人工虎骨粉干预大鼠成骨细胞增殖和Ⅰ型胶原的表达

摘要

背景:人工虎骨粉主要用于骨质疏松症、骨折、类风湿关节炎等骨科疾病的治疗,并在临床中取得较好的疗效,但其成骨作用的药物机制尚未明确.目的:观察人工虎骨粉对大鼠成骨细胞增殖和Ⅰ型胶原表达的影响.方法:采用新生SD大鼠颅盖骨组织二次酶消化法培养原代成骨细胞,其中人工虎骨粉干预组细胞培养液的药物终质量浓度分别为10-4,10-5,10-6 g/L.对照组给予不含人工虎骨粉的无血清DMEM细胞培养液.结果与结论:噻唑蓝比色法检测结果显示,人工虎骨粉可显著提高成骨细胞增殖活力(P < 0.05),流式细胞仪检测结果显示,人工虎骨粉可提高S期和M/G2期细胞分布比例,并伴有G0/ G1期细胞比例降低;RT-PCR和ELISA的检测结果显示,各质量浓度人工虎骨粉处理成骨细胞48 h,均可以提高成骨细胞Ⅰ型胶原的表达(P < 0.05).结果表明人工虎骨粉对体外培养的大鼠原代成骨细胞具有促增殖作用并可以促进Ⅰ型胶原的合成和分泌.%BACKGROUND: Bionic tiger-bone powder is mainly used for the treatment of osteoporosis, fracture, rheumatoid arthritis and other bone diseases, and has achieved better clinical efficacy. However, the mechanisms of osteogenesis has not clear yet. OBJECTIVE: To observe the effects of bionic tiger-bone powder on the proliferation of rat osteoblasts and the expression of typeⅠcollagen. METHODS: We cultivate primary osteoblasts with secondary enzyme digestion of the newborn rat skull bone tissue. The Concentrations of culture medium in bionic tiger-bone powder intervention group were 10-4, 10-5 and 10-6 g / L. The control group was injected with the serum-free DMEM culture medium without bionic tiger-bone powder. RESULTS AND CONCLUSION: MTT colorimetric assay results showed that bionic tiger-bone powder could improve osteoblasts proliferation vitality apparently (P < 0.05). Meanwhile the proportion of S phase and M/G2 phase cells was increased but that of G0/G1 phase cells was decreased through flow cytometry. The RT-PCR and ELISA test showed that bionic tiger-bone powder could increase the expression of osteoblasts typeⅠcollagen (P < 0.05) when handling with them in the concentration of 10-6, 10-5 and 10-4 mg/ml for 48 hours. Bionic tiger-bone powder could promote the proliferation of rat primary osteoblasts in vitro, and could promote the synthesis and secretion of typeⅠcollagen.