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中国临床药理学与治疗学 ›› 2010, Vol. 15 ›› Issue (9): 1002-1007.

• 基础研究 • 上一篇    下一篇

肝星状细胞氧化应激模型的构建、检测与分析

王启钊1, 吕颖慧1, 李招发1, 江楠2, 刁勇1, 许瑞安1   

  1. 1华侨大学分子药物学研究所·分子药物教育部工程研究中心,泉州 362021,福建;
    2嘉兴市妇幼保健院生殖医学中心,嘉兴 314000,浙江
  • 收稿日期:2010-08-18 修回日期:2010-09-12 出版日期:2010-09-26 发布日期:2020-09-17
  • 通讯作者: 许瑞安,男,博士、教授、博导,研究方向:重大疾病的基因治疗。Tel: 0595-22690952, E-mail: ruianxu@hqu.edu.cn
  • 作者简介:王启钊,男,博士、助理研究员,研究方向:肺癌、肝硬化的基因治疗。Tel: 0595-22690838, E-mail: lyh4444@hqu.edu.cn
  • 基金资助:
    福建科技厅重点项目(2010Y0036); 华侨大学科研基金 (09BS516)

Construction, detection and analysis of the oxidative stress models of hepatic stellate cells

WANG Qi-zhao1, LV Ying-hui1, LI Zhao-fa1, JIANG Nan2, DIAO Yong1, XU Rui-an1   

  1. 1Institute of Molecular Medicine, Huaqiao University; Molecular Medicine Engineering Research Center, Ministry of Education, Quanzhou 362021, Fujian, China;
    2Reproductive Medicine Centre, Jiaxing Maternal and Child Health Hospital, Jiaxing 314000,Zhejiang,China
  • Received:2010-08-18 Revised:2010-09-12 Online:2010-09-26 Published:2020-09-17

摘要: 目的: 建立更为全面的肝星状细胞体外氧化应激模型,为检验药物在肝纤维化氧化应激中发挥的具体作用提供有效工具。方法: 分别用Fe-NTA/AA及过氧化氢作用于大鼠原代肝星状细胞及人肝星状细胞系LX-2细胞以造成不同类型的氧化应激反应,以超氧化物阴离子荧光探针检测细胞内产生的超氧化物水平,并分别用结晶紫染色或MTT检验来检测不同氧化刺激物对两种肝星状细胞体外增殖性的影响。结果: 铁过载及过氧化氢都会引起两种细胞产生氧化应激反应,铁过载可加速原代肝星状细胞激活,对已完全活化的LX-2细胞产生明显毒性作用;过氧化氢则对两种细胞都会造成明显的细胞毒性作用。结论: 处于不同激活状态的肝星状细胞对不同氧化刺激物的反应存在差异,因而在检验肝纤维化药物有效性时,应根据不同的病理、生理需求选择恰当的细胞及氧化应激模型。

关键词: 铁过载, 过氧化氢, 氧化应激, 肝星状细胞, 肝纤维化

Abstract: AIM: To develop correct in vitro oxidative stress models of hepatic stellate cells for the analysis of anti-fibrosis drugs.METHODS: Two representative oxidative models in the liver, the iron-overload model and the H2O2 model were used to generate different oxidative-stress reactions on the primary rat hepatic stellate cells and the immortal cell line of LX-2 cells. The level of intracellular superoxide was detected using dihydroethidium probe and the proliferation status was tested by crystall violet staining and the MTT assay, respectively.RESULTS: Both the iron-overload model and the H2O2 model could generate oxidative reactions in the two kinds of cells. Iron-overload treatment could accelerate the activation of primary hepatic stellate cells, but exerted significant cytotoxicity to LX-2 cells. H2O2 led to completely cytotoxicity to both kinds of cells.CONCLUSION: Hepatic stellate cells at different activation phase would have discrepant responses to the stimulation of different oxidation, thus suitable type of cells and appropriate oxidative-stress related models should be chosen in order to analyse the anti-fibrosis drugs meeting with different physiological and pathological requirement.

Key words: Iron-oxidative, Hydrogen peroxide, Oxiditive stress, Hepatic stellate cells, Liver fibrosis

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