球形脂联素对高糖诱导的NRK-52E细胞单核趋化蛋白-1的表达影响及其可能机制

Effects of Global Adiponectin on the Expression of Monocyte Chemoattractant Protein-1 in NRK-52E Cells Induced by High Glucose and Its Possible Mechanisms

目的:探讨球形脂联素(gAd)对高糖环境下大鼠近端肾小管上皮细胞(NRK-52E)单核趋化蛋白-1(MCP-1)mRNA和蛋白表达的影响以及其与p38丝裂原活化蛋白激酶(p38MAPK)通路的关系。方法:将体外培养NRK-52E细胞分为6组:正常糖对照组NG(含5.6mmol/L葡萄糖)、高糖组HG(含25mmol/L葡萄糖)、gAd处理组(分别用gAd2,5,10mg/L+25mmol/L葡萄糖)、p38MAPK抑制剂组(25mmol/L葡萄糖+10μmol/LSB203580)。30min后采用Western印迹方法检测磷酸化p38MAPK(p-p38MAPK)和总p38MAPK(t-p38MAPK)的表达;24hRT-PCR法、Western印迹方法分别检测MCP-1mRNA和蛋白的表达。结果:高糖环境下,NRK-52E细胞p-p38MAPK、MCP-1mRNA和蛋白表达明显升高(P<0.05),加入gAd及SB203580后,NRK-52E细胞p-p38MAPK、MCP-1mRNA和蛋白表达明显降低(P<0.05),且呈现剂量依赖性(P<0.05)。结论:gAd呈现剂量依赖性抑制高糖诱导的大鼠近端肾小管上皮细胞MCP-1高表达,且这种肾脏保护作用是通过p38MAPK途径介导的。

Objective：To investigate the effect of globular adiponectin（gAd）on the expression of monocyte chemoattractant protein-1（MCP-1）in rat renal tubular epithelial cells（NRK-52E）induced by high glucose and the function of p38MAPK pathway.Methods：NRK-52Ecells were cultured in vitro and divided into six groups：normal glucos（NG,5.6mmol/L）,high glucose group（HG,25mmol/L）,HG＋2,5,and 10mg/L gAd groups,and HG ＋ p38MAPK antagonist group（SB203580,10μmol/L）.Thirty minutes later,the expression of p-p38MAPK and t-p38MAPK were examined by Western blotting.After 24h,RT-PCR and Western blot were used to determine the expression of MCP-1.Results：Compared with that in NG group,the expression of pp38MAPK and MCP-1increased significantly after high glucose treatment,but gAd at different concentrations and SB203580could inhibit these changes.Conclusion：Globular adiponectin can attenuate the over expression of MCP-1induced by high glucose dose-dependently and this protective effect is mediated by p38MAPK signaling pathway.