目的 探讨加减青娥方抑制骨质疏松的作用机制。方法 在体外培养的小鼠破骨前体RAW264.7细胞中加入不同浓度的加减青娥方提取物,利用抗酒石酸酸性磷酸酶(TRAP)染色法检测核因子κ B受体活化因子配体(RANKL)诱导的RAW264.7细胞分化为破骨细胞的数量及其活性的影响;利用荧光定量PCR(RT-PCR)法检测RANKL诱导的RAW264.7细胞分化为破骨细胞雌激素受体(ER)mRNA表达。结果 加减青娥方可抑制RANKL诱导的RAW264.7细胞分化为破骨细胞,该作用可能是通过调控ERα mRNA的表达实现的。结论 加减青娥方可通过调控ERα基因的表达,抑制破骨细胞的分化、增殖,从而实现增加骨密度、防治骨质疏松的作用。

Objectives To explore the molecular biology mechanism of Modified Qing'e Formula in the treatment of postmenopausal osteoporosis (PMOP). Methods The extractions of Modified Qing'e Formula with different doses were added into the osteoclast precursors RAW264.7 cultured in vitro. Then the number and activity of osteoclasts differentiated from osteoclast precursors RAW264.7 induced by receptor activator of nuclear factor κ B factor ligand (RANKL) were tested with tartrate-resistant phosphatase (TRAP) staining; and the estrogen receptor (ER) mRNA expressions of those osteoclasts were examined with fluorescence quantitative Real-time RT-PCR. Results Modified Qing'e Formula inhibited the RANKL-induced RAW264.7 cells to differentiate into osteoclasts, which might be achieved by regulating the expression of ERα mRNA. Conclusion Modified Qing'e Formula could suppress the differentiation and proliferation of osteoclasts to increase bone density and to prevent and treat PMOP via regulating the expression of ERα mRNA.

国家自然科学基金青年科学基金项目(81202800);高等学校博士学科点专项科研基金(20111210120014);天津市中医药管理局科研课题(11025)