摘要目的 探讨转化生长因子β1(TGF-β1)在大鼠腹膜间皮细胞(RPMC)中的促炎作用及机制.方法 用TGF-β1(10 μg/L)刺激体外培养的RPMC,观察其对RPMC中单核细胞趋化蛋白1(MCP-1)表达的影响.体外转染Smad7和pcDNA3空载体至RPMC后,观察TGF-β1(10 μg/L)对RPMC MCP-1和p38表达的影响.用p38抑制剂SB203580(10 μmol/L)预处理RPMC后,加入TGF-β1(10 μg/L),观察阻断p38信号通路对MCP-1表达的影响.结果 RT-PCR结果显示,3 h、6 h、12 h、24 h TGF-β1刺激组腹膜间皮细胞MCP-1表达均显著高于0 h对照组(P<0.05),6 h刺激组MCP-1表达达高峰(P<0.01).ELISA结果显示,6 h、12 h、24 h、48 h TGF-β1刺激组MCP-1表达增多(P<0.05),48 h表达达高峰(P<0.01).上调表达Smad7和p38抑制剂SB203580都能明显抑制TGF-β1刺激RPMC产生和分泌MCP-1的作用,与pcDNA3空载体组比较,Smad7治疗组MCP-1的表达显著下调(P<0.05);与TGF-β1刺激组比较,SB203580治疗组MCP-1的表达显著下调(P<0.01).TGF-β1能活化p38磷酸化的过程,上调Smad7可抑制TGF-β1对它们的激活反应;与正常对照组比较,TGF-β1刺激组磷酸化(P)-p38的表达显著上调(P<0.05);与TGF-β1刺激组比较,Smad7治疗组p-p38的表达显著下调(P<0.05).结论 TGF-β1能促进RPMC MCP-1的表达,其促炎作用可能是通过激活p38MAPK信号通路介导的.

abstractsObjective To investigate the pro-inflammatory effect of transforming growth factor β1 (TGF-β1) in rat peritoneal mesothelial cells (RPMCs) and its machanism. Methods TGF-β1-induced RPMCs model in vitro was established, and the expression of MCP-1 in the TGF-β1-induced RPMCs was observed. The intervention of Smad7 on the expression of MCP-1 and p38 signal proteins induced by TGF-β1 in RPMCs was explored as well as the intervention of p38 inhibitor SB203580 on the expression of MCP-1 induced by TGF-β1 in RPMCs. Results TGF-β1 could stimulate MCP-1 expression in RPMCs. Compared with control group, MCP-1 mRNA levels were significantly increased after 3 h treatment with TGF-β1 (P<0.05), peak MCP-1 induction occurred at 6 h (P<0.01), and the stimulatory effect of TGF-β1 persisted through 24 h (P<0.05). MCP-1 protein levels were significantly increased after 6 h treatment with TGF-β1(P<0.05), peak MCP-1 induction occurred at 48 h(P<0.01). Over-expressed Smad7 and p38 inhibitor could reduce the expression of MCP-1 induced by TGF-β1 (P<0.05). TGF-β1 could activate p38 signaling pathway, but over-expressed Smad7 could inhibit this role of TGF-β1. Compared with control group, the expression level of p-p38 was increased in TGF-β1-stimulated group. Compared with TGF-β1-stimulated group, the expression level of p-p38 was reduced in Smad7 gene transfer group. Conclusions TGF-β1-induced MCP-1 expression in rat peritoneal mesothelial cells is p38MAPK dependent.