罗格列酮对HUVECs受损的保护作用及对TNF-a和IL-6水平的影响

Protective effect of rosiglitazone on injury of HUVECs induced by homocysteine and the levels of TNF-a and IL-6

目的研究过氧化物酶体增殖剂激活型受体γ(PPARγ)激动剂罗格列酮(Rosiglitazone,Ros)对同型半胱氨酸(homocysteine,Hcy)所致人脐静脉内皮细胞(HUVECs)损伤的保护作用及对肿瘤坏死因子-a(TNF-a)和白介素-6(IL-6)水平的影响。方法将原代培养的HUVECs分为对照组、Hcy刺激组和Ros干预组。MTT检测细胞存活率。免疫细胞化学方法检测Bax、Bcl-2、Caspase-3蛋白表达。ELISA法检测各组细胞培养上清液TNF-a和IL-6水平。Western-blot检测PPARγ蛋白表达。结果与对照组比较,Hcy组细胞存活率明显减少;Bax和Caspase-3蛋白表达上调,Bcl-2和PPARγ蛋白表达下调(P<0.05);TNF-a和IL-6水平增加,而Ros可显著逆转上述变化(P<0.05),并具有时间和剂量依赖性。结论罗格列酮对Hcy所致HBVECs损伤有保护作用(以大剂量组为优),其作用机制与降低TNF-a和IL-6水平有关。

Objective To investigate the protective functions of rosiglitazone（Ros）as a peroxisome proliferator activated receptor agonist（PPARγ）on injury of HUVECs induced by homocysteine（Hcy）and its effect on the levels of TNF-a and interleukin-6.Methods HUVECs by primary culture were divided into three groups：control group,Hcy stimulant group and Rosiglitazone intervention group.Cell viability was measured by MTT assay.The expression of Bax,Bcl-2 and Caspase-3 protein were measured by immunocytochemistry.ELISA was used to examine the levels of TNF-a and IL-6 in cultured supernatants.The expression of PPARγ protein was examined by Western blot.Results Compared with the control group,Cell viability was descended significantly,together with the levels of TNF-a and IL-6 increased clearly,the expression of Bax and Caspase-3 protein was up-regulated,but that of Bcl-2 and PPARγ was down regulated in Hcy group（P〈0.05）.Ros could reverse these changes in a time and concentration-dependent manner（P〈0.05）.Conclusion Rosiglitazone,especially high-dose rosiglitazone,has the protective effect on injury of HUVECs induced by homocysteine,whose mechanism is related to the decrease of the levels of TNF-a and IL-6.