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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 globulin (gAd) on mRNA and protein expression of monocyte chemoattractant protein-1 (MCP-1) in proximal tubular epithelial cells (NRK-52E) Mitogen-activated protein kinase (p38MAPK) pathway. Methods: NRK-52E cells were divided into 6 groups: NG (5.6 mmol / L glucose), HG (25 mmol / L glucose), gAd treatment group 10mg / L + 25mmol / L glucose), p38MAPK inhibitor group (25mmol / L glucose + 10μmol / LSB203580). The expression of p-p38MAPK (p-p38MAPK) and total p38MAPK (t-p38MAPK) were detected by Western blotting after 30min. The expression of MCP-1mRNA and protein were detected by RT-PCR and Western blotting respectively. Results: The expressions of p-p38MAPK and MCP-1 mRNA and protein in NRK-52E cells were significantly increased under high glucose conditions (P <0.05). After addition of gAd and SB203580, the expressions of p-p38MAPK and MCP- (P <0.05), and showed a dose-dependent manner (P <0.05). CONCLUSION: GAd exhibits a dose-dependent inhibition of high glucose-induced MCP-1 expression in rat proximal tubular epithelial cells, and this renal protective effect is mediated through the p38MAPK pathway.