贵州医药2025,Vol.49Issue(1) :3-6.

miR-199a调控Sirt1对SH-SY5Y细胞氧化损伤的影响

Effects of miR-199a in regulation of Sirt1 on oxidative damage of SH-SY5Y nerve cells

万静枝 王婷 廖勇
贵州医药2025,Vol.49Issue(1) :3-6.

miR-199a调控Sirt1对SH-SY5Y细胞氧化损伤的影响

Effects of miR-199a in regulation of Sirt1 on oxidative damage of SH-SY5Y nerve cells

万静枝 1王婷 2廖勇1
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作者信息

  • 1. 三峡大学附属仁和医院,湖北宜昌 443001
  • 2. 武汉科技大学医学院,湖北武汉 430065
  • 折叠

摘要

目的 探讨miR-199a调控Sirt1对人神经母细胞瘤细胞(SH-SY5Y)氧化损伤的影响.方法 采用H2O2(1 200 μmol/L)分别刺激体外培养的SH-SY5Y细胞3、6、9、12、15 h,MTT法检测细胞活力,DCFA-DA探针法检测细胞内ROS水平;miR-199a模拟子和抑制物分别转染SH-SY5Y细胞48 h,Western blot检测Sirt1蛋白的表达;miR-199a模拟子和抑制物分别转染SH-SY5Y细胞48 h后采用H2O2(1 200 μmol/L)刺激12 h,生化法检测细胞超氧化物歧化酶活性和丙二醛含量.结果 不同时间H2O2刺激SH-SY5Y细胞时细胞活力,细胞内超氧化物歧化酶含量升高,且呈现时间依赖性;miR-199a模拟子可抑制SH-SY5Y细胞Sirt1蛋白表达,miR-199a抑制物增加SH-SY5Y细胞Sirt1蛋白表达;当SH-SY5Y细胞处于氧化损伤时,miR-199a模拟子可降低细胞超氧化物歧化酶活性和升高丙二醛含量,miR-199a抑制物增加细胞活力、降低细胞内超氧化物歧化酶含量、升高超氧化物歧化酶活性和降低丙二醛含量.结论 当SH-SY5Y细胞处于氧化损伤状态时,miR-199a调控Sirt1进一步加重SH-SY5Y细胞氧化损伤.

Abstract

Objective To investigate the effects of miR-199a in regulation of Sirt1 on oxidative damage of SH-SY5Y nerve cells.Methods Cultured SH-SY5Y nerve cells were stimulated with H2O2(1 200 μmol/L)for 3 h,6 h,9 h,12 h and 15 h respectively.Cell viability was detected by MTT assay and intracellular ROS levels were de-tected by DCF-DA probe.SH-SY5Y nerve cells were transfected with miR-199a mimics and inhibitors for 48 h,the expression of Sirt1 protein was detected by Western blot.SH-SY5Y nerve cells were transfected with miR-199a mimics and inhibitors for 48 h respectively and stimulated with H2O2(1 200 μmol/L)for 12 h.The activity of su-peroxide dismutase(SOD)and the content of malondialdehyde(MDA)were detected by enzyme biochemical meth-od.Result When SH-SY5Y nerve cells were stimulated by H2O2 at different time,the cell viability and intracellular ROS content increased in a time-dependent manner.The miR-199a mimicon inhibited Sirt1 protein expression in SH-SY5Y neurons,the miR-199a inhibitor increased Sirt1 protein expression in SH-SY5Y neurons.When SH-SY5Y nerve cells were in a state of oxidative damage,miR-199a mimics decreased SOD content and increased MDA content,while miR-199a inhibitor increased cell vitality,decreased intracellular ROS content,increased SOD content and decreased MDA content.Conclusion When SH-SY5Y nerve cells are in the state of oxidative damage,the regulation of Sirt1 by miR-199a further aggravated the oxidative damage of SH-SY5Y nerve cells.

关键词

氧化损伤/人神经母细胞瘤细胞/miR-199a/Sirt1

Key words

miR-199a/Oxidative damage/SH-SY5Y neurons/Sirt1

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出版年

2025
贵州医药
贵州省医药卫生学会办公室

贵州医药

影响因子:0.493
ISSN:1000-744X
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