热休克蛋白72肽结合区对肾小管上皮间质转分化的影响

中华肾脏病杂志 ›› 2012, Vol. 28 ›› Issue (6) : 484-488.

基础研究

曹陶曹师荣李辉雁熊丽萍范瑾瑾余学清毛海萍

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Impact of peptide binding domain of heat shock protein 72 on epithelial to mesenchymal transition

CAO Tao, CAO Shi-rong, LI Hui-yan, XIONG Li-ping, FAN Jin-jin, YU Xue-qing, MAO Hai-ping.

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摘要

目的探讨热休克蛋白72肽结合区在肾小管上皮间质转分化（EMT）过程中的作用和可能机制。方法应用质粒转染方法分别诱导热休克蛋白72（HSP72）野生型、肽结合区缺失型（HSP72-△PBD）和肽结合区（PBD）的表达。用转化生长因子β1（TGF-β1）刺激大鼠肾小管上皮细胞（NRK-52E）48 h，Western 印迹和免疫荧光染色检测细胞E-钙黏蛋白（cadherin），α-平滑肌肌动蛋白（SMA），HSP72和Smad3/磷酸化（p）-Smad3蛋白表达。结果TGF-β1（10 μg/L）刺激NRK-52E细胞48 h后上调α-SMA和下调 E-cadherin蛋白表达水平。Western印迹及细胞免疫荧光显示，过表达HSP72和PBD能明显减轻TGF-β1诱导的NRK-52E细胞E-cadherin蛋白表达下调和α-SMA蛋白表达上调，而过表达HSP72-△PBD不能改变上述蛋白的表达。此外，过表达HSP72和PBD显著抑制Smad3的磷酸化。结论 HSP72抑制Smad3活化和EMT的发生可能与PBD的功能有关。

Abstract

Objective To investigate the effects of peptide-binding domain (PBD) of heat shock protein (HSP) 72 on epithelial to mesenchymal transition (EMT) in rat renal tubular epithelial cells. Methods The expressions of wild-type HSP72, mutant of HSP72 lacking peptide binding domain (HSP72 -△PBD) and HSP72-PBD were induced by plasmid transfection. NRK-52E cells were stimulated by TGF-β1 for 48 h. The expressions of α-smooth muscle actin (α-SMA), E-cadherin, HSP72 and Smad3/p-Smad3 were detected by Western blot and immunofluorescence. Results After NRK-52E cells were stimulated by TGF-β 1 (10 μg/L) for 48 h, the expression of α-SMA was increased and the protein level of E-cadherin was decreased. Western blotting and immunofluorescence showed that over-expression of both HSP72 and PBD inhibited TGF-β1-induced up-regulation of protein α-SMA expression, down-regulation of protein E-cadherin. However, over-expression of HSP72-△PBD did not change the protein level of E-cadherin and α-SMA. In addition, over-expression of HSP72 and PBD significantly inhibited the phosphorylation of Smad3. Conclusion Inhibition of Smad3 activation and EMT by HSP72 is associated with the function of PBD.

导出引用

曹陶曹师荣李辉雁熊丽萍范瑾瑾余学清毛海萍. 热休克蛋白72肽结合区对肾小管上皮间质转分化的影响[J]. 中华肾脏病杂志, 2012, 28(6): 484-488.

CAO Tao;CAO Shi-rong;LI Hui-yan;XIONG Li-ping;FAN Jin-jin;YU Xue-qing;MAO Hai-ping. . Impact of peptide binding domain of heat shock protein 72 on epithelial to mesenchymal transition[J]. Chinese Journal of Nephrology, 2012, 28(6): 484-488.

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Received	Revised	Published
1900-01-01	1900-01-01	2012-06-15
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2013-01-31

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