杯苋甾酮通过拟雌激素样作用抑制破骨细胞分化  

Cyasterone inhibits osteoclast differentiation by estrogen-like effect

作　　者：谢景春 赵玥 黄立慧[3] 程禄萍 金奇[4] 谢保平 XIE Jing-chun;ZHAO Yue;HUANG Li-hui;CHENG Lu-ping;JIN Qi;XIE Bao-ping(Department of Burns and Plastics Surgery,People’s Hospital of Yudu County,Ganzhou Jiangxi 342399;Key Laboratory of Prevention and Treatment of Cardiovascular and Cerebrovascular Diseases,Ministry of Education,Gannan Medical University,Ganzhou Jiangxi 341000;Department of Informatics,The First Affiliated Hospital of Gannan Medical University,Ganzhou Jiangxi 341000;Department of Research,Gannan Medical University,Ganzhou Jiangxi 341000)

机构地区：[1]于都县人民医院烧伤整形外科,江西赣州342399 [2]赣南医学院心脑血管疾病防治教育部重点实验室,江西赣州341000 [3]赣南医学院第一附属医院信息科,江西赣州341000 [4]赣南医学院科研处,江西赣州341000

出　　处：《中南药学》2022年第9期2046-2051,共6页Central South Pharmacy

基　　金：江西省教育厅科学技术研究项目(No.GJJ180809;No.GJJ201510;No.GJJ15094);赣南医学院校级课题(No.YB201901);赣南医学院心脑血管疾病防治教育部重点实验室开放基金(No.XN202025)。

摘　　要：目的 探讨杯苋甾酮的拟雌激素样作用及其抑制破骨细胞分化的作用机制。方法 采用CCK-8试剂检测杯苋甾酮对MCF-7和RAW264.7细胞增殖的影响;用酒石酸酸性磷酸酶染色(TRAP)检测杯苋甾酮对破骨细胞分化的影响,并用雌激素受体阻断剂ICI182780(ICI)阻断;用qRT-PCR和Western blot检测杯苋甾酮对MCF-7和破骨细胞中ERα mRNA和蛋白水平表达以及破骨细胞分化相关基因表达的影响;用分子对接验证杯苋甾酮与ERα的相互作用关系。结果 杯苋甾酮(5、10 μmol·L^(-1))可显著促进MCF-7细胞的增殖以及ERα mRNA和蛋白水平的表达;且实验剂量下杯苋甾酮可显著抑制破骨细胞分化以及TRAP和RANK的表达,上调ERα基因与蛋白水平的表达;ICI可以阻断杯苋甾酮对MCF-7细胞增殖和破骨细胞分化的影响,上调TRAP和RANK的表达;分子对接发现杯苋甾酮与17-β雌二醇对ERα共同作用位点是在Glu-353和His-524。结论 杯苋甾酮具有拟雌激素样作用,并可通过ERα信号通路抑制破骨细胞分化。Objective To determine whether cyasterone has an estrogen-like effect and the mechanism of action of cyasterone in inhibiting osteoclast differentiation.Methods The effect of cyasterone on the proliferation of MCF-7 cells and RAW264.7 cells was detected by CCK-8 reagent.The effect of cyasterone on the osteoclast differentiation was detected by TRAP staining and blocked by ICI182780 (ICI).Western blot and qRT-PCR were used to detect the effect of cyasterone on the expression of ERα mRNA and protein in MCF-7 and osteoclast cells,including the expression of osteoclast differentiation-related genes,and computer simulation docking to confirm the interaction of cyasterone and ERα.Results Cyasterone (5,10 μmol·L^(-1)) obviously promoted the proliferation of MCF-7 cells and the expression of ERα mRNA and protein.Cyasterone at the experimental dose greatly inhibited the osteoclast differentiation,as well as the expression of TRAP and RANK,and up-regulated the expression of ERα gene and protein.ICI blocked the effect of cyasterone on the proliferation of MCF-7 and osteoclast differentiation,and up-regulated the expression of TRAP and RANK.Computer simulation showed that the co-action sites of cyasterone and 17-β estradiol on ERα were Glu-353 and His-524.Conclusion Cyasterone has an estrogen-like effect and inhibits the osteoclast differentiation through the ERα signaling pathway.

关 键 词：破骨细胞 杯苋甾酮 雌激素受体 骨质疏松症 

分 类 号：R285[医药卫生—中药学]