26-失碳-8-氧代-α-芒柄花萜醇对体外培养成骨细胞活性的影响

为研究玉柏石松提取物26-失碳-8-氧代-α-芒柄花萜醇(26-NO-Ono)对成骨细胞活性的影响,采用MTT检测不同浓度26-NO-Ono(3.33,6.66,13.32,26.64μmol/L)对成骨细胞增殖率,碱性磷酸酶(ALP)试剂盒检测成骨细胞内ALP活性,荧光定量PCR检测成骨细胞骨相关基因表达.结果表明:26-NO-Ono给药1d可促进成骨细胞增殖,给药3d可促进成骨细胞ALP活性.26-NO-Ono处理3d和9d会抑制骨涎蛋白(BSP)、I型胶原蛋白(Col-I)以及骨钙素蛋白(OCN)的基因表达;处理6d会促进上述基因的表达.26-NO-Ono长期处理(6d和9d)可以抑制骨桥蛋白(OPN)的基因表达,说明26-NO-Ono对成骨细胞的成骨活性的影响呈时间依赖性,剂量依赖性和细胞分化状态依赖性.

Effects of 26-NO-Ono on Activity of Cultured Osteoblasts in vitro

To study the effects of 26-nor-8-oxo-α-onocerin(26-NO-Ono),an extract from Lycopodium obscurum L.,on the activity of cultured osteoblasts in vitro,different concentrations(3.33,6.66,13.32,26.64μmol/L) of 26-NO-Ono were given to cultured osteblasts.MTT assay,alkaline phosphate(ALP) kit,real-time PCR were applied to measure the proliferation rate,activity of ALP and expression of bone-related genes in treated osteoblasts.Results showed that treatment with 26-NO-Ono for 1d can promote the osteoblast proliferation rate and 3 d can enhance the ALP activity.Treatment with 26-NO-Ono for 3 d and 9 d can inhibit the expression of bone-related genes such as BSP,Col-I and OCN in osteoblasts,while treatment for 6 d can stimulate the above 3 genes.Meanwhile,treatment with 26-NO-Ono for 6 d and 9 d could inhibit the expression of OPN gene.Our results indicated that the effect of 26-NO-Ono on osteogenic activity of osteoblast is time-dependent,concentration-dependent and differentiation state-dependent.