论文部分内容阅读
目的探讨在大鼠腹膜间皮细胞(RPMC)中转化生长因子β1(TGF-β1)对肿瘤坏死因子-α(tumor necrosisfactor-α,TNF-α)表达的影响及其机制。方法在体外培养的大鼠腹膜间皮细胞模型中,加入TGF-β1(10ng/ml)预刺激,研究RPMC中肿瘤坏死因子-α(TNF-α)的表达;体外转染Smad7至RPMC后,研究其对TGF-β1(10 ng/ml)刺激后RPMCs中TNF-α和p38表达的影响。p38抑制剂SB203580(10umol/L)预处理RPMCs后,加入TGF-β1(10 ng/ml)刺激,研究抑制p38信号通路后对TNF-α表达的影响。结果RT-PCR结果显示,与0h对照组相比,3 h、6 h、12 h TGF-β1刺激组TNF-α表达明显增加(P<0.05)。上调表达Smad7抑制TGF-β1刺激RPMC产生TNF-α的作用,p38抑制剂SB203580亦能抑制TGF-β1刺激RPMC表达TNF-α的作用。TGF-β1参与p38磷酸化的活化,Smad7可抑制TGF-β1对它的激活反应;与正常对照组比较,TGF-β1刺激组磷酸化(p)-p38的表达显著增加(P<0.05),与TGF-β1刺激组比较,Smad7治疗组p-p38的表达显著减弱(P<0.05)。结论在大鼠腹膜间皮细胞中,TGF-β1能促进TNF-α的表达,其作用机制可能是依赖活化p38MAPK信号通路来实现的。
Objective To investigate the effect of transforming growth factor-β1 (TGF-β1) on the expression of tumor necrosis factor-α (TNF-α) and its mechanism in rat peritoneal mesothelial cells (RPMC) Methods The expression of tumor necrosis factor-α (TNF-α) in RPMC was determined by adding TGF-β1 (10 ng / ml) to rat peritoneal mesothelial cells in vitro. After transfection of Smad7 into RPMC, To investigate the effect of TGF-β1 (10 ng / ml) on the expression of TNF-α and p38 in RPMCs. After pretreatment with p20 inhibitor SB203580 (10umol / L), RPMCs were stimulated with TGF-β1 (10 ng / ml) to study the effect of p38 signaling pathway on TNF-α expression. Results The results of RT-PCR showed that the expression of TNF-α in TGF-β1 stimulation group at 3 h, 6 h and 12 h increased significantly compared with 0 h control group (P <0.05). Upregulation of Smad7 inhibited TGF-β1-induced RPMC production of TNF-α, and p38 inhibitor SB203580 also inhibited TGF-β1-induced RPMC expression of TNF-α. TGF-β1 is involved in the activation of p38 phosphorylation, and Smad7 inhibits the activation of TGF-β1. Compared with the normal control group, the phosphorylation of p38-p38 was significantly increased (P <0.05) Compared with the TGF-β1-stimulated group, the expression of p-p38 in Smad7-treated group was significantly decreased (P <0.05). Conclusion TGF-β1 can promote the expression of TNF-α in rat peritoneal mesothelial cells, and its mechanism may be dependent on activation of p38MAPK signaling pathway.