Huang Ping, Zhang Yue-juan, Huang Yuan, Zhao Jing-jing, Jiang Tao, Zhang Nong
Department of Pathology, Shanghai Medical College, Fudan University, Shanghai 200032, China.
Zhonghua Bing Li Xue Za Zhi. 2010 Jun;39(6):405-9.
To study the effect of PKC signalling pathway and aldose reductase (AR) on the expression of fibronectin (FN) induced by transforming growth factor-β1 (TGF-β1).
Human mesangial cells (HMCs) were cultured and transfected with pcDNA3-AR, and subject to AR gene silencing with small interfering RNA (siRNA) and then the cell was treated with recombinant human TGF-β1. The AR mRNA expression in the HMCs was examined using real time RT-PCR and protein expression of AR and FN was detected by Western blotting.
The cultured HMC treated with TGF-β1 showed increased expression of AR and FN, the normal HMC showed not reduced expression of FN after incubation with single inhibitors of AR.Pre-incubation of cells with inhibitors of AR and PKC, then the different groups of cells were treated with TGF-β1, and the induction effect on FN expression was suppressed (34%) in HMC. HMCs transfected with AR showed a strong protein expression of FN, which was increased by 3.6-fold after treatment with TGF-β1 (P < 0.05), and the induction effect on FN expression was suppressed by GÖ6983 (42%) in HMCs (P < 0.05). The HMC with AR gene knock-down by siRNA showed a decreased expression of AR and 90% decrease of FN protein in HMCs (P < 0.01), and TGF-β1-induced up-regulation of FN was significantly suppressed by siRNA (12%) in HMCs (P < 0.01).
AR is capable of regulating FN expression only in the presence of TGF-β1, and this reaction is possibly accomplished through the activation of PKC signalling pathway.
研究蛋白激酶C（PKC）信号通路和醛糖还原酶（AR）对转化生长因子-β1（TGF-β1）诱导的纤连蛋白（FN）表达的影响。
培养人肾小球系膜细胞（HMCs），用pcDNA3-AR转染，并用小干扰RNA（siRNA）进行AR基因沉默，然后用重组人TGF-β1处理细胞。采用实时逆转录聚合酶链反应（RT-PCR）检测HMCs中AR mRNA的表达，用蛋白质印迹法检测AR和FN的蛋白表达。
用TGF-β1处理的培养HMC显示AR和FN的表达增加，正常HMC在用AR单一抑制剂孵育后FN表达未降低。细胞先用AR和PKC抑制剂预孵育，然后用TGF-β1处理不同组细胞，对FN表达的诱导作用在HMC中受到抑制（34%）。转染AR的HMC显示FN蛋白表达强烈，用TGF-β1处理后增加了3.6倍（P<0.05），GÖ6983对HMC中FN表达的诱导作用受到抑制（42%）（P<0.05）。用siRNA敲低AR基因的HMC显示AR表达降低，HMC中FN蛋白降低90%（P<0.01），siRNA对TGF-β1诱导的HMC中FN上调有显著抑制作用（12%）（P<0.01）。
AR仅在TGF-β1存在时能够调节FN表达，且该反应可能通过PKC信号通路的激活来完成。
