中国医科大学学报

中国医科大学学报

中国医科大学学报 ›› 2013, Vol. 42 ›› Issue (11): 1003–1004.

• 论著 • 上一篇    下一篇

锌离子对大鼠腹膜间皮细胞转分化TGF-β/Smad通路的影响

张秀丽1,2,梁单3,马健飞1,樊怡1,王力宁1   

  1. 1.中国医科大学第一附属医院肾内科,沈阳,110001;
    2.本溪市中心医院肾内科,本溪,117000;
    3.中国人民解放军95935部队,哈尔滨,150000
  • 出版日期:2013-11-30 发布日期:2013-11-28
  • 作者简介:张秀丽(1972-),女,博士,副主任医师。联系电话:18841475918

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

ZHANG Xiu-li1,2,LANG Dan 3,MA Jian-fei1,FAN Yi1,WANG Lin-ing1   

  1. 1.Department of Nephrology,The First Hospital,China Medical University,Shenyang 110001,China;
    2.Department of Nephrology,Benxi Centre Hospital,Benxi 117000,China;
    3. Troops of 95935 Unit PR China,Haerbin 150000,China
  • Online:2013-11-30 Published:2013-11-28

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Westernblot方法、免疫荧光方法分别检测#x003b1;平滑肌肌动蛋白(#x003b1;-SMA)、上皮钙黏素(E-cadherin)、#x02160;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(ELISA)检测上清液TNF-#x003b1;、TGF-#x003b2;1的表达。结果与LPS组相比,ZnSO4/LPS组的#x003b1;-SMA的免疫荧光强度减弱;与对照组相比,LPS组RPMC的#x003b1;-SMA、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#x003b1;-SMA、collagenI表达明显降低、E-cadherin蛋白表达增多;LPS组上清液中TNF-#x003b1;、TGF-#x003b2;1浓度显著高于对照组;ZnSO4/LPS组上清液中TNF-#x003b1;、TGF-#x003b2;1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TGF-#x003b2;/Smad信号转导通路。

关键词: 脂多糖, ZnSO4, 转分化, 大鼠腹膜间皮细胞, 腹膜透析

Abstract: 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 immunocytochemistry method under phase contrast inverted microscope and transmission electron microscope. RPMCs were incubated with 100#x003bc;M LPS for 48 hours,or pre-treated with 30#x003bc;M ZnSO4 for 24 hours followed incubated with 100#x003bc;M LPS for 48 hours. RPMCs in the control group were just incubated with medium. The expression of #x003b1;-SMA ,E-cadherin ,collagen I,Smad2,Smad 3 ,P-Smad2,and P-Smad 3 was detected by Western Blot or Immuno?uorescence staining. In addition,Elisa analysis was performed to investigate the change of TNF-#x003b1;,TGF-#x003b2;1 in the culture medium.Results We found that Zn ion supplementation significantly inhibited LPS-induced RPMC EMT,by attenuating TNF-#x003b1;,TGF-#x003b2;1 production. The LPS-induced EMT can be attenuated by pre-treating the RPMCs with 30#x003bc;M ZnSO 4, resulted in reduced up-regulation of #x003b1;-SMA and mesenchymal marker collagen I,and ameliorated expression of epithelial protein E-cadherin. Further analysis revealed that Zn supplementation inhibited LPS-induced RPMC EMT likely through inhibition of TGF-#x003b2;/Smad pathways.Conclusion These results indicate that ZnSO4 can inhibit EMT in LPS-induced RPMCs through inhibition of TNF-#x003b1;,TGF-#x003b2;1 production as well as TGF-#x003b2;/Smad pathways activation.

Key words: lipopolysaccharide, ZnSO epithelial-to-mesenchymal transition

中图分类号: 

  • R572.2
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