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/L SB203580+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。
ObjectiveTo 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.
MethodsNRK52E 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 group1 (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.
ResultsCompared 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).
ConclusionThe 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.
姚涛,吴小燕,於文丽,等. 球形脂联素抑制高糖环境下肾小管上皮细胞单核趋化蛋白-1的高表达[J]. 中华肾脏病杂志,2015,31(2)：145-149.
DOI:10.3760/cma.j.issn.1001-7097.2015.02.012版权归中华医学会所有。
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