摘要
目的:探究知母皂苷BⅡ(TBⅡ)对破骨细胞分化过程中核转录因子-κB受体活化因子配基(RANKL),RANK,原癌基因(C-FOS)基因表达量的影响。方法:利用分子对接软件LeDock对TBⅡ分别与RANKL,RANK和C-FOS进行分子对接打分。RAW264.7细给予可溶性RANKL(sRANKL)干预,设立空白组,sRANKL组(模型组),淫羊藿苷(Ica)组,TBⅡ低剂量组(2μmol·L^-1),TBⅡ中剂量组(4μmol·L^-1),TBⅡ高剂量组(8μmol·L^-1)。相应试剂盒检测破骨细胞分化的标志性酶(TRAP),实时荧光定量聚合酶链式反应(Real-time PCR)检测C-FOS,与其上游RANKL/RANK和下游活化T细胞核因子胞质1型(NFATC1)表达量,以及骨保护素OPG的表达量。结果:分子对接打分结果分别为-11.86,-11.38,-12.34 kcal·mol^-1,TBⅡ能够与RANKL,RANK和C-FOS结合。与正常组比较,模型组TRAP含量显著升高(P<0.01);与模型组比较,各给药组显著降低TRAP含量(P<0.01),且TBⅡ呈剂量依赖性降低TRAP含量。与正常组比较,模型组RANKL,RANK,C-FOS,NFATC1表达量显著升高(P<0.01),OPG表达量显著降低(P<0.01);与模型组比较,各给药组显著降低RANKL,RANK,C-FOS,NFATC1表达量(P<0.01),显著升高OPG表达量(P<0.01)。结论:TBⅡ可能通过调控RANKL/RANK/C-FOS信号通路,抑制破骨细胞前体细胞向破骨细胞的分化,抑制破骨细胞活性,减少骨吸收,改善骨质疏松。
Objective:To explore the effect of anemarrhena asphodeloside BⅡ(TBⅡ)on the expressions of nuclear transcription factor-κB receptor activator factor ligand(RANKL),RANK and C-FOS genes during osteoclast differentiation.Method:Molecular docking software LeDock was used to score the docking of TBⅡ with RANKL,RANK and C-FOS.RAW264.7 was treated with soluble RANKL(sRANKL)and divided into control group,sRANKL group(model group),Icariin(Ica)group,low-dose TBIⅡ group(2μmol·L^-1),medium-dose TBⅡgroup(4μmol·L^-1),and high-dose TBⅡ group(8μmol·L^-1).The corresponding kit was used to detect iconic enzyme(TRAP)of osteoclast differentiation.Total RNA was extracted by trizol method,Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect the expressions of C-FOS,upstream RANKL/RANK and downstream nuclear factor of activated T-cells cytoplasmic 1(NFATC1),and osteoprotegerin OPG.Result:The molecular docking score were-11.86,-11.38,-12.34 kcal·mol-1,and there might be multiple binding sites between TBⅡ as well as RANKL,RANK and C-FOS.Compared with the control group,the content of TRAP in model group increased significantly(P<0.01),and compared with model group,the content of TRAP in each administration group decreased significantly(P<0.01),and TBⅡ decreased the content of TRAP in a dose-dependent manner.Compared with the control group,the expressions of RANKL,RANK,C-FOS and NFATC1 increased(P<0.01),whereas the expression of OPG decreased(P<0.01)in model group.Compared with model group,the expressions of RANKL,RANK,C-FOS and NFATC1 decreased(P<0.01),while the expression of OPG increased(P<0.01)in each administration group.Conclusion:TBⅡ may inhibit the differentiation of osteoclast precursors into osteoclasts,inhibit osteoclast activity,reduce bone resorption and improve osteoporosis by regulating RANKL/RANK/C-FOS signal pathway.
作者
冯萌
党院霞
刘芬
邢菊玲
莫紫欣
罗瑞娇
周欣欣
FENG Meng;DANG Yuan-xia;LIU Fen;XING Ju-ling;MO Zi-xin;LUO Rui-jiao;ZHOU Xin-xin(School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2020年第19期146-152,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81473439)。
