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作 者:齐鹏坤[1,2] 侯德才[2] 吕艳芳 王一品[2] QI Pengkun;HOU Decai;LYU Yanfang;WANG Yipin(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,Liaoning,China;The Fourth People's Hospital of Shenyang,Shenyang 110031,Liaoning,China)
机构地区:[1]辽宁中医药大学,辽宁沈阳110847 [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
出 处:《中华中医药学刊》2024年第6期86-90,I0016,I0017,共7页Chinese Archives of Traditional Chinese Medicine
基 金:国家中医药管理局中医药行业专项(2014070020);辽宁省教育厅科学研究经费项目(L202032);沈阳市科技计划项目(22-321-34-10)。
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Bcl-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。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
分 类 号:R274[医药卫生—中医骨伤科学]
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