转化生长因子β1对大鼠腹膜间皮细胞IL-8表达的影响

Effect of TGF-β1 on Interleukin-8 Expression in Rat Peritoneal Mesothelial Cells

目的探讨转化生长因子β1(TGF-β1)对大鼠腹膜间皮细胞(RPMCs)IL-8表达的影响及机制。方法 10μg.L-1 TGF-β1刺激体外培养的RPMCs,RT-PCR法检测IL-8的表达;体外转染Smad7和pcDNA3空载体至RPMCs后,观察10μg.L-1 TGF-β1刺激RPMCs对IL-8和p38表达的影响。采用p38抑制剂SB203580(10μmol.L-1)预处理RPMCs后,加入10μg.L-1 TGF-β1,观察阻断p38信号通路对IL-8表达的影响。结果与0h刺激组相比,3、6、12hTGF-β1刺激组IL-8表达明显增加(P<0.05或P<0.01),其中3h达高峰。上调表达Smad7以及SB203580均能显著抑制TGF-β1刺激RPMCs产生IL-8(P<0.01)。与正常对照组比较,TGF-β1刺激磷酸化p38(p-p38)的表达显著增加(P<0.05),与TGF-β1刺激组比较,外源性Smad7转染组p-p38的表达显著减弱(P<0.05)。结论 TGF-β1促进RPMCs IL-8的表达,其机制可能是TGF-β1通过活化p38磷酸化来实现的。

Objective To observe the effect of TGF-β1 on interleukin-8 expression in rat peritoneal mesothelial cells（RPMCs）,and to investigate its mechanism of action. Methods RPMCs were treated with TGF-β1（10 μg·L-1） in vitro and IL-8 expression in RPMCs was detected by RT-PCR.The effect of TGF-β1 on the expression of IL-8 and p38 was investigated after RPMCs were transfected with Smad7 expression plasmid and empty pcDNA3 vector.After pretreatment with p38 inhibitor SB203580（10 μmol·L-1）,RPMCs were stimulated with TGF-β1 to observe the influence of the inhibition of p38 signaling pathway on the expression of IL-8. Results Comparing with control group,the expression of IL-8 increased significantly after treatment with TGF-β1 for 3,6 and 12 hours（P＆lt;0.05 or P＆lt;0.01）.The peak IL-8 expression occurred after treatment for 3 hours（P＆lt;0.05）.Both Smad7 overexpression and SB203580 treatment significantly inhibited IL-8 expression induced by TGF-β1（P＆lt;0.01）.Compared with control group,the expression of p-p38 was significantly increased by TGF-β1 treatment（P＆lt;0.05）.Compared with TGF-β1 tretment group,the expression of p-p38 was significantly reduced by Smad7 transfection.Conclusion TGF-β1 can promote IL-8 expression in RPMCs through inducing p38 phosphorylation.