低氧对破骨细胞活性的影响及其机制

摘要

Objective:To observe the effects of hypoxia on activity of osteoclasts and to investigate the possible mechanism. Methods: The osteoblasts and osteoclasts were obtained from new - laid rabbits by mechanical separation and cultured in vitro. Osteoclasts were identified by tartrate -resistant acid phosphatase (TRAP) staining. Osteoclasts resorptive pits on bone slice cultured in normoxia (20% oxygen concentration) and hypoxia (3% oxygen concentration) groups were compared. The purified osteoclasts were added to the 3 rd generation osteoblasts. The jointly cultured cells were divided into normoxia group and hypoxia group. Real - time reverse transcription PCR (RT-PCR) were used to detect the expressions of RANK, OPG, RANKL, TRAP and CtsK at 24, 48, 72 and 96 hours after culture. Results: Compared with normoxia group, the expression of RANK, RANKL, CtsK and TRAP Mrna were increased but the expression of OPG Mrna was decreased. Conclusion: Hypoxia could exert trophic effects on the activity of osteoclasts by activating the upstream signaling pathway of TRAP and CtsK.%目的:观察低氧对乳兔破骨细胞分化、活性的影响,并研究其可能的机制.方法:培养乳兔破骨细胞、成骨细胞,TRAP染色鉴定破骨细胞,比较破骨细胞在常氧(含氧量20％)及低氧(含氧量3％)条件下骨吸收陷窝面积.破骨细胞加入到第3代成骨细胞中分别于常氧和低氧条件下进行共培养,第24、48、72、96 h检测RANK、OPG、RANKL、TRAP及CtsK mRNA的表达.结果:各时间点低氧组RANK、RANKL、CtsK、TRAP mRNA表达均高于常氧组；OPG mRNA的表达低于常氧组组,具有统计学差异(P＜0.05或P＜0.01).结论:低氧可激活破骨细胞TRAP和CtsK基因的上游信号,增强破骨细胞活性.