基础医学与临床 ›› 2016, Vol. 36 ›› Issue (3): 321-326.

• 研究论文 • 上一篇    下一篇

铁过载损伤小鼠睾丸支持细胞

李莉1,赵昱2,龚淼3,王慧娟1,高福禄1   

  1. 1. 河北医科大学
    2. 河北医科大学组织学与胚胎学教研室
    3. 河北医科大学组织胚胎学教研组
  • 收稿日期:2015-08-03 修回日期:2015-11-30 出版日期:2016-03-05 发布日期:2016-02-22
  • 通讯作者: 高福禄 E-mail:gfl@mail.hebtu.edu.cn
  • 基金资助:
    基于信号转导通路网络节点研究何首乌饮干预睾丸组织退行性病变的分子机制

Iron overload injures Sertoli cells of mouse

  • Received:2015-08-03 Revised:2015-11-30 Online:2016-03-05 Published:2016-02-22

摘要: 目的 探讨铁过载对体外原代培养的小鼠睾丸支持细胞(SC)的影响。方法 取原代分离培养小鼠睾丸SC纯化并鉴定,加入不同浓度(50、100、200 μmol/L)右旋糖酐铁注射液,培养至24、48和72 h后收集细胞,光学显微镜与流式细胞术鉴定铁过载,流式细胞术检测细胞活性氧(ROS)水平,试剂盒检测丙二醛(MDA)、谷胱甘肽(GSH)和一氧化氮(NO)的含量及超氧化物歧化酶(SOD)、一氧化氮合酶(NOS)含量和谷胱甘肽过氧化物酶(GSH-Px)的活性,Western blot 与免疫细胞化学检测闭锁蛋白(occludin)、雄激素结合蛋白(ABP)、转铁蛋白(TRF)、抑制素(INH)和波形蛋白(VIM)的表达。结果 与对照组相比,铁过载组细胞内ROS、MDA、NO与NOS的含量显著增高(P<0.01),GSH含量、SOD和GSH-Px的活性显著降低(P<0.01),occludin、ABP、TRF、INH和VIM蛋白的表达明显降低(P<0.05)。结论 铁过载可导致体外培养的睾丸支持细胞氧化性损伤并降低其功能。

关键词: 关键词: 铁过载, 支持细胞, 小鼠, ROS, 氧化损伤

Abstract: Objective To investigate the effect of iron overload on sertoli cells. Methods sertoli cells were isolated and cultured by adding iron dextran at different concentrations (50,100,200μmol/L ) for different lengths of time (24,48,72h). Iron overload was observed by microscopy and flow cytometry. ROS production was observed by flow cytometry. GSH and GSH-Px contents were determined by nitrobenzoic acid method.MDA content was measured by thiobarbituric acid way.NO and NOS contents were detected by nitrate reductase way.SOD content was examined by xanthine oxidase method.The expression levels of occluding,ABP,TRF,INH and VIM protein were observed by Western blot and immunocytochemistry. Results Compared with control group, the level of intracellular ROS, the contents of MDA, NO and NOS all significantly increased, with consequent decrease in the contents of SOD, GSH, GSH-Px (P<0.01).The expressions of occluding,ABP,TRF,INH and VIM were also significantly decreased (P<0.05).Conclusion Iron overload can induce oxidative damage of sertoli cells and weaken its function.

Key words: iron overload, Sertoli cell, mouse, ROS, oxidative damage

中图分类号: