井冈山大学学报自然科学版

文章摘要

王雨晴,来明名,陈艳,刘永新.过氧化氢体外诱导小鼠胚胎成纤维细胞氧化损伤模型的构建和分析[J].井冈山大学自然版,2022,43(1):57-62

过氧化氢体外诱导小鼠胚胎成纤维细胞氧化损伤模型的构建和分析

ESTABLISHMENT AND ANALYSIS OF NIH-3T3 OXIDATIVE DAMAGE MODEL INDUCED BY H₂O₂ IN VITRO

投稿时间：2021-07-16 修订日期：2021-10-02

DOI：10.3969/j.issn.1674-8085.2022.01.010

中文关键词: 过氧化氢 NIH-3T3细胞氧化损伤丙二醛一氧化氮 γH₂AX

英文关键词: H₂O₂ NIH-3T3 oxdiative damage MDA NO γH₂AX

基金项目:国家自然科学基金项目（81660465）；云南省中药饮片产业发展专项资金（云财建（2019）292号）

作者	单位
王雨晴	大理大学基础医学院, 云南, 大理 671000
来明名	大理大学基础医学院, 云南, 大理 671000
陈艳	大理大学基础医学院, 云南, 大理 671000
刘永新	大理大学基础医学院, 云南, 大理 671000

摘要点击次数: 2186

全文下载次数: 3823

中文摘要:

为给抗氧化损伤药物的开发利用和抗氧化损伤机制的研究提供理论依据和实验基础，本方法构建了过氧化氢（H₂O₂）体外诱导小鼠胚胎成纤维细胞（NIH-3T3）经典的氧化应激损伤模型。应用贴壁细胞培养法培养NIH-3T3细胞，采用不同浓度的H₂O₂处理NIH-3T3细胞不同时间模拟氧化损伤模型，通过CCK-8法观察H₂O₂引起NIH-3T3细胞半数致死率浓度（IC50）和时间；检测丙二醛（MDA）以及一氧化氮（NO）的表达水平；免疫荧光法观察H₂O₂诱导NIH-3T3细胞氧化损伤后细胞核内组蛋白γH₂AX的表达变化。通过与对照组进行分析比较，300 μM H₂O₂处理NIH-3T3细胞2 h后，NIH-3T3细胞死亡率最接近50%。此时，NIH-3T3细胞内MDA和NO水平显著增高，细胞核中组蛋白γH₂AX表达水平也明显提高。研究表明，H₂O₂体外诱导NIH-3T3细胞氧化应激模型的最适条件：300 μM H₂O₂处理2 h。

英文摘要:

In order to provide theoretical and experimental basis for the development and utilization of anti-oxidative damage drugs and the mechanism of anti-oxidative damage, the classical oxidative stress injury model of NIH-3T3 induced by H₂O₂ in vitro was established. NIH-3T3 was cultured according to the adherent cell culture method. NIH-3T3 cells were treated with different concentrations of H₂O₂ for different times to simulate oxidative damage model. CCK-8 test was used to investigate the half maximal inhibitory concentration (IC50) and time of NIH-3T3. The expression levels of MDA and NO were measured according to the detection kits. The expression of γH₂AX was observed by immunofluorescence assay. Compared with the control, the apoptosis rate of NIH-3T3 treated with 300 μM H₂O₂ for 2 hours was almost 50%. At the same time, MDA and NO levels in NIH-3T3 were significantly increased, and the γH₂AX expression in the nucleus was also significantly increased. The results indicated that treatment with 300 μM H₂O₂ for 2 hours was the optimal condition for inducing NIH-3T3 oxidative damage in vitro.

查看全文查看/发表评论下载PDF阅读器

关闭