for 48 hours and then cells were lysed for determination of reactive oxygen species（ROS）production,H2 O2 level and total anti-oxidant capacity.Moreover,the effects of homocysteine on JNK signaling pathway were evaluated by immunoblotting.Results ROS production（P〈0.05）in VSMC were induced by three different concentrations of homocysteine（5,30 and 100μmol/L）.Two concentrations of homocysteine（30 and 100μmol/L）increased H2 O2 content and decreased T-AOC.Moreover,homocysteine induced phosphorylation of JNK in VSMC in a dose-dependent manner（P〈0.05）.Conclusion Oxidative stress induced by homocysteine viaactivating JNK signaling pathway in VSMC.