褪黑素通过Nrf-2途径抑制高铁诱导成骨细胞的凋亡
Melatonin inhibits the apoptosis induced by excess iron via activation of Nrf-2 signal pathway in osteoblasts
  
DOI:10.3969/j.issn.1006.7108.2015.08.001
中文关键词:  褪黑素  铁  成骨细胞  Nrf-2信号通路  细胞凋亡
英文关键词:Melatonin  Iron  Osteoblasts  Nrf-2 signaling pathway  Cell apoptosis
基金项目:国家自然科学基金项目(81170808 ,81471094);辽宁省教育厅基金(L2013301 );沈阳市科学技术基金(F12-S77-4-47)
作者单位
刘俊华 杨茂伟* 孟洪正 施鹏旭 易远历 刘霏 中国医科大学附属第一医院骨外科,沈阳110001 
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中文摘要:
      目的 探讨Nrf-2途径在褪黑素抑制高铁诱导成骨细胞凋亡中的作用。方法 用细胞贴壁法培养成骨细胞 (hFOB1. 19)后,将枸橼酸铁铵(400μmol/ L)和褪黑素(100μmol/L)加人细胞培养基,培养24 h后,Annexin V-FITC/PI流式细胞学检测细胞凋亡水平;DCFH-DA荧光探针检测细胞内ROS水平;Western blot检测凋亡相关蛋白线粒体内Bax和caspase- 3,以及Nrf-2途径相关蛋白Nrf-2和HO-1。结果 高浓度枸橼酸铁铵干预成骨细胞后,与对照组相比,细胞内ROS水平增高,枸橼酸铁铵处理后凋亡增加,凋亡相关蛋白,线粒体内Bax表达增加,caspase-3表达增加,同时,Nrf-2途径相关蛋白Nrf-2及 HO-1表达增加。而褪黑素可以进一步增加细胞内的Nrf-2及HO-1水平,同时降低细胞内ROS水平,细胞凋亡率和凋亡相关蛋白表达明显降低。结论 褪黑素可以通过Nrf-2途径抑制成骨细胞的氧化应激水平,从而降低高铁引起的成骨细胞凋亡。
英文摘要:
      Objective To investigate the effect of melatonin on the apoptosis of hFOB1. 19 cells induced by excess iron. Methods hFOB1. 19 cells were treated with ferric ammonium citrate (400μmol/L) and melatonin (100μmol/L) for 24 h. The apoptosis rate and the level of ROS were detected using flow cytometry. Proteins associated with apoptosis like Bax and caspase-3 and associated with Nrf-2 signal pathway like Nrf-2 and HO-1 were determined using Western blotting. Results The level of ROS and apoptosis rate increased after intervention with excess iron. The levels of Bax in the mitochondria and cleaved caspase-3 in the cytosol increased. However,after the pretreatment with melatonin,the level of ROS,apoptosis rate,and expression of apoptosis associated proteins decreased,and the expression of Nrf-2 and HO-1 increased. Conclusion Melatonin inhibits the level of oxidation in osteoblasts via Nrf-2 signal pathway,resulting in the reduction of apoptosis induced by excess iron.
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