目的 探討加減青娥方抑制骨質(zhì)疏松的作用機(jī)制。方法 在體外培養(yǎng)的小鼠破骨前體RAW264.7細(xì)胞中加入不同濃度的加減青娥方提取物,利用抗酒石酸酸性磷酸酶(TRAP)染色法檢測核因子κ B受體活化因子配體(RANKL)誘導(dǎo)的RAW264.7細(xì)胞分化為破骨細(xì)胞的數(shù)量及其活性的影響;利用熒光定量PCR(RT-PCR)法檢測RANKL誘導(dǎo)的RAW264.7細(xì)胞分化為破骨細(xì)胞雌激素受體(ER)mRNA表達(dá)。結(jié)果 加減青娥方可抑制RANKL誘導(dǎo)的RAW264.7細(xì)胞分化為破骨細(xì)胞,該作用可能是通過調(diào)控ERα mRNA的表達(dá)實(shí)現(xiàn)的。結(jié)論 加減青娥方可通過調(diào)控ERα基因的表達(dá),抑制破骨細(xì)胞的分化、增殖,從而實(shí)現(xiàn)增加骨密度、防治骨質(zhì)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.

國家自然科學(xué)基金青年科學(xué)基金項(xiàng)目(81202800);高等學(xué)校博士學(xué)科點(diǎn)專項(xiàng)科研基金(20111210120014);天津市中醫(yī)藥管理局科研課題(11025)