基础医学与临床 ›› 2014, Vol. 34 ›› Issue (9): 1193-1198.

• 研究论文 • 上一篇    下一篇

Sulfiredoxin-1通过调节Peroxiredoxins表达提高大鼠星形胶质细胞抗氧化作用

周杨,周云川,巫静娴,喻姗姗,赵涌   

  1. 重庆医科大学
  • 收稿日期:2014-05-30 修回日期:2014-07-09 出版日期:2014-09-05 发布日期:2014-09-02
  • 通讯作者: 赵涌 E-mail:blys01@163.com
  • 基金资助:
    国家自然科学基金资助项目

Sulfiredoxin-1 enhances the antioxidant effects of astrocytes by regulating the expression of peroxiredoxins

  • Received:2014-05-30 Revised:2014-07-09 Online:2014-09-05 Published:2014-09-02
  • Contact: Yong ZHAO E-mail:blys01@163.com

摘要: 目的 探讨内源性抗氧化小分子蛋白Sulfiredoxin1(Srxn1)对H2O2 诱导的星形胶质细胞的抗氧化作用以及对内源性氧化应激防御系Peroxiredoxins 的调节作用。方法Srxn1干扰慢病毒载体转染体外培养的大鼠原代星形胶质细胞;构建H2O2氧化应激模型,采用LDH 方法检测细胞损伤程度,用超氧化物歧化酶(SOD)分析细胞内氧化应激状态;Western blot检测Srxn1的表达与PrdxⅠ~Ⅳ和Prdx-SO2/3H(过氧化Prdx)活性的关系。 结果1) sh-Srxn1干扰后,Srxn1蛋白水平下调59.2%(P<0.01), 基因水平下调63.6%(P<0.01)。2)干扰Srxn1增加H2O2 后LDH漏出率(P<0.01)并使氧化应激损伤显著加重、使PrdxⅠ~Ⅳ的表达降低,Prdx-SO2/3H的表达显著增高(P<0.01或 P<0.05)。结论Srxn1减轻H2O2诱导的星形胶质细胞的氧化应激损伤,并可能是通过调节Prdxs的活性来完成的。

关键词: Sulfiredoxin1, 干扰, 星形胶质细胞, H2O2 , Peroxiredoxin

Abstract: Objective To explore the possible mechanisms of Sulfiredoxin1 (Srxn1) for its antioxidant effects in astrocytes in response to oxidative stress induced by H2O2. Methods 1)Incubation and purification of astrocytes from the cerebral cortex of SD rats. 2)The lentiviral interference vector carrying Srxn1 or negative control(NC) shRNA were transduced into primary rat cortical astrocytes. And the knockdown efficiency of Srxn1 was measured by Real-time QPCR and Western blot assays. 3)H2O2 was treated after Srxn1 knockdown. Using LDH assays to measure cell damage. The intracellular oxidative stress state was estimated by superoxide dismutase(SOD) kit. The expression of Srxn1 and the activity of PrdxⅠ-Ⅳ and Prdx-SO2/3H were measured by WB. Results 1)WB revealed that the expression of Srxn1 was decreased to 40.8%(P<0.01)and Real-time QPCR showed that mRNA of Srxn1 was decreased to 36.4%(P<0.01)after knockdown of Srxn1. 2)Knockdown Srxn1 could increase the leakage rate of LDH to 23.4%(P<0.01)and decrease the level of SOD to 53.1%(P<0.01) after exposed to H2O2. 3)Exposed to H2O2 resulted in an increase in Srxn1 to 137.3%(P<0.01). The expression of PrdxⅠ-Ⅳ can be increased by H2O2. However, it could be decreased after knockdown of Srxn1. Conclusion Srxn1 could alleviate oxidative stress injury for astrocytes and involved in Prdxs’ activity.

Key words: Sulfiredoxin1, interference, astrocytes, H2O2, Peroxiredoxin

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