| ObjectiveTo explore the effect of rhPTH(1-34) on the proliferation and expression of Collagen I andOsterix on rat osteoblast in vitro.And investigate the possible mechanism of action on osteoblasts,provide further theoretical basis for clinical medication.Method1,Culture rat osteoblasts,and observe their morphology and function.2,Cultured rat osteoblast in vitro were divided into control group and five differentrhPTH(1-34) dosages ranging from 10-11 M to 10-7M of drug group,with 24 hours for anincubation cycle and cultured with rhPTH (1-34) at the first 12 hours .After 48 hours,cellproliferation was assessed by MTT colorimetric assay,the expression of Collagen I andOsterix mRNA in rat osteoblast were examined by semiquantitive RT-PCR.3,Statistical analysis: Data are presented as x±s. All statisticalanalysis is performed withWindows SPSS13.0. The differents between the groups are determined by One-wayANOVE.The differences between random two groups are determined by LSD.Differencesare considered significant at a value of P<0.05.Result1,Most of cultured rat osteoblasts were elongated spindle-shaped, ovate-triangular orirregular polygon, a round or oval nuclei, center or slant at a side, surrounding cells toprotuberant connected. Alkaline phosphatase staining was positive, cytoplasm was blackparticles, Alizarin red staining was dark red nodules.2,Cultured rat osteoblasts was promoted by different concentrationsof rhPTH (1-34), eachOD are increased, rhPTH(1-34) can induce the proliferation of osteoblast especially at 10-9M, in does-dependent manner.The experimental groups and compared with the controlgroups were considered significant at a value of P < 0.05.3,Semiquantative RT-PCR examination revealed: rhPTH (1-34) promoted Collagen I andOsterix mRNA expression, especially at 10-9 M, in does-dependent manner.The experimentalgroups and compared with the control groups were considered significant at a valueof P < 0.05.Conclusion1,rhPTH (1-34) promotes proliferation of cultured rat osteoblasts.2,rhPTH (1-34) promotes expression of Collagen I and Osterix mRNA. 3,rhPTH (1-34) may regulate the expression of Collagen I and Osterix mRNA to promoteosteoblast proliferation,differentiation and maturation. |
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