Jiang Tao, Che Qi, Zhao Zhong-hua, Zhang Xiu-rong, Zhang Nong
Department of Pathology, Shanghai Medical College, Fudan University, Shanghai 200032, China.
Zhonghua Bing Li Xue Za Zhi. 2005 Mar;34(3):171-4.
To study the effect of aldose reductase (AR) on expression of fibronectin and collagen IV in cultured rat renal mesangial cells (MsC).
AR expression plasmid vector (pCDNA3-AR) was constructed by restriction endonuclease digestion and ligation procedures. Stable expression of AR in MsC was established by Lipofectin transfection. Western blot and immunofluorescence analyses were performed to verify the transfection efficiency. Expression of fibronectin and collagen IV proteins were analyzed using Western blot.
Expression of fibronectin and collagen IV in naive MsC treated with TGF-beta1 was upregulated in comparison to that of the untreated naive MsC (P < 0.01). MsC transfected with pCDNA3-AR showed a remarkable increase of expression of fibronectin and collagen IV (P < 0.01). Aldose reductase inhibitors (Sorbinil and Zopolrestat) significantly inhibited the expression of fibronectin and collagen IV in naive MsC (P < 0.05).
Overexpression or inhibition of AR activity significantly alters the expression of fibronectin and collagen IV proteins in cultured rat MsC, suggesting that AR plays a significant role in the pathogenesis of glomerulosclersis.
研究醛糖还原酶（AR）对培养的大鼠肾系膜细胞（MsC）中纤连蛋白和IV型胶原表达的影响。
通过限制性内切酶消化和连接程序构建AR表达质粒载体（pCDNA3-AR）。采用脂质体转染法在MsC中建立AR的稳定表达。进行蛋白质免疫印迹和免疫荧光分析以验证转染效率。采用蛋白质免疫印迹法分析纤连蛋白和IV型胶原蛋白的表达。
与未处理的正常MsC相比，用转化生长因子-β1（TGF-β1）处理的正常MsC中纤连蛋白和IV型胶原的表达上调（P<0.01）。用pCDNA3-AR转染的MsC显示纤连蛋白和IV型胶原的表达显著增加（P<0.01）。醛糖还原酶抑制剂（索比尼尔和唑泊司他）显著抑制正常MsC中纤连蛋白和IV型胶原的表达（P<0.05）。
AR活性的过表达或抑制显著改变培养的大鼠MsC中纤连蛋白和IV型胶原蛋白的表达，提示AR在肾小球硬化的发病机制中起重要作用。
