中文摘要：

目的通过观察补肾健脾中药健骨颗粒含药血清对体外破骨细胞CAⅡ、CK、MMP-9mRNA表达的影响,进一步揭示健骨颗粒有效防治绝经后骨质疏松症的作用机制。方法用M-CSF和RANKL诱导破骨前体细胞RAW264.7分化为成熟破骨细胞,alamar Blue法检测破骨细胞血清干预的最佳浓度,于破骨细胞培养第7天开始干预,分别用25%浓度的健骨颗粒含药血清和25%浓度的生理盐水血清干预,培养48小时后抽提总RNA,采用逆转录-聚合酶链反应（RT-PCR）、实时荧光定量SYBR GREEN法检测CAⅡ、CK、MMP-9mRNA表达,并取骨片行甲苯胺蓝染色,计算分析骨陷窝数和骨吸收面积,同时运用氧化显色法检测破骨细胞培养液中TRAP含量,ELISA法检测骨磨片培养液中CTx含量。结果健骨颗粒含药血清组破骨细胞CAⅡ、CK、MMP-9mRNA表达水平明显低于生理盐水血清组（P〈0.05）,含药血清组骨磨片的骨陷窝数和骨吸收面积以及TRAP和CTx含量均低于生理盐水血清组（P〈0.05）。结论健骨颗粒能有效抑制破骨细胞CAⅡ、CK、MMP-9mRNA表达,降低破骨细胞功能活性,抑制破骨细胞对骨基质的分解,从而抑制骨吸收。

英文摘要：

Objective To observetheeffect of Jiangu granulecontaining serum on themRNA expression of CA II,CK,and MMP-9 in osteoclasts,and to reveal themechanism of Jiangu granulein prevention and treatment of postmenopausal osteoporosis.Methods Matureosteoclasts wereinduced from osteoclast precursor RAW264. 7 cells with M-CSF and RANKL. Theoptimal concentration of serum intervention was detected with Alamar Bluemethod. On theseventh day of culture,thecells weretreated with 25% normal salinecontaining serum or 25% Jiangu granulecontaining serum,respectively. After 48 hours,total RNA was extracted and then reversely transcripted using RT-PCR. Real-timefluorescencequantitativeSYBR GREEN method was applied to detect CA II,CK,and MMP-9 mRNA expression. Toluidinebluestaining bonechip was used to analyzeboneresorption lacuna and area. Oxidation color method was used to detect TRAP content in culturefluid. ELISA method was used to detect CTx content in culturebonegrinding tablets. Results CA II, CK, and MMP-9 mRNA expression levels in Jiangu granulegroup weresignificantly lower than thosein normal salinegroup（ P〈0. 05）. Boneresorption lacuna and area on grinding tablets and levels of TRAP and CTx in Jiangu granulegroup werelower than thosein normal salinegroup（ P〈0. 05）. Conclusion Jiangu granuleinhibits CA II,CK,and MMP- 9 mRNA expression in osteoclasts effectively,reduces functional activity of osteoclasts,and inhibits resorption of bonematrix by osteoclasts.