摘要目的 探讨高糖环境下球形脂联素(globular adiponectin,gAd)对大鼠近端肾小管上皮细胞(NRK-52E)单核趋化蛋白-1 (MCP-1)表达的影响以及其与脂联素受体(AdipoR)、p38丝裂原活化蛋白激酶(p38MAPK)的关系.方法 体外培养NRK-52E细胞,分为六组:正常对照组(NG,5.6 mmol/L葡萄糖)、高糖组(HG,25 mmol/L葡萄糖)、gAd处理组1～3(gAd 1～3,分别用2 mg/L、5 mg/L、10 mg/L gAd+25 mmol/L葡萄糖)、p38MAPK抑制剂组(SB,10 μmol/LSB203580+25 mmol/L葡萄糖).分别采用RT-PCR和Real-time PCR法检测MCP-1和AdipoR1/AdipoR2的mRNA表达;Western印迹法检测磷酸化p38MAPK(p-p38 MAPK)、总p38MAPK(t-p38 MAPK)、MCP-1和AdipoR1/AdipoR2的蛋白表达.结果 与NG组相比,HG组MCP-1的mRNA和蛋白表达、p-p38MAPK蛋白表达均明显升高(均P＜0.05),t-p38 MAPK无明显变化.gAd处理组及p38MAPK抑制剂组p-p38MAPK、MCP-1的mRNA和蛋白表达明显降低(均P＜0.05).AdipoR1及AdipoR2在NG组均有表达,且AdipoR1的mRNA和蛋白表达均明显高于AdipoR2,差异有统计学意义(均P＜ 0.01).与NG组比较,HG组及AdipoR2的mRNA和蛋白表达稍低,但差异均无统计学意义(P＞0.05).与HG组比较,gAD处理组AdipoR1的mRNA和蛋白表达均增加(均P＜ 0.01),但AdipoR2的mRNA及蛋白表达无改变(P＞0.05).结论 gAd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的高表达,且这种肾脏保护作用是AdipoR1及p38MAPK介导的.

abstractsObjective To investigate the effect of globular adiponectin on the high expression of monocyte chemotactic protein-1 (MCP-1) induced by high glucose in rat renal tubular epithelial cells (NRK52E),and its relationship with adiponectin receptors and p38MAPK.Methods NRK52E cells were cultured in vitro and divided into six groups:normal glucose group (NG,5.6 mmol/L glucose),high glucose group(HG,25 mmol/L glucose),gAd groupl (HG+gAd 2 mg/L),gAd group2 (HG+gAd 5 mg/L),gAd group3 (HG+gAd 10 mg/L),p38MAPK antagonist group:(SB,HG+SB203580 10 μmol/L).The protein expression of phosphorylated p38MAPK (p-p38MAPK),total p38MAPK (t-p38MAPK),MCP-1 and AdipoR1/AdipoR2 were examined by western blotting.The mRNA expression of MCP-1 and AdipoR1/AdipoR2 were detected by RT-PCR and real-time PCR respectively.Results Compared with NG group,the mRNA and protein expression of MCP-1 increased significantly in HG group (all P＜ 0.05).The phosphorylation of p38MAPK increased (P＜ 0.05) with no change in t-p38MAPK protein.The addition of gAd or SB203580 inhibited the unregulation of MCP-1 and p-p38MAPK induced by HG.Two kinds of adipoR,adipoR1 and adipoR2,were all detectable in NG group,and mRNA and protein expression of adipoR1 was higher than that of adipoR2 (P＜ 0.01).Compared with NG group,the expression of adipoR decreased in HG group,but the difference had no statistical significance(P ＞ 0.05).Compared to HG group,the mRNA and protein expression of adipoR1 increased in gAd groups (all P ＜ 0.01).Conclusion The gAd can dose-dependently attenuate the overexpression of MCP-1 induced by high glucose,and this protective effect may be mediated by adipoR1 and p38MAPK.