从ERα/RANK通路探讨淫羊藿苷抑制破骨细胞分化作用  

Mechanism of Icariin in Inhibiting Osteoclast Differentiation Via ERα/ RANK Signal Pathway

作　　者：李伟娟[1] 谢保平 石丽颖 李劲平[1] 曾英[2] 张杰[1] 雷晓明[2] 

机构地区：[1]中南大学药学院,长沙410013 [2]湖南中医药大学第一附属医院,长沙410007

出　　处：《中国实验方剂学杂志》2017年第7期121-126,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(81273816);中南大学研究生自主探索创新项目(2016zzts496)

摘　　要：目的:通过检测淫羊藿苷对破骨细胞诱导分化过程中雌激素受体α(estrogen receptorα,ERα),核转录因子-κB受体(receptor activator of nuclear factor-κB,RANK)mRNA表达的影响,探讨淫羊藿苷抑制破骨细胞分化作用机制。方法:50μg·L^(-1)核因子κB受体活化因子配体(receptor activator of nuclear factor-κB ligand,RANKL)体外诱导RAW264.7细胞分化成破骨细胞,利用抗酒石酸酸性磷酸酶(TRAP)染色和骨吸收陷窝对破骨细胞进行鉴定。用不同浓度的淫羊藿苷(1×10-5,1×10-6,1×10-7mol·L^(-1))分别处理5 d和9 d后,进行TRAP染色和骨吸收陷窝分析,处理6 d后,利用逆转录-聚合酶链式反应(RT-PCR)检测基质金属蛋白酶-9(MMP-9),组织蛋白酶K(cathepsin-K,CK),TRAP,ERα,RANK mRNA表达水平,用雌激素受体阻断剂ICI182780阻断雌激素受体通路验证淫羊藿苷对ERα/RANK通路的调节作用。结果:与RANKL诱导组比较,淫羊藿苷可以显著抑制破骨细胞形成数量和骨吸收陷窝数(P<0.05,P<0.01),同时显著下调破骨细胞MMP-9,CK,TRAP,RANK mRNA表达水平(P<0.05,P<0.01),上调ERαmRNA表达水平(P<0.05)。ICI182780可抑制淫羊藿苷上调破骨细胞的ERαmRNA表达水平和下调RANK mRNA表达水平的作用(P<0.05)。结论:淫羊藿苷能抑制RANKL诱导RAW264.7向破骨细胞分化和骨吸收活性,该作用可能通过上调破骨细胞的ERαmRNA表达进而下调RANK mRNA表达实现的。Objective： To explore the mechanism of icariin in inhibiting the osteoclast differentiation via detecting the mRNA expression levels of estrogen receptor a （ERα） and receptor activator of NF-κB （RANK） in osteoclast induced from RAW264.7. Method： RAW264.7 ceils were induced into osteoblasts with receptor activator of nuclear factor KB ligand （RANKL） （50 μg·L^-1）, and were identified by tartrate resistant acid phosphatase （TRAP） staining and bone resorption pit formation. After RAW264.7 ceils were treated with different concentrations （1×10^-5,1×10^-6,1×10^-7mol·L^-1） of icariin for 5 days and 9 days, TRAP staining, and bone resorption pit formation analysis were conducted. After treatment for 6 days, the gene expression levels of （MMP-9） ,cathepsin-K （CK）, TRAP, RANK and ERα in osteoclasts were detected by RT-PCR. ICI182780, an estrogen receptor antagonist, was used to invalidate the regulatory effect of icariin on ERa/RANK pathway. Result： Icariin could significantly inhibit the number of osteoclast formation and number of bone resorption pits （P 〈0. 05）, significantly down-regulate the gene expression levels of MMP-9, CK, TRAP and RANK in the osteoclasts （P 〈0.05）, and up-regulate the gene expression of ERa. ICI182780 could inhibit the icariin in up- regulating the gene expression level of ERα and down-regulating the gene expression level of RANK. Conclusion： Icariin could inhibit osteoclast differentiation and bone absorption function, which may be achieved via increasing ERα mRNA expression and decreasing RANK mRNA expression.

关 键 词：淫羊藿苷 RAW264.7细胞 破骨细胞 雌激素受体Α 核因子κB受体 

分 类 号：R285.5[医药卫生—中药学]