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中国临床药理学与治疗学 ›› 2008, Vol. 13 ›› Issue (6): 620-626.

• 基础研究 • 上一篇    下一篇

化学性缺氧致大鼠脑细胞[Ca2+]i和脑红蛋白表达变化及阿司匹林干预研究

程菊, 谢坤, 侯敏, 白德成   

  1. 甘肃省新药临床前研究重点实验室, 兰州大学基础医学院人体解剖与组织胚胎学研究所, 兰州 730000, 甘肃
  • 收稿日期:2007-12-06 修回日期:2008-05-27 发布日期:2020-10-14
  • 通讯作者: 白德成,男,教授,硕士生导师,研究方向:神经生物学。Tel:13088758222 E-mail:bdc@lzu.edu.cn
  • 作者简介:程菊,女,医学硕士,讲师,主要从事脑缺氧方面的研究。Tel:13321209385 E-mail:chengj@lzu.edu.cn
  • 基金资助:
    甘肃省新药临床前研究重点实验室开放及基金资助项目(GSKFKT-0703);甘肃省自然科学基金暨中青年科技基金项目(3ZS061-A25-089);兰州大学医学科研基金资助项目(010602)

Chemical hypoxia induced [Ca2+]i and NGB alteration and aspirin intervention

CHENG Ju, XIE Kun, HOU Min, BAI De-cheng   

  1. Key Laboratory of Pre-clinical Study for New Drugs of Gansu Province, Institute of Anatomy and Histology and Embryology, Basic Medical College, Lanzhou University, Lanzhou 730000, Gansu, China
  • Received:2007-12-06 Revised:2008-05-27 Published:2020-10-14

摘要: 目的: 观察化学性缺氧状况下大鼠脑细胞[Ca2+]i和脑红蛋白(NGB)表达的变化及阿司匹林(ASA)对其变化的干预,并初步探讨其作用机制。方法: 连二亚硫酸钠致原代培养大鼠脑细胞缺氧,用荧光探针Flou-3负载和NGB荧光免疫组化技术双标,激光共聚焦显微镜动态观察。结果: [Ca2+]i和NGB表达与缺氧持续时间有关,随缺氧时间的延长,荧光强度随之增高;经ASA预防性干预,两种荧光强度增高缓慢,其增高程度明显低于单纯缺氧组;治疗性干预,用药早期两种荧光强度增高明显,其增高程度与单纯缺氧组变化相近,后期则增高缓慢,其增高程度明显低于单纯缺氧组。[Ca2+]i浓度和NGB表达呈正相关。结论: ASA对原代培养大鼠脑细胞化学性缺氧损伤具有保护作用,其作用可能与ASA抑制细胞内Ca2+超载有关;NGB的表达可能是受到Ca2+-CaM复合物的调节;ASA预防性用药的早期保护作用明显优于治疗性用药。

关键词: 阿司匹林, 脑, 细胞培养, 缺氧, [Ca2+]i, 脑红蛋白

Abstract: AIM: To observe changes of intracellular free calcium levels ([Ca2+]i) and neuroglobin (NGB) expression when rats′brain cells were exposed to chemical hypoxia, and the influence of aspirin (ASA) on changes as well as to study its mechanism primarily.METHODS: Sodium dihionite was used to deprive the oxygen in culture medium, and confocal laser scanning microscope with double fluorescence labeling methods were used to measure the changes of [Ca2+]i and NGB expression in rats′brain cells.Flou-3/AM and Cy-3 were used to indicate calcium concentration and NGB expression respectively.RESULTS: [Ca2+]i and NGB expression are related to the time of hypoxia.Compared with the normal control group, average fluorescent intensity of [Ca2+]i and NGB expression in the simple hypoxia group increased gradually with the time of the exposure in hypoxia.Moreover, there is the positive correlation between [Ca2+]i and NGB expression.[Ca2+]i and NGB expression in the ASA pretreated group increased slowly, the increase degree is significantly lower than that in the simple hypoxia group.[Ca2+]i and NGB expression in the ASA treated group were increased significantly in early stage, the increase degree is not significant diffexence from the simple hypoxia group.At 120 min, [Ca2+]i and NGB expression in the ASA treated group increased slowly, the increase degree is significantly lower than that in the simple hypoxia group.CONCLUSION: ASA has a protective effect on hypoxic injury of rats′ brain cells, which may be attribute to its inhibiting calcium overload, and NGB expression is regulated by Ca2+-CaM complex.In addition, the neuroprotective effects of ASA pretreated group were better than ASA treated group in early stage.

Key words: aspirin, brain, cell culture, hypoxica, [Ca2+]i, neuroglobin

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