由“从瘀论治”理论探讨三七中槲皮素对破骨细胞分化的自噬调控机制  

To explore quercetin in Panax Notoginseng on autophagy regulation mechanism of osteoclast differentiation based on the theory of“treatment from stasis”

作　　者：刘昭明 关智宇 蒋太平 刘志伦 李成蹊 LIU Zhaoming;GUAN Zhiyu;JIANG Taiping;LIU Zhilun;LI Chengxi(Guizhou University of Traditional Chinese Medicine,Guiyang 550002;Guizhou Provincial Hospital of Traditional Chinese Medicine,Guiyang 550001,China)

机构地区：[1]贵州中医药大学,贵州贵阳550002 [2]贵州省中医医院,贵州贵阳550001

出　　处：《中国骨质疏松杂志》2022年第12期1740-1744,1749,共6页Chinese Journal of Osteoporosis

基　　金：国家自然科学基金资助项目(81960813)。

摘　　要：目的探究槲皮素对破骨细胞分化自噬调控机制的影响。方法本实验分为对照组、RANKL组、RANKL+槲皮素组。通过CCK8检测摸索槲皮素的处理浓度,按照上述分组采用Raw 264.7细胞诱导7 d后进行TRAP染色验证模型成功后,加槲皮素干预48 h进行TRAP染色,通过qPCR检测TRAP、CTSK、NFATC-1、ATG5、p62、mTOR、Beclin-1基因表达。结果根据CCK8结果选择槲皮素1 mg/L浓度;TRAP染色结果显示RANKL组破骨细胞显著增多,槲皮素干预后破骨细胞减少;qPCR结果显示模型组TRAP、P62、CTSK、NFATC-1基因表达上升,ATG5、Beclin-1、mTOR基因表达下降,槲皮素干预组TRAP、CTSK、NFATC-1、ATG5、p62、mTOR、Beclin-1基因均下降。电镜观察显示槲皮素干预后破骨细胞自噬小体数量减少。结论槲皮素可以通过降低自噬基因表达水平抑制破骨细胞的分化。Objective To explore the effect of quercetin on autophagy regulation mechanism of osteoclast differentiation.Methods The experiment was divided into control group,RANKL group and RANKL+quercetin group.CCK8 was used to detect the concentration of quercetin.After 7 days of induction with Raw 264.7 cells according to the above grouping,the model was verified by TRAP staining.After 48 hours of quercetin intervention,TRAP staining was performed,and qPCR was used to detect the gene expression of TRAP,CTSK,NFATC-1,ATG5,p62,mTOR and Beclin-1.Results Quercetin 1 mg/L was selected according to CCK8 result.TRAP staining showed that osteoclasts increased significantly in RANKL group,and osteoclasts decreased after quercetin intervention.qPCR result showed that the gene expression of TRAP,P62,CTSK and NFATC-1 increased,while the gene expression of ATG5,Beclin-1 and mTOR decreased in the model group.The gene expression of TRAP,CTSK,NFATC-1,ATG5,p62,mTOR and Beclin-1 decreased in the quercetin intervention group.Electron microscopic observation showed that the number of autophagy bodies in osteoclasts decreased after quercetin intervention.Conclusion Quercetin can inhibit osteoclast differentiation by reducing the expression level of autophagy genes.

关 键 词：三七 槲皮素 破骨分化 自噬 骨质疏松症 

分 类 号：R285.5[医药卫生—中药学]