益骨胶囊含药血清对大鼠成骨细胞IGF-ImRNA及其蛋白表达的影响

Effect of YIGU capsule on IGF-I mRNA and protein expression in rat osteoblasts in vitro

目的 :观察益骨胶囊含药血清对SD大鼠成骨细胞 (OB)增殖和胰岛素样生长因子I(IGF -I)mRNA及其蛋白表达的影响。方法 :(1)将 4 0只 12月龄SD雌性大鼠分为益骨胶囊灌胃组 (高、中、低剂量 )和蒸馏水对照组 ,制备含药血清和对照血清 ,并确定最佳灌胃剂量和最佳血清添加浓度 ;(2 )分离、培养新生大鼠成骨细胞 ,传代后分为两组 (含药血清组和对照组 ) ,分别在培养 4 8、72和 96h时做相关指标的测定 ,采用MTT法测定OB增殖 ,RT -PCR法检测IGF -ImRNA的相对表达量 ,ELISA法检测培养上清中IGF -I的浓度。结果 :在 4 8、72和 96h时 ,益骨胶囊含药血清组OB增殖明显高于对照组 (P <0 0 1) ,OB表达IGF -ImRNA和蛋白的量明显高于对照组 (P <0 0 1或P <0 0 5 )。结论 :益骨胶囊含药血清具有促进成骨细胞增殖、表达IGF -ImRNA和蛋白的作用 。

AIM: The effects of YIGU capsule on proliferation and IGF-I mRNA protein expressions in osteoblasts were studied. METHODS: (1) Forty 12-month old Sprague-Dawley female rats were divided randomly into four groups (YIGU capsule high dose group, medium dose group and low dose group; saline group), the drug-containing serum and control serum were prepared. (2) The new-born Sprague-Dawley rat osteoblasts were cultured with different YIGU capsule drug-containing serum at different concentrations and different exposure time. MTT method was used to observe proliferation of osteoblasts. (3) RT-PCR method was used to measure the relative IGF-I mRNA levels and ELISA method was used to measure IGF-I secretion at different exposure time. (4) ELISA method was used to measure IGF-I secretion at different exposure time. RESULTS: (1) Proliferation of osteoblasts was more than the control groups after 48, 72 and 96 h, respectively (P<0.01); (2) The relative IGF-I mRNA levels and IGF-I protein expression were higher than those in control group after 48, 72 and 96 h, respectively (P<0.01 or P<0.05). CONCLUSIONS: It was suggested that YIGU capsule drug-containing serum promoted proliferation, IGF-I mRNA and protein expression. These results may be parts of the mechanisms of YIGU capsule to prevent and treat osteoporosis. [