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目的探讨雌激素(Estrogen)缺乏对骨髓间充质干细胞(BMMSCs)表达Fas水平及受破骨细胞凋亡的影响。方法建立绝经后骨质疏松模型(切除双侧卵巢,去势组,OVX),选用同一批次周龄相同、体质量相近的健康小鼠行双侧卵巢附近脂肪组织部分切除,建立假手术组(sham)。将破骨细胞与2组BMMSC共培养,Annexin V/7-AAD/Sca-1流式细胞仪计数检测2组细胞中BMMSC凋亡的差异;Western blot检测共培养组中Bcl-2、cleaved-caspase-3蛋白表达情况;RT-PCR和免疫荧光检测sham组、OVX组中Fas表达差异。结果 OVX组中BMMSC的凋亡程度较sham组显著增强(P<0.05)。Western blot显示OVX共培养组中Bcl-2、cleaved-caspase-3表达强度较sham共培养组显著上调(P<0.05)。免疫荧光染色实验显示OVX组Fas表达强度较sham组显著增强(P<0.05)。结论雌激素调控BMMSCs中Fas的表达,并且雌激素影响破骨细胞对BMMSC的凋亡,这可能是绝经后骨质疏松发病的重要因素之一。
Objective To investigate the effect of estrogen deficiency on the expression of Fas in bone marrow mesenchymal stem cells (BMMSCs) and the apoptosis of osteoclasts. Methods The model of postmenopausal osteoporosis (ovariectomized ovariectomized, ovariectomized, ovariectomized, ovariectomized, ovariectomized, ovariectomized, ovariectomized, ovariectomized, ovariectomized) was established. (sham). The osteoclasts were co-cultured with BMMSCs from two groups. The difference of apoptosis of BMMSCs in two groups was detected by Annexin V / 7-AAD / Sca-1 flow cytometry. Western blot was used to detect the expression of Bcl-2 and cleaved- caspase-3 protein expression; RT-PCR and immunofluorescence sham group, OVX group Fas expression differences. Results The apoptosis of BMMSCs in OVX group was significantly higher than that in sham group (P <0.05). Western blot showed that the expression of Bcl-2 and cleaved-caspase-3 in OVX co-culture group was significantly up-regulated compared with sham co-culture group (P <0.05). Immunofluorescence staining showed that the expression of Fas in OVX group was significantly higher than that in sham group (P <0.05). Conclusion Estrogen regulates the expression of Fas in BMMSCs, and estrogen affects the apoptosis of BMMSCs induced by osteoclasts, which may be one of the important factors in the pathogenesis of postmenopausal osteoporosis.