雌激素受体α36参与淫羊藿素对MG63细胞的促增殖和抗凋亡作用
刘伟¹, 罗自强², 任璐¹, 孙玫¹, 姚菊琴¹, 王井霞¹, 王尊¹, 唐四元^1,*
1中南大学湘雅护理学院，长沙 410000；²中南大学基础医学院，长沙 410000
摘要
淫羊藿素能够显著改善绝经后骨质疏松(postmenopausal osteoporosis, PMOP)，其作用机制尚未阐明。本研究旨在探讨雌激素受体α36 (estrogen receptor α36, ERα36)在淫羊藿素对成骨细胞促增殖和抗凋亡中的作用及其下游信号转导机制。通过转染shRNA载体构建ERα36敲减的人成骨肉瘤MG63细胞模型，用CCK-8检测细胞增殖情况，用流式细胞术检测细胞凋亡，用Western blot检测细胞内ERK和AKT信号通路的激活情况。结果显示，ERα36敲减后，淫羊藿素对MG63细胞的促增殖和抗凋亡作用显著减弱；淫羊藿素可上调MG63细胞内ERK及AKT磷酸化水平，该作用可被ERα36敲减取消；U0126 (ERK信号通路阻断剂)和LY294002 (AKT信号通路阻断剂)分别预处理后，淫羊藿素对MG63细胞的促增殖作用均显著减弱；U0126预处理后，淫羊藿素对MG63细胞的抗凋亡作用显著减弱；而给予LY294002预处理后，淫羊藿素对MG63细胞的抗凋亡作用无明显变化。以上结果提示，淫羊藿素通过ERα36及其下游的ERK及AKT信号通路发挥了对成骨细胞的促增殖和抗凋亡作用。


关键词： 骨质疏松; 雌激素受体α36; 淫羊藿素; 成骨细胞
分类号：R589

ERα36 is involved in the proliferation promotion and anti-apoptosis effects of icaritin on MG63 cells
Liu Wei¹, LUO Zi-Qiang², REN Lu¹, SUN Mei¹, YAO Ju-Qin¹, WANG Jing-Xia¹, WANG Zun¹, TANG Si-Yuan^1,*
1Xiangya Nursing School, Central South University, Changsha 410000, China；²School of Basic Medical Science, Central South University, Changsha 410000, China
Abstract
The mechanism for icaritin to improve postmenopausal osteoporosis (PMOP) has not been clarified. The aim of this study was to investigate the role of estrogen receptor α36 (ERα36) in the proliferation promotion and anti-apoptosis effects of icaritin on osteoblasts and the underlying mechanism of downstream signal transduction. The ERα36 knockdown human osteosarcoma MG63 cell model was constructed by transfection of shRNA vector. Cell proliferation was detected by CCK-8, the apoptosis was detected by flow cytometry, and the activation of ERK and AKT signaling pathways was detected by Western blot. The results showed that the effects of icaritin on the proliferation and apoptosis of MG63 cells were significantly decreased after ERα36 knockdown, and icaritin could up-regulate the levels of ERK and AKT phosphorylation in MG63 cells, which could be reduced by ERα36 knockdown. The effect of icaritin on the proliferation of MG63 cells was significantly decreased by pretreating the cells with U0126 (an ERK signaling pathway blocker) and LY294002 (an AKT signaling pathway blocker), respectively. Furthermore, anti-apoptotic effect of icaritin on MG63 cells was significantly decreased after the cells were pretreated with U0126, but not with LY294002. These results suggest that icaritin exerts proliferation promotion and anti-apoptosis effects on osteoblasts through ERα36 and its downstream ERK and AKTsignaling pathways.
Key words: osteoporosis; estrogen receptor α36; icaritin; osteoblast

收稿日期：2018-05-12　　录用日期：2018-09-26
通讯作者：唐四元　　E-mail: sytang263@outlook.com
DOI: 10.13294/j.aps.2018.0065
引用本文：
刘伟, 罗自强, 任璐, 孙玫, 姚菊琴, 王井霞, 王尊, 唐四元. 雌激素受体α36参与淫羊藿素对MG63细胞的促增殖和抗凋亡作用[J]. 生理学报 2018; 70 (5): 474-480.
Liu Wei, LUO Zi-Qiang, REN Lu, SUN Mei, YAO Ju-Qin, WANG Jing-Xia, WANG Zun, TANG Si-Yuan. ERα36 is involved in the proliferation promotion and anti-apoptosis effects of icaritin on MG63 cells. Acta Physiol Sin 2018; 70 (5): 474-480 (in Chinese with English abstract).



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