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目的:检测雌激素浓度与小鼠骨髓间充质干细胞(mBMMSCs)凋亡的相关性,初步探讨雌激素调节mBMMSCs凋亡的分子机制。方法:构建雌性C57BL小鼠去势模型,模拟绝经期妇女低雌激素体质。分别取去势和假手术组小鼠以及正常小鼠mBMMSCs进行原代分离、培养和鉴定。利用流式细胞术(FCM)比较去势和假手术两组细胞的凋亡差异,以及不同雌激素浓度培养条件对正常C57BL小鼠mBMMSCs凋亡的影响。参考文献中芯片结果,利用实时定量PCR方法检测miR-21在去势组及假手术组mBMMSCs中的表达差异,以及不同雌激素浓度培养条件下正常小鼠mBMMSCs中miR-21的表达差异。结果:成功构建了去势小鼠模型。利用FCM证实去势小鼠骨髓间充质干细胞在体外培养的过程中较假手术组更易凋亡。正常小鼠mBMMSCs体外培养过程中,随着雌激素浓度的升高,其凋亡减少。去势鼠来源mBMMSCs中miR-21表达量低于假手术组来源mBMMSCs。高浓度雌激素培养液培养的mBMMSCs中miR-21表达高于低浓度雌激素培养液培养的mBMMSCs。结论:雌激素对mBMMSCs的凋亡具有抑制作用,miR-21可能参与这一过程。
Objective: To detect the correlation between estrogen concentration and the apoptosis of mouse bone marrow mesenchymal stem cells (mBMMSCs), and to explore the molecular mechanism of estrogen regulating the apoptosis of mBMMSCs. METHODS: Female C57BL mice castration model was constructed to simulate low estrogen in postmenopausal women. Mice were removed and sham-operated mice and normal mice mBMMSCs for primary isolation, culture and identification. Flow cytometry (FCM) was used to compare the apoptosis of castrated and sham-operated cells, and the effect of different estrogen concentrations on the apoptosis of mBMMSCs in normal C57BL mice. References Chips results, the use of real-time quantitative PCR detection of miR-21 in castration group and sham operation group mBMMSCs differences in expression, and different estrogen concentrations in normal mouse mBMMSCs miR-21 expression differences. Results: The castrated mouse model was successfully constructed. FCM confirmed that castrated mouse bone marrow mesenchymal stem cells were more susceptible to apoptosis than the sham-operated group during in vitro culture. In normal mouse mBMMSCs cultured in vitro, with the estrogen concentration increased, the apoptosis decreased. The expression of miR-21 in mBMMSCs derived from the castration mice was lower than that of mBMMSCs from the sham-operated group. The expression of miR-21 in mBMMSCs cultured in high concentration of estrogen was higher than that in mBMMSCs cultured in low concentration of estrogen. Conclusion: Estrogen can inhibit the apoptosis of mBMMSCs, and miR-21 may be involved in this process.