Effect Of Rosuvastin On The Proliferation And LRP5and Runx2Gene Expression In Vitro Cultured Human Osteoblasts

ObjectiveObserved with different concentration Rosuvastatin after24h on cultured human bone cells value-added and low-density lipoprotein receptor-relatedprotein5(LRP5)、Runx2gene expression in. Preliminary study of rosuvastatin affect the possible molecular mechanisms of bone metabolism.Methods1. Collection of adult males iliac cancellous bone (Second Clinical Medical College of Shanxi Medical orthopedic operating room in normal surgical procedure). The use of isolated and purified enzyme digestion osteoblasts in vitro cultured human osteoblasts, to observe the morphology and function.2. Human osteoblasts in vitro experimental models, With different concentr ations (010-910-810-710-6mol/l) of rosuvastatin stimulate the logarithmic phase of growth osteoblasts for24hours, cell proliferation ability was detected with ckk-8coiorimetric assay and then determine their OD mv.3. In osteoblasts passage process, take5bottles of good growth, the growth rate of70-80%of osteoblasts which are culture with different dose of rosuvastatin (010-910-810-710-6mol/l) for24hours, discard the culture medium, Were extracted using Trizol method total RNA, Using fluorescence quantitative PCR assay for detection of LRP5, Runx2in the relative expression levels4. Statistical analysis:data are presented as x±s. All statistical analysis is performed with windows SPSS17.0. The different between the group are determined by One-way ANOVE. The different between RNAdom two groups are determined by LSD. Differences are considered significant at a value of P<0.05.Results1.Cultured human osteoblasts inverted microscope, Adherent primary cell cultures3d, fusiform、polygonal common,12-15d monolayer covered sidewall. Passage cells are mostly spindle-shaped or triangular, large nuclei. Osteocalcin immunohistochemistry staining cells positive reaction, the cytoplasm visible brown gRNAular substance. And the calcium alizarin red staining was positive.2. With different concentrations (010-910-810-710-6mol/l) of rosuvastatin stimulate the in vitro culture of human osteoblasts after24hours, Using observed under the microscope of CCK-8colorimetric, Significantly increased the number of osteoblasts, each OD are increased,10-7M is the most obvious value.The experimental groups and compared with the control groups were considered significant at a value of P<0.01.3. With different concentrations (010-910-810-710-6mol/1) of rosuvastatin stimulate the in vitro culture of human osteoblasts after24hours, Using fluorescence quantitative PCR assay for detection of LRP5, Runx2in the relative expression levels. The results show:rosuvastatin promote LRP5、Runx2mRNA expression,10-7M is the most obvious value. The experimental groups and compared with the control groups were considered significant at a value of P<0.01.Conclusions1. Rosuvastin promotes osteoblast proliferation2. Rosuvastin promotes expression of LRP5、Runx2,so as to osteoblasts differentiation,accordingly promote bone formation.