中华中医药学刊2024,Vol.42Issue(6) :86-90,后插16-后插17.DOI:10.13193/j.issn.1673-7717.2024.06.016

龟鹿二仙胶诱导破骨细胞凋亡的机制研究

Mechanism Study on Apoptosis of Osteoclasts Induced by Guilu Erxian Jiao(龟鹿二仙胶)

齐鹏坤 侯德才 吕艳芳 王一品
中华中医药学刊2024,Vol.42Issue(6) :86-90,后插16-后插17.DOI:10.13193/j.issn.1673-7717.2024.06.016

龟鹿二仙胶诱导破骨细胞凋亡的机制研究

Mechanism Study on Apoptosis of Osteoclasts Induced by Guilu Erxian Jiao(龟鹿二仙胶)

齐鹏坤 1侯德才 2吕艳芳 3王一品2
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作者信息

  • 1. 辽宁中医药大学,辽宁沈阳 110847;辽宁中医药大学附属医院,辽宁沈阳 110032
  • 2. 辽宁中医药大学附属医院,辽宁沈阳 110032
  • 3. 沈阳市第四人民医院,辽宁沈阳 110031
  • 折叠

摘要

目的 探讨不同浓度龟鹿二仙胶对破骨细胞凋亡影响及相关机制.方法 RAW 264.7细胞培养传代,用M-CSF+RANKL诱导小鼠单核细胞RAW 264.7细胞株分化成破骨细胞,获得破骨细胞;CCK8法、TUNEL染色、流式细胞法检测不同浓度的龟鹿二仙胶对破骨细胞凋亡的影响;Real-time PCR法、Western blot法检测破骨细胞凋亡相关分子Bel-2、Bax、Cytochrome C的表达;经龟鹿二仙胶及PI3K/AKT通路抑制剂LY294002干预后,通过流式细胞法、Western blot法检测破骨细胞凋亡率及凋亡相关蛋白AKT的变化.结果 不同浓度的龟鹿二仙胶(GLEXJ高、中、低)均能抑制破骨细胞的增殖活性,提高破骨细胞的凋亡率,提高凋亡相关分子Bax/Bcl-2的比值以及Cytochrome C的表达,其中,以中浓度GLEXJ作用72 h后影响最显著;经中浓度龟鹿二仙胶,以及PI3K/AKT通路抑制剂LY294002的干预,明显抑制了 PI3K/AKT通路相关蛋白AKT蛋白的磷酸化,破骨细胞总凋亡率显著上升.结论 龟鹿二仙胶通过抑制PI3K/AKT通路活化作用而促进破骨细胞凋亡.

Abstract

Objective To investigate the effects of different concentration of Guilu Erxian Jiao(龟鹿二仙胶)on osteoclast ap-optosis and osteoclast relevant mechanism.Methods RAW264.7 cells were cultured and passaged and RAW264 in mouse mono-cytes was induced using M-CSF+RANKL 7 cell lines to differentiate into osteoclasts and obtain osteoclasts.CCK8 method,TUNEL staining and low cytometric analysis were used to detect the effects of different concentrations of Guilu Erxian Jiao on os-teoclast apoptosis.The expressions of apoptosis-related molecules Bcl-2,Bax and Cytochrome C were detected by Real-time PCR and Western blot.After the intervention of Guilu Erxian Jiao and phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)pathway inhibitor LY294002,the apoptosis rate of osteoclasts and apoptosis-related protein AKT were detected by flow cytometry and Western blot.Results Different concentrations of Guilu Erxian Jiao(high,medium,low dose)could inhibit prolifer-ative activity of osteoclasts,improve the apoptosis rate,the ratio of Bax/Bcl-2 and Cytochrome C expression.But the effect of Guilu Erxian Jiao at medium concentration for 72 h was the most significant.After the intervention of medium concentration of Guilu Erxian Jiao and PI3K/AKT pathway inhibitor LY294002,the phosphorylation of PI3K/AKT pathway related protein AKT protein was significantly inhibited,and the total apoptosis rate of osteoclasts was significantly increased.Conclusion Guilu Erxian Jiao can promote apoptosis of osteoclasts by inhibiting PI3K/AKT pathway activation.

关键词

中医中药/龟鹿二仙胶/PI3K/AKT/破骨细胞/凋亡

Key words

traditional Chinese medicine/Guilu Erxian Jiao(龟鹿二仙胶)/PI3K/AKT/osteoclasts/apoptosis

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基金项目

国家中医药局中医药行业科研专项(2014070020)

辽宁省教育厅科学研究经费项目(L202032)

沈阳市科技计划(22-321-34-10)

出版年

2024
中华中医药学刊
中华中医药学会 ,辽宁中医药大学

中华中医药学刊

CSTPCD北大核心
影响因子:1.007
ISSN:1673-7717
参考文献量27
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