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Objective:To investigate the effect of miR-204 on the proliferation and differentiation of osteoblasts in osteoporosis mice by Wnt signaling pathway and its mechanism.Methods:Female Kunming mice were divided into: control group, sham operation group and osteoporosis group. Ovariectomy mouse models were established and identified by bilateral ovariectomy; Mouse primary osteoblasts were extracted and identified; Cells was transfected and detected the miR-204 expression levels; MTT was used to detect the viability of each group of cells; Alkaline phosphatase (ALP) activity was detected in each cell group; Cell flowmetry was used to detect apoptosis in each group; Cell flowmetry was used to detect the activity of Caspase-3 in each group of cells; Interaction between miR-204, β-catenin and LRP-5 was detected by dual luciferase reporter gene. Western blot was used to detect the expression of Wnt signaling pathway-related proteins.Results:The bone mineral density of the osteoporosis group was significantly lower than that of the control group and the sham operation group ( P=0.007, P=0.057) , indicating that the osteoporosis mice were successfully modeled; The expression level of miR-204 was significantly increased in the miR-204 mimics group ( P=0.007) , and decreased in the miR-204 inhibitor group ( P=0.031) ; The activity of bone cell and ALP activity of miR-204 mimics increased ( P=0.007, P=0.043) , and the activity of bone cell and ALP decreased by miR-204 inhibitor ( P=0.007, P=0.035) ; The invasive ability of miR-204 mimics was significantly increased ( P=0.006) , and the invasive ability of miR-204 inhibitor was decreased ( P=0.036) ; The apoptosis ability and Caspase-3 activity of miR-204 mimics were decreased ( P=0.041, P=0.045) , and the apoptosis ability and Caspase-3 activity of bone cells were enhanced by miR-204 inhibitor ( P=0.005, P=0.039) ; There were targeting relationship between miR-204 and β-catenin, LRP-5. The expressions of β-catenin and LRP-5 protein in osteoblasts of miR-204 mimics were up-regulated ( P=0.043, P=0.009) , and the expression of β-catenin and LRP-5 protein in bone cells of miR-204 inhibitor was down-regulated ( P=0.041, P=0.032) . Conclusion:miR-204 maybe promote the proliferation and differentiation of osteoblasts, activate Wnt signaling pathway, and has certain protective effect on osteoporosis.
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From the perspective of clinical doctors, in the teaching idea, national policy, school policies and different stages of clinicians cultivation , the article introduced the concept of translational medicine in the process of the clinical doctors tralning in the United States. It described US medical schools' (U.S. Virginia University School of medicine, for example) implementation ap-proach of translational medicine ideas in different stages, such as before entering school, during the period of school, practice exam and physician clinician tralning. It provided the reference for the de-velopment of translational medicine education in the process of China clinician education and tralning, including:medical students' integration into the related clinical research in preschool through volunteer service, the choice of multiple combination model of clinical science and basic research, interdisci-plinary examination of medical practitioners and the provisions of the research work in resident and specialist tralning stage.
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BACKGROUND:Pueraria is a member of isoflavone characterizing by decreasing the toxic effects of alcohol,relieving the effect of alcohol withdrawal,anti-oxidative effect,removing oxygen free radicals,and preventing cell injury induced by lipid peroxidation.OBJECTIVE:To investigate the inhibitory effect of puerarin on adipogenic differentiation of alcohol-induced human bone marrow mesenchymal stem cells (hBMSCs).DESIGN,TIME AND SETTING:An in vitro observation based on cytology was performed at Laboratory of Biochemistry and Molecular Biology,Basic Medical College of Zhengzhou University in October 2008.MATERIALS:The bone marrow was taken from the adult healthy volunteers from the Department of Orthopedics of the First Affiliated Hospital of Zhengzhou University.Puerarin standard was provided by the National Institute for the Control of Pharmaceutical and Biological Products.METHODS:The primary hBMSCs were purified from the adult marrow by gradient centrifugation.The second passaged cells were divided into 3 groups.Blank control group:Cells were cultured with DMEM culture media containing 10% fetal bovine serum;Model group:Cells were cultured with 0.09 mol/L alcohol;Experimental group:Cells were cultured with 0.09 mol/L alcohol and 10-6 mol/L puerarin;while,the culture liquid was changed every two or three days.Cells were cultured for 7 days in each group.MAIN OUTCOME MEASURES:PPARγ 2 gene expression with RT-PCR;amount of adipocyte with oil red "O" staining;alkaline phosphatase activity.RESULTS:PPAR-γ2 mRNA expression in the model group was significantly greater than experimental group and blank control group (P ＜0.05),while the expression in the experimental group was slightly greater than blank control group,but there was no significant difference (P ＞0.05).The number of adipocytes was the most in the model group,and the lipid droplet was large and abundant.The number of adipocytes in the experimental group was significantly less than model group (P ＜ 0.05),and there were few adipocytes in the blank control group.Compared with blank control group,alkaline phosphatase activity was decreased in the model group (P＜0.05);but the activity in the experimental group was increased compared to model group (P ＜ 0.05).CONCLUSION:Puerarin inhibits differentiation of hBMSCs into adipocytes induced by alcohol.