The Effection Of Imprinted Genes (H19,Grb10) And MiR-675 On The Proliferation And Differentiation In Bone Healing

Objective : Imprinted genes are very important for fetal growth, cell metabolism,differentiation of stem cells. Bone injury healing is a process of local bone development,it is an extremely complex biological process involved variety of factors.Research on the relationship between genes and micro RNA imprinting and bone damage repair is small. The present study was to detect H19, miR-675, Grb10 expression in bone healing process explore the relationship between bone healing cells and the imprinted genes and micro RNA.Methods: Left hind femur bone defect modeling, the right hind femur sham operation, 5d,10 d, 15 d, 20 d, 25 d kill them by surgery decapitated. And their femoral bones were removed.After conventional fixed, embedded, slices. Using Western blot and immunohistochemistry to observe the expression of Grb10 protein. And using real-time quantitative PCR to detect the expression of H19?miR-675.Results: Immunohistochemical staining found that: The expression of Grb10 protein in the5 d and 10 d experimental group are increasing, the average optical density values ??showed that the protein expression of experimental group 5d and 10 d is twice the control group.Real-time PCR results showed that: H19, Grb10, miR-675 was expressed in 5d to 25 d injury group and control group; after injury 5d, the experimental group H19 expression is twice the control group, after injury 10 d, H19 expression in the experimental group is also twice the control group; expression after injury 15 d, 20 d, 25 d in the experimental group and the control group was essentially flat; all the control group express miR-675 consistently. 5d, The expression of miR-675 in the experimental group after injury volume was 2.5 times the control group, after injury 10 d, The expression of miR-675 in experimental group increased three times in the control group; 15 d, 20 d, 25 d, the experimental and control groups was essentially flat; after injury 5d in the experimental group, the Grb10 expression level are three times the control group, after injury 10 d the experimental group was decreased, compared with the control group, the control group three times. Western blot results showed that: After injury 5d Grb10 protein is the control group twice, in the 10 d experimental group decreased protein expression relative to the control group also increased by about two times, 15 d, 20 d,25d experiment basically the same group and the control group protein content.Conclusion:1) H19, Grb10 and miR-675 to promote the proliferation and differentiation of osteoblasts, accelerate wound healing process.