丹参对人成骨细胞骨钙素生成及钙离子浓度的影响

摘要

目的:研究丹参对于人成骨细胞骨钙素生成及细胞内钙离子浓度的影响.方法:用48孔培养板培养人成骨细胞(hFOB 1.19),分为5个实验组和1个对照组,细胞培养2 d后,实验组分别加入0.05%,0.1%,0.15%,0.2%,0.25%的丹参注射液,对照组不加丹参,再培养3 d后用酶联免疫吸附试验(ELISA)检测各孔培养液中的BGP(Bone Gla Protein,osteocalcin)含量.将人成骨细胞分种于15个培养瓶中,各组5瓶,实验组分为低浓度组和高浓度组,培养3 d后,实验组中分别加入0.1%,0.15%的丹参注射液,对照组不加丹参,再次培养3 d后,用钙离子荧光探针Fluo-3/AM标记后,采用流式细胞术检测人成骨细胞中钙离子的浓度.结果:用不同浓度丹参干预后人成骨细胞BGP的生成量高于对照组(P<0.01),0.15%的丹参浓度作用最强,同样实验组加入不同浓度丹参注射液后人成骨细胞中的钙离子浓度明显高于对照组(P<0.01),也成剂量依赖性.结论:丹参能促进人成骨细胞生成骨钙素,并使细胞内钙离子浓度升高,说明丹参能促进人成骨细胞成熟并提高其活性,从而证明丹参有促进骨形成的能力.%This study was aimed to determine the production of bone Gla protein (BGP or osteocalcin) and the change of calcium ion concentration in human osteoblast (hFOB1.19). Human osteoblasts were cultivated in a 48-well plate, which were assigned into five experimental groups and one control group. After two days cultivation, Salvia miltiorrhiza injection with different concentrations were added in five experimental groups. The concentration was 0.05%, 0.1%, 0.15%, 0.2% and 0.25%, respectively. But there was no S. miltiorrhiza in the control group. After three more days, enzyme-linked immuno sorbent assay (ELISA) was used to detect BGP expression. The human osteoblasts were cultivated in 15 culture flasks, which were assigned into experimental groups and control group. After three days cultivation, 0.1% S. miltiorrhiza injection was added into the experimental group one, and 0.15% S. miltiorrhiza injection was added into the experimental group two. After three more days, fluorescent probe Fluo-3/AM was used to mark the osteoblasts. The flow cytometry was used to detect calcium ion concentration in the osteoblasts. The results showed that all experimental groups with different concentration of salvia miltiorrhiza produced more BGP than the control group (P < 0.01). The most effective concentration was 0.15%. After the intervention of S. miltiorrhiza, the calcium ion concentration in the osteoblasts was higher, and the calcium ion concentration in the osteoblasts with 0.15% salvia miltiorrhiza was higher than osteoblasts with 0.1% S. miltiorrhiza (P < 0.01). It was concluded that Chinese medicine S. miltiorrhiza can promote human osteoblasts in the production of more BGP, and make human osteoblasts have higher calcium ion concentration. Therefore, S. miltiorrhiza can stimulate the osteoblast maturity, and promote bone formation and mineralization.