口腔生物医学
口腔生物醫學
구강생물의학
Oral Biomedicine
2015年
3期
137-142
,共6页
骨髓间充质干细胞%绝经后骨质疏松%miR-21%成骨分化%小鼠
骨髓間充質榦細胞%絕經後骨質疏鬆%miR-21%成骨分化%小鼠
골수간충질간세포%절경후골질소송%miR-21%성골분화%소서
Bone marrow mesenchymai stem ceiis%Postmenopausai osteoporosis%miR-21%Osteogenic differentiation%Mice
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.
目的:尋找雌激素缺乏所導緻的骨質疏鬆環境下調控小鼠骨髓基質榦細胞( mouse bone marrow stromai stem ceiis , mBMSCs )成骨的關鍵miRNA,併探討此miRNA在雌激素缺乏所導緻的骨質疏鬆微環境下是否參與調控mBMSCs成骨及其在此種微環境下對成骨的調控作用. 方法:建立卵巢切除動物模型,通過生物信息學技術以及miRNA基因芯片技術對卵巢切除組和假手術組的C57BL/6J小鼠來源的mBMSCs進行對比篩選,確定調控mBMSCs成骨的關鍵miRNA;利用實時定量RT-PCR技術驗證此miRNA在兩組mBMSCs成骨分化過程中的錶達差異;通過細胞轉染技術上調和下調此miRNA,實時定量RT-PCR、Western biot、茜素紅和堿性燐痠酶染色等技術觀察轉染後mBMSCs的成骨能力. 結果:生物信息學技術以及miRNA基因芯片技術篩選確定調控mBMSCs成骨的關鍵miRNA為miR-21;實時定量RT-PCR顯示miR-21在卵巢切除組mBMSCs成骨分化中的水平較假手術組低;轉染miR-21至卵巢切除組mBMSCs,能部分恢複其成骨分化能力. 結論:miR-21是雌激素缺乏所導緻的骨質疏鬆環境中調控mBMSCs成骨分化的關鍵miRNA;miR-21在雌激素缺乏所導緻的骨質疏鬆環境中能促進mBM-SCs的成骨分化.
목적:심조자격소결핍소도치적골질소송배경하조공소서골수기질간세포( mouse bone marrow stromai stem ceiis , mBMSCs )성골적관건miRNA,병탐토차miRNA재자격소결핍소도치적골질소송미배경하시부삼여조공mBMSCs성골급기재차충미배경하대성골적조공작용. 방법:건립란소절제동물모형,통과생물신식학기술이급miRNA기인심편기술대란소절제조화가수술조적C57BL/6J소서래원적mBMSCs진행대비사선,학정조공mBMSCs성골적관건miRNA;이용실시정량RT-PCR기술험증차miRNA재량조mBMSCs성골분화과정중적표체차이;통과세포전염기술상조화하조차miRNA,실시정량RT-PCR、Western biot、천소홍화감성린산매염색등기술관찰전염후mBMSCs적성골능력. 결과:생물신식학기술이급miRNA기인심편기술사선학정조공mBMSCs성골적관건miRNA위miR-21;실시정량RT-PCR현시miR-21재란소절제조mBMSCs성골분화중적수평교가수술조저;전염miR-21지란소절제조mBMSCs,능부분회복기성골분화능력. 결론:miR-21시자격소결핍소도치적골질소송배경중조공mBMSCs성골분화적관건miRNA;miR-21재자격소결핍소도치적골질소송배경중능촉진mBM-SCs적성골분화.
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 .