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目的:探讨C反应蛋白(CRP)对肾小管上皮细胞NF-κB信号与IL-6 mRNA表达的影响。方法:体外培养的骨小管上皮细胞NRK-52E,分3组:①对照组:设阴性对照(加入PBS)和阳性对照(AngⅡ,1μmol/L);②CRP刺激组(分别加入CRP 5,10,20 mg/L);③干预组:加入10 mg/L CRP,同时加入CRP抗体1μmol/L或SB203580(10μmol/L,p38MAPK的特异性抑制剂)。分别应用ELISA、RT-PCR、Western Blot技术和EMSA方法,检测细胞p38MAPK的磷酸化、NF-κB活性与IL-6分泌与mRNA表达的变化。结果:CRP呈剂量依赖性上调NRK-52E细胞IL-6分泌与mRNA的表达。CRP上调NRK-52E细胞p38MAPK的磷酸化与NF-κB与DNA的结合活性。CRP抗体、SB203580下调CRP对NRK-52E细胞的炎性激活效应。结论:CRP直接诱导NRK-52E细胞NF-κB活化及下游因子IL-6 mRNA与蛋白质的高表达,可能通过p38MAPK的磷酸化途径。
Objective: To investigate the effect of C-reactive protein (CRP) on the expression of NF-κB signal and IL-6 mRNA in renal tubular epithelial cells. Methods: Rabbit epithelial cells NRK-52E cultured in vitro were divided into 3 groups: ① control group: negative control (PBS) and positive control (AngⅡ, 1μmol / L); ② CRP stimulation group 20 mg / L). ③Intervention group: 10 mg / L CRP was added, and CRP antibody 1μmol / L or SB203580 (10μmol / L, a specific inhibitor of p38MAPK) was added at the same time. The changes of p38MAPK phosphorylation, NF-κB activity and IL-6 secretion and mRNA expression were detected by ELISA, RT-PCR, Western Blot and EMSA respectively. Results: CRP up-regulated IL-6 secretion and mRNA expression in NRK-52E cells in a dose-dependent manner. CRP up-regulates the phosphorylation of p38 MAPK and the binding activity of NF-κB to DNA in NRK-52E cells. CRP antibody, SB203580 down-regulates the inflammatory activation effect of CRP on NRK-52E cells. CONCLUSION: CRP directly induces the activation of NF-κB and down-regulates the expression of IL-6 mRNA and protein in NRK-52E cells, possibly through the phosphorylation pathway of p38MAPK.