中文摘要：

目的：探讨并验证重组miRNA-30a-5p体外转染对人骨髓间充质干细胞（MSCs）向成骨细胞分化的生物学作用。方法：人工重组合成从人骨髓MSCs定向成骨分化差异性表达miRNA基因芯片结果中筛选成骨性表达量显著增高的miRNA-30a-5p。从人体骨髓中分离培养MSCs,成骨诱导分化过程中于体外转染重组miRNA-30a-5p,并通过茜素红S法染色检测钙盐沉积、碱性磷酸酶（ALP）钙钴法染色及ALP比色法定量检测成骨细胞ALP活性,对比观察研究重组miRNA-30a-5p在人骨髓MSCs定向成骨诱导分化过程中的生物学功能。结果：成功分离培养人骨髓MSCs并诱导其成骨定向分化。经体外向MSCs转染miRNA-30a-5p并诱导其成骨定向分化,转染效率为（37.32±2.43）%。分别于诱导培养第14、21天行ALP染色观察、ALP活性测定、茜素红钙盐结节染色检测各组细胞成骨活性,结果显示miRNA-30a-5p mimics转染组MSCs成骨分化特性显著性增强（P〈0.05）。结论：miRNA-30a-5p在MSCs定向成骨分化的过程中具有一定的促进增强作用,被转染的MSCs向成骨细胞定向分化表达有所增加,为进一步阐明人骨髓MSCs定向成骨分化的分子生化机制,细胞移植修复治疗骨缺损奠定了理论基础。

英文摘要：

Objective： To investigate and verify biological effects of the human bone marrow mesenchymal stem cells（MSCs） in vitro transfection with recombinant miRNA-30a-5p in differentiation into osteoblasts.Methods： We selected miRNA-30a-5p with significantly hight expression of osteoblast through osteogenic differentiation and miRNA differentially expressed genes microarray from hBMSC,and then synthesize the recombinant human miRNA-30a-5p.Separated and identified human bone marrow MSCs and induced them into osteoblasts through in vitro transfection of recombinant miRNA-30a-5p.Detected the activity of ALP in osteoblast by alkaline phosphatase（ALP） staining and ALP calcium cobalt quantitative detection,calcium deposition by alizarin red S staining.And investigate the biological functions of the recombinant miRNA-30a-5p in the differentiation of the human mesenchymal stem cells into osteoblasts in comparative observation.Results： Successfully we isolated and cultured MSCs from human bone marrow and inducted the into bone through directional differentiation,The transfection efficiency was（37.32±2.43）%.At different time points（day 14,day 21） after induction,osteogenetic activity of the cells was detected by ALP staining observation,ALP activity detection and alizarin red staining for calcium nodules,the results showed that transfection of artificial re-synthesis miRNA-30a-5p into stem cells significantly enhanced features of stem cells differentiating into bone.Conclusion： miRNA-30a-5p has a certain role in promoting orientational differentiation of the MSCs into bone cells.MSCs directely differentiation into osteoblasts is increased by transfected with the recombinant miRNA-30a-5p.That has fundamentally value in elucidating molecular and biochemical mechanisms of osteogenic differentiation and establishing the theoretical basis for cell transplantation of bone defect repair treatment.