Effect Of Parathyroid Interacts With Simvastatin On The Differentiation And The Gene Expression Of OPG、RANKL In Cultured Rat Osteoblast

Objective:To study the effect of recombinant human parathyroid hormone1-34(rhPTH1.34) interacts with Simvastatin on the differentiation and the gene expression of OPG、RANKL in cultured rat osteoblast。Investigate the possible mechanism of affect on osteoblasts, for clinical to provide futher theoretical basis。Methods:1. Culture suckling rat osteoblasts, and observe their morphology and function..2. The suckling rat osteoblasts are culture in vitro, the effect of different dose of rhPTH1-34(10-9mol/L) and Simvastatin (10-8、10-7、10-6、mol/L) on primary culture suckling rat osteoblasts differentiation is assessed by PNPP colorimetry after24hours, and then determine their OD mv.3. The suckling rat osteoblasts are culture with different dose of rhPTHi.34(10-9mol/L) and Simvastatin (10-8、10-7、10-6、mol/L) on primary culture suckling rat osteoblasts.The expression of OPG、RANKL mRNA in primary culture suckling rat osteoblasts is analyzed by semiquantative RT-PCR.4.Statistical analysis:data are presented as,x±s.All statistical analysis is performed with windows SPSS17.0.One-way ANOVE can determine the different between the group.LSD determine the different between random two groups. Differences are considered significant at a value of P<0.05.Results:l.Most of primary culture suckling rat osteoblasts were adhered of dish,and elongated spindle-shaped after3d,the cells are paved the layer of dish after 12-15d.ALP staining was positive,the ratio is about50-90%.The characteristics accord with the osteoblast features.2.Either intermittent rhPTH1-34(10-9mol/L) or different dose of SIM (10-8、10-7、10-6mol/L) on primary culture suckling rat osteoblasts can promote the activity of ALP,the OD mv is concentration-dependent, the experimental groups and compared with the control groups were considered significant at a value of P<0.05.3.The suckling rat osteoblasts are culture with intermittent rhPTH1-34(10-9mol/L) interacts SIM(10-8、10-7、10-6mol/L)on primary culture suckling rat osteoblast, compared with every SIM group that corresponded to, the activity of ALP obviously increase,it’s also concentration-dependent and is considered significant at a value of P<0.05.4.Different dose of SIM (10-8、10-7、10-6mol/L) on primary culture suckling rat osteoblasts,can increase the expression of OPG mRNA,reduce the expression of RANKL mRNA,the value is considered significant at a value of P<0.05.5.Compared to the SIM,intermittent rhPTH1-34(10-9mol/L) interacts with Simvastatin (10-8、10-7、10-6mol/L) on primary culture suckling rat osteoblast,the expression of OPG mRNA is obviously up-regulated than the SIM alone and the RANKL is down-regulated.the experimental groups and compared with the control groups were considered significant at a value of P<0.05.Conclusions:rhPTH1-34interacts with SIM show additive effects at the differentiation and metabolism of osteoblasts.