Expression Of Runx2 And PPAR-γ In Bone Marrow Mesenchymal Stem Cells Of Ovariectomized Rats

Objective To investigate the expression of Runx2 and PPAR-γ in bone marrow mesenchymal stem cells of estrogen, puerarin and reseveratrol treated ovariectomized rats.Methods 1. Sixty-five healthy three-month-old female Sprague-Dawley rats were randomly divided into five groups, i.e sham-operated group(SHAM), ovariectomized group(OVX), ovx+estrogen replacement therapy(ERT), ovx+puerarin(PUE) and ovx+resveratrol(RES). Each group includes thirteen rats. All the ovariectomized rats were the osteoporosis model except the SHAM group. Intervening treatment last 10 weeks since we started 1 week after the surgery. 2. Using the ELISA kits to measure the quantity of bone specific alkaline phosphatase(BALP), osteoproprotegrin(OPG), tartrate-resistant acid phosphtase-5b(TRAP-5b) and Cross-linked C-telopeptides of type I collagen(CTX-I). Taking the peripheral quantitative computed tomography(pQCT) to detect relevance parameters of right femur’s bone microstructure. 3. Bone marrow mesenchymal stem cells from rats were isolated by whole bone marrow adherence method. The cell surface antigens(CD90/CD45) of bone marrow mesenchymal stem cells were detected by flow cytometry(FCM). Then we examined the Runx2, BMP-2,PPAR-γ and LPL mRNA expression level of the bone marrow mesenchymal stem cells by RT-PCR.Results 1. Compared with the SHAM, the weight of rats in OVX increased by 3.9% and8.4% respectively after surgery taken 4 and 14 weeks happened in the ERT, PUE and RES groups had the increasing trend. 2. By comparing with the sham group, the cancellous bone mineral and bone density in the OVX group decreased(P<0.05). But they were increased in the PUE, RES and ERT group. Meantime. There was no significance of mineral content and density of cancellous bone in the PUE, RES, ERT and OVX group. 3. Compared with SHAM, the serum level BALP,CTX-I and TRAP-5b were increased. The serum level of those markers were decreased in ERT,PUR, RES group.Compared with SHAM, the level of OPG in OVX group was decreased 36.8%(P<0.01) after 11 weeks. 4. The isolated BMSCs without any intervention covered the bottom of the bottle on nineteenth days by inverted microscope. BMSCs were negative for CD45 and positive for CD90 by FCM. 5. Examining the expression level of Runx2, BMP-2, PPAR-γ andLPL mRNA in the bone marrow mesenchymal stem cells by RT-PCR, but there was no significance in each group except that the PPAR-γ expression was lower in ERT group.Conclusion There was no significant difference in Runx2, BMP-2, PPAR-γ and LPL mRNA expression in BMSCs of estrogen, puerarin and reseveratrol treated ovariectomized rats in vitro. It was probable that BMSCs might keep the balance of cell differentiation via self regulation.It might brought a new idea in osteoporosis treatment via intervention of auto-bone marrow mesenchymal stem cell.