ClC-3氯通道参与TGF-β通路调控PTH的促成骨分化作用机制研究

›› 2016, Vol. 7 ›› Issue (1): 1-5.

• 论著 • 下一篇

ClC-3氯通道参与TGF-β通路调控PTH的促成骨分化作用机制研究

张瑞¹,王欢¹,刘竹影¹,沈鹏²,丁寅¹

1. 第四军医大学口腔医院正畸科
2. 第四军医大学口腔医院修复科

收稿日期:2015-11-20 修回日期:2015-12-19 出版日期:2016-03-25 发布日期:2016-04-06
通讯作者: 丁寅 E-mail:dingyin@fmmu.edu.cn
基金资助:
国家自然科学基金青年科学基金项目

ClC-3 chloride channel in the regulation of TGF-β pathway on the osteogenic differentiation effect of PTH

Received:2015-11-20 Revised:2015-12-19 Online:2016-03-25 Published:2016-04-06

摘要/Abstract

摘要： 目的：初步探究ClC-3氯通道在PTH调节成骨分化过程中的作用机制。方法：采用基因转染的方法，分别用特异性siRNA调节成骨细胞MC3T3-E1中转录生长因子TGF-β1及ClC-3氯通道基因的表达水平，采用Real-time PCR检测Clcn3、Tgfb1及成骨相关因子(Alp、Bsp、Oc 及Runx2)的基因表达情况，并用Western Blot及免疫荧光方法检测CLC-3氯通道蛋白及TGF-β1蛋白的表达情况。结果：在PTH间断刺激下，成骨细胞中仅干扰TGF-β1基因表达后，ClC-3氯通道基因及蛋白表达水平增强；仅干扰ClC-3氯通道基因后，TGF-β1基因及蛋白表达水平升高。同样在PTH间断刺激下，在分别干扰ClC-3氯通道基因和TGF-β1基因后，成骨相关基因(Alp、Bsp、Oc及Runx2)表达水平较对照组明显降低（P<0.05）。然而共同干扰两者之后成骨相关基因表达较对照组无明显差异（P>0.05）。结论：ClC-3氯通道通过TGF-β通路参与介导了PTH在成骨细胞中促进成骨细胞分化的作用，其具体的作用机制还需要更深入的研究。

Abstract: Objective：To investigate the mechanism of ClC-3 chloride channel in the progress of PTH regulating bone osteogenic differentiation. Methods：Regulate the expression of TGF-β1 and ClC-3 chloride channel in MC3T3-E1 cells with specific siRNA. Investigate the expression of Clcn3、Tgfb1 and osteogenic factors (Alp, Bsp, Oc and Runx2) with the method of Real-time PCR. Investigate the expression of CLC-3 chloride channel protein and TGF-β1 protein with the method of Western Blot and immunofluorescence. Results: The effect of TGF-β1 gene expression on ClC-3 expression in osteoblasts under discontinuous stimulation of PTH: ClC-3 chloride channel gene and protein expression increased when TGF-β1 gene expression in osteoblasts was interrupted only. The effect of ClC-3 chloride channel gene expression on TGF-β1 expression in osteoblasts under discontinuous stimulation of PTH: TGF-β1 gene and protein expression increased when ClC-3 chloride channel gene expression was interrupted only. Expression of osteogenic genes under discontinuous stimulation of PTH: Expression of osteogenic genes (Alp, Bsp, Oc and Runx2) decreased comparing to control when ClC-3 gene and TGF-β1 gene were interrupted respectively (P<0.05). However, no significant difference was observed between the expression of the simultaneously interrupted genes and the control （P>0.05）. Conclusion：ClC-3 chloride channel has been involved in the osteoinduction of PTH in the osteoblasts through the TGF-β pathway. The specific mechanism behind that requires further research.

中图分类号:

R783.5

张瑞王欢刘竹影沈鹏丁寅. ClC-3氯通道参与TGF-β通路调控PTH的促成骨分化作用机制研究[J]. , 2016, 7(1): 1-5.

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ClC-3氯通道参与TGF-β通路调控PTH的促成骨分化作用机制研究

ClC-3 chloride channel in the regulation of TGF-β pathway on the osteogenic differentiation effect of PTH

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