The Role Of MiR-483-5P In Osteogenic Differentiation Of Rat Bone Marrow Mesenchymal Stem Cells

Background:With the progress of social aging,the incidence of osteoporosis is also gradually increasing.Complications such as pain and fractures caused by osteoporosis have also become problems.The decrease of osteogenic differentiation ability of bone marrow mesenchymal stem cells(BMSCs)is considered to be one of the reasons for the onset and progression of osteoporosis.Therefore,it is of great significance to study the regulatory mechanism in the process of bone marrow mesenchymal stem cell differentiation and explore the methods to enhance the ability of bone marrow mesenchymal stem cell differentiation in the treatment of osteoporosis and other diseases.As a type of non-coding RNA,microRNA(miRNA)has received more and more attention on the regulation of cell functions.As people continue to study miRNAs,they gradually reveal the role of different miRNAs in cell differentiation,proliferation,apoptosis,migration and so on.However,the regulation of osteogenic differentiation of bone marrow mesenchymal stem cells by miR-483-5p has not been clearly studied.Objective:1.To study the changes of miR-483-5p during the osteogenic differentiation of bone marrow mesenchymal stem cells;2.To study the regulatory effect of miR-483-5p on the osteogenic differentiation of bone marrow mesenchymal stem cells.Methods:1.Extract bone marrow mesenchymal stem cells by whole bone marrow culture method from rats;detect the molecular expression on the cell surface by cell flow cytometry;identify the potential of osteogenic differentiation after inducing osteogenic differentiation by alkaline phosphatase staining and alizarin red staining;identify the adipogenic differentiation potential of cells by oil red O staining.2.Use RT-qPCR to test changes in miR-483-5p expression of bone marrow mesenchymal stem cells after osteogenic differentiation.3.Use lentivirus to construct different groups of bone marrow mesenchymal stem cells that overexpress or inhibit miR-483-5p;test the changes of osteogenic differentiation ability by alkaline phosphatase staining,alizarin red S staining,RT-qPCR,and Western-blot.Result:1.The rat bone marrow mesenchymal stem cells were successfully isolated and cultured.The mesenchymal stem cell-specific cell surface molecules were positively expressed,and the hematopoietic stem cell surface molecules were negatively expressed.Each staining result showed that BMSCs have the potentials of osteogenic and adipogenic differentiation.2.MiR-483-5p was significantly increased in BMSCs which were induced to osteogenic differentiation for 7 days;3.After overexpression of miR-483-5p,bone marrow mesenchymal stem cells enhanced osteogenic differentiation and inhibited miR-483-After 5p,the osteogenic differentiation of bone marrow mesenchymal stem cells decreased.Conclusion:1.The cells isolated in this experiment are rat bone marrow mesenchymal stem cells.2.MiR-483-5p can regulate the osteogenic differentiation of rat bone marrow mesenchymal stem cells;the overexpression of miR-483-5p can promote the osteogenic differentiation of BMSCs,while the inhibition of miR-483-5p can reduce the osteogenic differentiation of BMSCs.