孕激素对成骨细胞凋亡的影响及其机制  

Effect and mechanism of progesterone on apoptosis of osteoblastic cells

作　　者：王清平[1] 谢辉[1] 唐四元[2] 王旦[1] 李辉[1] 罗湘杭[1] 廖二元[1] 

机构地区：[1]中南大学湘雅二医院代谢内分泌研究所,长沙410011 [2]中南大学护理学院

出　　处：《中华老年医学杂志》2010年第4期324-327,共4页Chinese Journal of Geriatrics

基　　金：国家自然科学基金（30872708,30600661）;2008年湖南省博士后科研专项计划（2008RS4012）

摘　　要：目的探讨孕激素对成骨细胞凋亡的影响及其机制。方法以Western blot分析测定鼠原成骨细胞（MC3T3-E1）孕激素受体的表达水平和细胞色素C释放，吖啶橙／溴乙碇（AO／EB）染色和夹心法酶免法分析测定细胞凋亡，荧光底物显色法估计半胱天冬蛋白酶-9／3的活性。结果在孕激素浓度为10^-10mol／L、10^-9mol／L及10^-8mol／L细胞培养中，细胞凋亡程度分别为1．93±0．12、1．57±0．13和0．79±0．14，均明显低于无血清细胞培养（2．97±0．22），差异有统计学意义（均为P〈0．01）；添加10μmol／L孕激素受体拮抗剂RU486后，细胞凋亡程度与无血清培养差异无统计学意义（2．88±0．21，P〉0．05）；孕激素对细胞色素C的释放及半胱天冬蛋白酶-9／3活性的抑制作用均被逆转。结论孕激素通过激素受体及其下游的线粒体信号通路抑制成骨细胞凋亡，这可能与孕激素促进骨形成的作用机制有关。Objective To investigate the effect and mechanism of progesterone on apoptosis of murine MC3T3-E1 osteoblastic cells. Methods The cell apoptosis was measured by acridine orange/ethidium bromide （AO/EB） staining and sandwich-enzyme-immunoassay. Progesterone receptor （PR）, cytochrome c, caspase-9 and caspase 3 protein levels were determined by Western blot analysis. The enzyme substrate was also used to assess the activation of caspase-3 and caspase-9. Results After 24 h of starvation, apoptotic cells at progesterone of 10^-10 mmol/L （1.93 ± 0. 12 ELISA absorbance units）, 10^-9 mmol/L （1.57±0.13 ELISA absorbance units） and 10^-8 mmol/L （0.79±0.14 ELISA absorbance units） were less than in vehicle-treated group （2.97±0.22 ELISA absorbanee units, all P〈0.05）, but this effect was blocked by a PR antagonist RU486. Furthermore, the suppressive effects of progesterone on cytochrome c release and caspase-9, easpase-3 activation in serum-deprived MC3T3-E1 cells were also reversed by RU486. Conclusions The progesterone protects osteoblast against apoptosis through PR and the downstream mitochondrial pathway. Thus, the data suggest that the effects of progesterone on osteoblast apoptosis may contribute to the mechanisms by which progesterone exerts its action on bone formation.

关 键 词：孕激素 成骨细胞 细胞凋亡 

分 类 号：R335[医药卫生—人体生理学]