锌离子对大鼠腹膜间皮细胞转分化及TGFβ/Smad通路的影响  

Zinc Ion Protects Rat Peritoneal Mesothelial Cells from Undergoing Epithelial-to-mesenchymal Transition via Inhibiting TGF-β/Smad Pathways

作　　者：张秀丽[1,2] 梁单[3] 李红娟[3] 马健飞[1] 胡晔[1] 樊怡[1] 王力宁[1] 

机构地区：[1]中国医科大学第一附属医院肾内科 [2]本溪市中心医院肾内科 [3]中国人民解放军95935部队

出　　处：《中国医科大学学报》2013年第11期1003-1007,共5页Journal of China Medical University

摘　　要：目的研究锌离子对脂多糖(LPS)诱导的大鼠腹膜间皮细胞(RPMCs)转分化(EMT)的影响,为锌离子在腹膜透析中的应用提供理论基础。方法胰蛋白酶消化法培养原代大鼠腹膜间皮细胞、传代,经鉴定后分组:(1)对照组,(2)LPS组,(3)ZnSO4组,(4)ZnSO4/LPS组。应用Western blot方法、免疫荧光方法分别检测α平滑肌肌动蛋白(αSMA)、上皮钙黏素(E cadherin)、Ⅰ型胶原(collagenⅠ)、Smad2、Smad3、磷酸化的Smad2、3(P Smad2、3)蛋白表达;酶联免疫吸附方法检测上清液TNFα、TGFβ1的表达。结果与LPS组相比,ZnSO4/LPS组的αSMA的免疫荧光强度减弱;与对照组相比,LPS组RPMC的αSMA、collagenⅠ蛋白表达明显升高,而E cadherin蛋白表达降低。与LPS组相比,ZnSO4/LPS组RPMC的αSMA、collagenⅠ表达明显降低、E cadherin蛋白表达增多;LPS组上清液中TNFα、TGFβ1浓度显著高于对照组;ZnSO4/LPS组上清液中TNFα、TGFβ1浓度显著低于LPS组;与LPS组相比,ZnSO4/LPS组P Smad2、P Smad3的表达明显降低。结论 ZnSO4可能可以逆转LPS所致的RPMC的EMT,对腹膜透析过程中所导致的RPMCs损伤可能具有保护作用,其作用机制可能是通过抑制TGFβ/Smad信号传导通路。Objective To investigate the effects of Zn ion on lipopolysaccharide （LPS）-induced epithelial-to-mesenchymal transition （EMT） in rat peritoneal mesothelial cells （RPMCs） and explore the related molecular mechanisms. Methods RPMCs were isolated, cultured and passaged by enzymatic disaggregation, then identified with immtmocytochemistry method under phase contrast inverted microscope and transmission electron microscope. RPMCs were incubated with 100 μmol/L LPS for 48 hours, or pre-treated with 30 μmol/L ZnSO4 for 24 hours followed incubated with 100 μmol/L LPS for 48 hours. RPMCs in the control group were just incubated with medium. The expression of α-SMA, E-cadherin, collagen I, Smad2, Smad 3, P-Smad2, and P-Smad 3 was detected by Western Blot or Innnunofluorescence staining. In addition, Elisa analysis was performed to investigate the change of TNF-α, TGF-β1 in the culture medium. Results We found that Zn ion supplementation significantly inhibited LPS-in- duced RPMC EMT, by attenuating TNF-α, TGF-β1 production. The LPS-induced EMT can be attenuated by pre-treating the RPMCs with 30 μmol/ L ZnSO4, resulted in reduced up-regulation of α-SMA and mesenchymal marker collagen I, and ameliorated expression of epithelial protein E-cad- herin. Further analysis revealed that Zn supplementation inhibited LPS- induced RPMC EMT likely through inhibition of TGF- β/Smad pathways. Conclusion These results indicate that ZnSO4 can inhibit EMT in LPS-induced RPMCs lhrough inhibition of TNF-α, TGF-β1 production as well as TGF-β/Smad pathways activation.

关 键 词：脂多糖 转分化 大鼠腹膜间皮细胞 腹膜透析 

分 类 号：R572.2[医药卫生—消化系统]