紫菀酮对体外破骨细胞分化和活性的影响及作用机制研究  

Effects of shionone on d ifferentiation and activity of osteoclasts in vitro and its mechanism of action

作　　者：苏珮茹 罗香雅 余丽娜 曾春平 周琳 SU Peiru;LUO Xiangya;YU Lina;ZENG Chunping;ZHOU Lin(The Fifth Affiliated Hospital of Guangzhou Medical University,Guangzhou 510700,Guangdong,China;School of Public Health,North China University of Science and Technology,Tangshan 063210,Hebei,China)

机构地区：[1]广州医科大学附属第五医院,广东广州510700 [2]华北理工大学公共卫生学院,河北唐山063210

出　　处：《中医正骨》2023年第6期24-35,共12页The Journal of Traditional Chinese Orthopedics and Traumatology

基　　金：广东省基础与应用基础研究基金项目(2019A1515110723)。

摘　　要：目的:探讨紫菀酮对体外破骨细胞分化和活性的影响及作用机制。方法:①分析紫菀酮对核因子κB受体活化因子配体(receptor activator of nuclear factorκB ligand,RANKL)诱导Raw 264.7细胞向破骨细胞分化的影响。将Raw 264.7细胞分为空白对照组、阳性对照组、紫菀酮1μmol·L^(-1)干预组、紫菀酮2.5μmol·L^(-1)干预组、紫菀酮5μmol·L^(-1)干预组和紫菀酮10μmol·L^(-1)干预组,空白对照组加入完全培养基,阳性对照组加入含25 ng·mL^(-1)RANKL的完全培养基,其余各组分别加入含有相应浓度紫菀酮及25 ng·mL^(-1)RANKL的完全培养基,培养5 d后进行抗酒石酸酸性磷酸酶(tartrateresistant acid phosphatase,TRAP)染色,统计各组破骨细胞数;②分析紫菀酮对Raw 264.7细胞活力的影响。将Raw 264.7细胞分为空白对照组、紫菀酮1μmol·L^(-1)干预组、紫菀酮2.5μmol·L^(-1)干预组、紫菀酮5μmol·L^(-1)干预组和紫菀酮10μmol·L^(-1)干预组,空白对照组加入完全培养基,其余各组分别加入含有相应浓度紫菀酮的完全培养基,分别培养48 h、120 h后测定各组细胞活力;③分析紫菀酮对破骨细胞纤维状肌动蛋白环形成的影响。将Raw 264.7细胞分为空白对照组、阳性对照组、紫菀酮5μmol·L^(-1)干预组和紫菀酮10μmol·L^(-1)干预组,空白对照组加入完全培养基,阳性对照组加入含25 ng·mL^(-1)RANKL的完全培养基,紫菀酮5μmol·L^(-1)干预组和紫菀酮10μmol·L^(-1)干预组分别加入含有相应浓度紫菀酮及25 ng·mL^(-1)RANKL的完全培养基,培养5 d后采用鬼笔环肽和4’,6二脒基2苯基吲哚染色,观察各组破骨细胞纤维状肌动蛋白环形成情况。④分析紫菀酮对核因子κB(nuclear factorκB,NFκB)转录活性的影响。将稳定转染pGL4.32[luc2P/NFκBRE/Hygro]载体的RAW 264.7细胞分为空白对照组、阳性对照组和紫菀酮10μmol·L^(-1)干预组,空白对照组及阳性对照组加入完全培养基,紫菀酮10μmol·L^Objective:To explore the effects of shionone on differentiation and activity of osteoclasts in vitro and its mechanism of action.Methods:①To analyze the effects of shionone on the differentiation of Raw 264.7 cells into osteoclasts induced by receptor activator of nuclear factorκB ligand(RANKL).The Raw 264.7 cells were divided into blank control group,positive control group,1μmol/L shionone intervention group,2.5μmol/L shionone intervention group,5μmol/L shionone intervention group and 10μmol/L shionone intervention group.The Raw 264.7 cells in blank control group were cultured in complete culture medium,the ones in positive control group were cultured in complete culture medium supplemented with 25 ng/mL RANKL,and the ones in the rest groups were cultured in complete culture medium supplemented with 25 ng/mL RANKL and shionone at concentration of 1,2.5,5 and 10μmol/L respectively.After 5day culture,tartrateresistant acid phosphatase(TRAP)staining was performed,and the number of osteoclasts in each group was counted.②To analyze the effects of shionone on the viability of Raw 264.7 cells.The Raw 264.7 cells were divided into blank control group,1μmol/L shionone intervention group,2.5μmol/L shionone intervention group,5μmol/L shionone intervention group and 10μmol/L shionone intervention group.The Raw 264.7 cells in blank control group were cultured in complete culture medium,and the ones in the other groups were cultured in complete culture medium supplemented with shionone at concentration of 1,2.5,5 and 10μmol/L respectively.After 48and 120hour culture,the viability of Raw 264.7 cells were detected in each group.③To analyze the effects of shionone on the formation of fibrous actin rings(Factin rings)in osteoclasts.The Raw 264.7 cells were divided into blank control group,positive control group,5μmol/L shionone intervention group and 10μmol/L shionone intervention group.The Raw 264.7 cells in blank control group were cultured in complete culture medium,the ones in positive control group were cu

关 键 词：骨质疏松 破骨细胞 细胞分化 紫菀酮 体外研究技术 信号传导 NFκB 组织蛋白酶K 腺苷三磷酸酶 抗酒石酸酸性磷酸酶 NFATC转录因子 

分 类 号：R285.5[医药卫生—中药学]