摘要
目的 观察转化生长因子β1（TGF-β1）对脂多糖（LPS）刺激大鼠腹膜间皮细胞上调表达促炎症因子的影响，并探讨其可能的机制。方法 把原代培养的第2代大鼠腹膜间皮细胞（RPMCs）分成对照组、LPS刺激组（1mg／L）、TGF-β1刺激组（5μg／L）及LPS＋TGF-β1刺激组。RT—PCR和ELISA方法检测肿瘤坏死因子α（TNF-α）和白细胞介素6（IL-6）mRNA及蛋白的表达。蛋白印迹方法检测磷酸化核因子（p-NF）-κB／NF—κB值的变化。结果 （1）LPS可刺激RPMCs上调TNF—α和IL-6表达。LPS刺激24h后，p—NF—κB／NF—κB值升高。（2）TGF—β1可拮抗LPS刺激大鼠腹膜间皮细胞上调TNF-α和IL-6表达，同时，也降低p—NF—κB／NF—κB值。结论 在体外培养的大鼠腹膜间皮细胞，TGF-β1可拮抗LPS的致炎作用，其作用机制可能是通过抑制NF—κB的活化而介导。
Objective To observe the effect of TGF-β1 on the pro-inflammatory cytokine expression induced by lipopolysaccharide （LPS） in rat peritoneal mesothelial cells （RPMCs）, and explore its possible mechanism. Methods RPMCs were isolated from rat peritoneum. The RPMCs were incubated with LPS （1 mg/L）, TGF-β1（5 μg/L）, or stimulated by LPS （1 mg/L） after incubated with TGF-β1 （5 μg/L） for 1 h. RPMCs of control group were just incubated with medium. TNF-α and IL-6 mRNA were detected by RT-PCR. Their protein levels were examined by ELISA. NF-κB protein was detected by Western blot. Results Compared with control group, the expression of pro-inflammatory cytokine （TNF-α, IL-6） was significantly increased in the groups stimulated by LPS and TGF-β1. Also, the NF-κB was activated by LPS and TGF-β1. In the group pre-incubated by TGF-β1 for 1 hour, up-regulating expression of TNF-α mRNA and protein was significantly inhibited. Furthermore, NF-κB activation was also partly inhibited by pre-treatment with TGF-β1. Conclusion TGF-β1 may inhibit pro-inflammatory cytokine expression induced by lipopolysaccharide in rat peritoneal mesothelial cells through the inactivation of NF-κB signal transduction pathway.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2007年第3期194-198,共5页
Chinese Journal of Nephrology
关键词
脂多糖类
转化生长因子β
NF—κβ
间皮细胞
腹膜炎
Lipopolysaccharides
Transforming growth factor beta
NF-kappa B
Mesothelial cells
Peritonitis
