低氧暴露对大鼠骨骼肌L6 细胞铁代谢的影响.docVIP

生理学报 Acta Physiologica Sinica, August 25, 2011, 63(4): 347–352  347  研究论文 低氧暴露对大鼠骨骼肌L6细胞铁代谢的影响 李海洲，刘玉倩*，王海涛，刘树欣，赵斌，杨杰 河北师范大学体育学院，石家庄 050024 摘 要：本文旨在研究低氧暴露对骨骼肌细胞铁代谢的影响。培养大鼠骨骼肌L6细胞，随机分成3组，分别低氧(1% O2)暴露 0、12和24 h。采用同位素55Fe示踪法测定细胞铁摄取和铁释放能力的变化；用流式细胞仪检测细胞内可变铁池(labile iron pool, LIP)；应用蛋白免疫印迹检测转铁蛋白受体1 (transferrin receptor 1, TfR1)、二价金属离子转运体(divalent metal transport- er 1, DMT1)、膜铁转运蛋白1 (ferroportin 1, FPN1)和低氧诱导因子1 (hypoxia-inducible transcription factor-1, HIF-1)的变化。结 果显示，与0 h低氧暴露组相比，12 h低氧暴露组细胞的铁摄取能力明显升高(P 0.05)，铁释放能力显著降低(P 0.01)，LIP 明显升高(P 0.05)。24 h低氧暴露组肌细胞铁摄取、铁释放能力及LIP均明显低于0 h和12 h低氧暴露组(P 0.05 或P 0.01)。与0 h低氧暴露组比较，12 h低氧暴露组HIF-1、DMT1 (IRE)、DMT1 (non-IRE)和TfR1表达显著升高(P 0.05 或P 0.01)，而FPN1表达则显著降低(P 0.01)；24 h低氧暴露组DMT1 (IRE)、DMT1 (non-IRE) 和FPN1表达均低于0和12 h低氧暴 露组(P 0.01)，但TfR1表达却显著增加(P 0.05或P 0.01)。以上结果提示，低氧对骨骼肌铁代谢的影响与暴露时间密切相 关，一定时间的低氧暴露可以提高肌细胞的铁摄取能力，降低铁释放，增加细胞内的铁含量，而长时间的低氧暴露会引起 细胞铁代谢紊乱。 关键词：低氧暴露；L6细胞；低氧诱导因子1；铁 中图分类号：Q74 Effects of hypoxia on iron metabolism of rat L6 skeletal muscle cells LI Hai-Zhou, LIU Yu-Qian*, WANG Hai-Tao, LIU Shu-Xin, ZHAO Bin, YANG Jie Department of Physical Education, Hebei Normal University, Shijiazhuang 050024, China Abstract: The present study was aimed to investigate the effects of hypoxia on iron metabolism of skeletal muscle. Rat L6 skeletal muscle cells were randomly divided into three groups which were exposed to hypoxia (1% O2) for 0, 12, 24 h, respectively. Iron iso- tope tracing method was used to determine iron uptake and release. Iron content of labile iron pool (LIP) was investigated by flow cy- tometry, and the expressions of transferrin receptor 1 (TfR1), divalent metal transporter 1 (DMT1), ferroportin 1 (FPN1), hypoxia-in- ducible transcription factor-1 (HIF-1) in L6 cells were observed by Western blot. The results showed that, compared with 0-hour hypoxia group, 12-hour hypoxia group exhibited significantly increased iron uptake and LIP (P 0.05), as well as decreased iron re- lease (P 0.01). Not only iron uptak