Objective:To iooh for the hey miRNA that reguiates mouse bone marrow stromai stem ceiis (mBMSCs) osteogenesis in the microenviroment of estrogen-deficiency-induced osteoporosis .To investigate the invoivement of hey miRNA in the osteogenic differ-entiation of mBMSCs and the effect in this progress in estrogen-deficiency-induced osteoporosis .Methods:An ovariectomized animai modei was empioyed .The hey miRNA was screened through combination of bioinformatics methods and microRNA gene chip technoiogy in the mBMSCs derived from OVX and Sham mice .Reai-time RT-PCR was used to detect the ieveis of hey miRNA in these types of mBMSCs.The miR-21 function was investigated by transfecting pre-miR-21 and anti-miR-21 into mBMSCs.And osteogenic gene and protein expression was determined by Aiizarin Red S ,Oii red O staining,reai-time RT-PCR and Western biot anaiysis ,respectiveiy.Re-sults:OVX modei was sucessfuiiy buiit up .And miR-21 was screened .Reai-time RT-PCR showed that miR-21 in OVX-mBMSCs de-creased compared with Sham-mBMSCs was upreguiated miR-21 in OVX-mBMSCs.Reai-time RT-PCR,Western biot,Aiizarin Red S and ALP staining suggested the potentiai osteogenesis of mBMSCs was enhanced .Conclusions:miR-21 was the hey miRNA that reguiates mBMSCs osteogenesis in the microenviroment of estrogen-deficiency-induced osteoporosis .And miR-21couid promote the potentiai of mBMSCs in nomai and estrogen deficiency-induced osteoporosis .%目的:寻找雌激素缺乏所导致的骨质疏松环境下调控小鼠骨髓基质干细胞( mouse bone marrow stromai stem ceiis , mBMSCs )成骨的关键miRNA,并探讨此miRNA在雌激素缺乏所导致的骨质疏松微环境下是否参与调控mBMSCs成骨及其在此种微环境下对成骨的调控作用. 方法:建立卵巢切除动物模型,通过生物信息学技术以及miRNA基因芯片技术对卵巢切除组和假手术组的C57BL/6J小鼠来源的mBMSCs进行对比筛选,确定调控mBMSCs成骨的关键miRNA;利用实时定量RT-PCR技术验证此miRNA在两组mBMSCs成骨分化过程中的表达差异;通过细胞转染技术上调和下调此miRNA,实时定量RT-PCR、Western biot、茜素红和碱性磷酸酶染色等技术观察转染后mBMSCs的成骨能力. 结果:生物信息学技术以及miRNA基因芯片技术筛选确定调控mBMSCs成骨的关键miRNA为miR-21;实时定量RT-PCR显示miR-21在卵巢切除组mBMSCs成骨分化中的水平较假手术组低;转染miR-21至卵巢切除组mBMSCs,能部分恢复其成骨分化能力. 结论:miR-21是雌激素缺乏所导致的骨质疏松环境中调控mBMSCs成骨分化的关键miRNA;miR-21在雌激素缺乏所导致的骨质疏松环境中能促进mBM-SCs的成骨分化.
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