淫羊藿甙促进体外培养成骨细胞骨成形蛋白-2及Osterix mRNA表达
Icarrin increases bone morphogenetic protein 2 and Osterix expression in rat osteoblast
目的 观察淫羊藿甙对体外培养成骨细胞增殖、分化以及骨成形蛋白-2(BMP-2)、成骨细胞特异性转录因子Osterix mRNA表达的影响,探讨淫羊藿抗骨质疏松的作用机制.方法 体外原代培养用酶消化法获得的新生SD大鼠颅盖骨成骨细胞.在培养液中分别加入不同浓度的淫羊藿甙,用RT-PCR法测定成骨细胞BMP-2、Osterix、碱性磷酸酶(ALP)和Ⅰ型胶原(Col Ⅰ)mRNA表达.将淫羊藿甙(10 ng/ml)与BMP-2单克隆抗体单独或共同加入培养液中,培养48 h,用RT-PCR测定成骨细胞Osterix mRNA表达水平的变化.结果 1、10、100 ng/ml淫羊藿甙均可促进BMP-2、Osterix 、ALP和Col Ⅰ mRNA表达(P<0.05),以10 ng/ml组作用最显著.淫羊藿甙和BMP-2单克隆抗体混合组Osterix mRNA表达下调,与淫羊藿甙组比较,差异有统计学意义(P<0.05),而与BMP-2抗体组比较,差异无统计学意义(P>0.05).结论 淫羊藿甙能提高成骨细胞中ALP和Col Ⅰ mRNA的表达水平,从而促进成骨细胞增殖与分化,并通过提高BMP-2的表达水平来促进成骨细胞Osterix的基因表达,从而诱导骨形成,发挥抗骨质疏松作用.
更多Objective To observe the effect of icarrin on proliferation and differentiation of rat osteoblasts cultured in vitro as well as the expressions of bone morphogenetic protein-2 (BMP-2) and Osterix in rat osteoblast.Methods Calvarial osteoblasts were obtained from newborn (< 24 h) SD rats by trypsin-collagenase digestion.The culture medium with different icarrin concentrations and the second generation osteoblasts were mixed.The mRNA expressions of BMP-2,Osterix,ALP,and Col Ⅰ were detected by semiquantative RT-PCR.Then the second generation osteoblasts were cultured in the medium containing icariin (10 ng/ml) with or without BMP-2 antibody.After48 h,RT-PCR was used to estimate the mRNA expression of Osterix.Results In comparison with the control group,the mRNA expressions of BMP-2,Osterix,ALP,Col Ⅰ were increased in a dose-dependent manner (P<0.05) with a maximal effect at the concentration of 10 ng/ml.The mRNA expression of Osterix treated with the mixture of icarrin and BMP-2 antibody was significant decreased(P<0.05),and compared with the group of icarrin,however,the difference between the BMP-2 group and the mixture group was not statistically significant (P > 0.05).Conclusions Icarrin stimulates proliferation and differentiation of cultured osteoblast in vitro by increasing the mRNA expression of ALP and Col Ⅰ.Icarrin could stimulate Osterix gene expression by enchancing BMP-2 gene expression,finally,it could induce bone formation and prevent and/or treat osteoporosis.
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