中国血液净化

中国血液净化 ›› 2014, Vol. 13 ›› Issue (09): 647-649.doi: 10.3969/j.issn.1671-4091.2014.09.010

• 基础研究 • 上一篇    下一篇

甲状旁腺激素对HAEC 细胞EndMT 诱导作用及机制探讨

李宁,李亚斐   

  1. 第三军医大学军事预防医学院(李宁现在第三军医大学第一附属医院肾内科工作）
  • 收稿日期:2014-03-31 修回日期:2014-06-19 出版日期:2014-09-12 发布日期:2014-09-02
  • 通讯作者: 李亚斐 liyafei2008@hotmail.com E-mail:liyafei2008@hotmail.com

The effect and mechanism of parathyroid hormone induced endothelial-mesenchymal transition of vascular endothelial HAEC cells

  • Received:2014-03-31 Revised:2014-06-19 Online:2014-09-12 Published:2014-09-02

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摘要： 目的探讨甲状旁腺激素(parathyroid hormone,PTH)是否诱导血管内皮细胞上皮细胞-间充质转化(endothelial-mesenchymal transition, EndMT)及相关机制。方法血管内皮细胞HAEC 培养至对数期，分别加入不同剂量的全长重组PTH(10-12 mol/L、10-11 mol/L、10-10mol/L、10-9mol/L、10-8 mol/L)作用48h；再在10-8 mol/L PTH 作用下，选择12、24、36、60h 4 个时间点；分别收集总蛋白。Western blot分析Ve-Cadherin、CD31、α-SMA、TGF-β1 和ILK 的表达改变。结果Western blot 结果表明在PTH 作用下，血管内皮细胞标志分子Ve-Cadherin 和CD31 表达降低；纤维细胞标志分子α-SMA 表达则显著升高，且呈时间和剂量依赖性。Western blot 分析发现TGF-β1 和ILK 表达同时升高。结论PTH 减弱内皮细胞特征、增强纤维化特征，诱导HAEC 细胞发生EndMT，TGF-β1-ILK 通路是其调控的可能通路之一。

关键词: 甲状旁腺素, 内皮细胞-间充质转化, 尿毒症, 整合素连接激酶

Abstract: Objective To investigate whether the increased parathyroid hormone (PTH) as a uremic toxin induces the endothelial-mesenchymal transition (EndMT) of vascular endothelial cells (HACE cells) and its related mechanisms. Methods The vascular endothelial HACE cells were cultured to logarithmic phase, and
PTH at the concentration of 10-12, 10-11, 10-10, 10- 9, and 10-8 mol/L was added into the medium for 48 h. Additionally, HACE cells were cultured in the medium containing 10-8 mol/L PTH for 12, 24, 36, and 60 h. Cells were then harvested for morphological examination and for VECadherin, CD31, α-SMA, TGF-β1 and ILK expression determinations by western blot. Results Morphological examination showed that the HACE cells tended to have fibrosis changes after the PTH induction. Western blot displayed that PTH reduced the expression of vascular endothelial cell markers CD31 and VECadherin and increased the expression of fiber cell markers α-SMA in timE and dosEdependent manners. At the same time the expressions of TGF-β1 and ILK increased. Conclusion PTH induced the HAEC cells to have less endothelial cell characteristics and more fibrosis characteristics, indicating the presence of EndMT in HACE cells. The TGF-β1-ILK pathway may be involved in the PTH-induced changes.

Key words: Parathyroid hormone, Endothelial- mesenchymal transition (EndMT), Uremia, Integrinlinked kinase