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目的探讨转化生长因子β(TGF-β)/Smad信号通路对肾小球系膜细胞环氧化酶2(COX-2)表达的影响。方法取传代培养第4代系膜细胞进行分组:对照组(A组)、TGF-β5 ng/ml组(B组)、TGF-β10 ng/ml组(C组)、TGF-β10 ng/ml+NS-398 10μmol(D组)和TGF-β10 ng/ml+staurosporine 5 nmol/ml组(E组)。用RT-PCR方法测定各组COX-2 mRNA及Smad 2 mRNA的表达,Western blot法测定各组COX-2及Smad 2蛋白的表达。结果不同浓度TGF-β作用4、12、24 h后,系膜细胞Smad 2 mRNA和COX-2 mRNA表达及二者的蛋白表达均增加,且呈浓度和时间依赖性(P<0.05),加入COX-2抑制剂NS-398或Smad 2抑制剂staurosporine后,COX-2和Smad 2的表达均减少(P<0.05)。COX-2和Smad 2表达呈正相关(r=0.9438,P<0.05)。结论 TGF-β可通过Smad信号通路刺激肾小球系膜细胞COX-2的表达。
Objective To investigate the effect of transforming growth factor β (TGF-β) / Smad signaling pathway on the expression of cyclooxygenase 2 (COX-2) in glomerular mesangial cells. Methods The mesangial cells of passage 4 were subcultured and divided into control group (group A), TGF-β5 ng / ml group (group B), TGF-β10 ng / ml group + NS-398 10 μmol (group D) and TGF-β 10 ng / ml + staurosporine 5 nmol / ml group (group E). The expressions of COX-2 mRNA and Smad 2 mRNA in each group were determined by RT-PCR. The expressions of COX-2 and Smad 2 in each group were determined by Western blot. Results The expressions of Smad 2 mRNA and COX-2 mRNA in mesangial cells were increased at different concentrations of TGF-β for 4, 12 and 24 h, both in a concentration-and time-dependent manner (P <0.05) After COX-2 inhibitor NS-398 or Smad 2 inhibitor staurosporine, the expression of COX-2 and Smad 2 were decreased (P <0.05). There was a positive correlation between the expression of COX-2 and Smad 2 (r = 0.9438, P <0.05). Conclusion TGF-β can stimulate the expression of COX-2 in mesangial cells through Smad signaling pathway.