shRNA-interfered of Nrf2 reveals a critical role for Keap1-Nrf2 signaling pathway during effects of zearalenone induced oxidative stress in IPEC-J2 cells.

IF 2.4 2区 农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE Animal Bioscience Pub Date : 2025-02-01 Epub Date: 2024-08-26 DOI:10.5713/ab.24.0368
Qun Cheng, Shu Zhen Jiang, Li Bo Huang, Wei Ren Yang
{"title":"shRNA-interfered of Nrf2 reveals a critical role for Keap1-Nrf2 signaling pathway during effects of zearalenone induced oxidative stress in IPEC-J2 cells.","authors":"Qun Cheng, Shu Zhen Jiang, Li Bo Huang, Wei Ren Yang","doi":"10.5713/ab.24.0368","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>This study aims to verify the protective effect of the Kelch-like ECH-associated protein1 (Keap1)-nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathways by studying the effect of plasmids containing Nrf2-small hairpin RNA (shRNA) interference down-regulation of Nrf2 on zearalenone (ZEA)-induced intestinal porcine epithelial cells (IPEC-J2) oxidative stress.</p><p><strong>Methods: </strong>We constructed an IPEC-J2 model that interferes with Nrf2 expression, set blank (control), negative control group (Sh-control), positive control group (Sh-Nrf2), and added 10, 20, and 40 μmol/L ZEA experimental group (Sh-Nrf2+ZEA10, Sh-Nrf2+ZEA20, and Sh-Nrf2+ZEA40).</p><p><strong>Results: </strong>The study results showed that, compared with the Sh-Nrf2 group, ZEA significantly increased the apoptosis rate of IPEC-J2 in a time- and dose-dependent manner. Compared with the Sh-Nrf2 group, the activities of total superoxide dismutase and glutathione peroxidase and relative expressions of Keap1 at mRNA and protein level in the Sh-Nrf2+ZEA20 and Sh-Nrf2+ZEA40 groups were significantly reduced, the malondialdehyde level, and the fluorescence intensity around and within the nucleus of reactive oxygen species and Nrf2, and the relative expressions of Nrf2, quinone oxidoreductase 1, and hemeoxygenase 1 at mRNA and protein level significantly increased.</p><p><strong>Conclusion: </strong>These results further prove that interfering with the expression of Nrf2 in IPEC-J2 cells affected the activation of the Keap1-Nrf2 signaling pathway and reduced the ability of cells to resist ZEA-induced oxidative stress. Therefore, the Keap1-Nrf2 signaling pathway had an important protective effect in ZEA-induced intestinal oxidative stress.</p>","PeriodicalId":7825,"journal":{"name":"Animal Bioscience","volume":" ","pages":"303-315"},"PeriodicalIF":2.4000,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11725749/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Animal Bioscience","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.5713/ab.24.0368","RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/8/26 0:00:00","PubModel":"Epub","JCR":"Q1","JCRName":"AGRICULTURE, DAIRY & ANIMAL SCIENCE","Score":null,"Total":0}
引用次数: 0

Abstract

Objective: This study aims to verify the protective effect of the Kelch-like ECH-associated protein1 (Keap1)-nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathways by studying the effect of plasmids containing Nrf2-small hairpin RNA (shRNA) interference down-regulation of Nrf2 on zearalenone (ZEA)-induced intestinal porcine epithelial cells (IPEC-J2) oxidative stress.

Methods: We constructed an IPEC-J2 model that interferes with Nrf2 expression, set blank (control), negative control group (Sh-control), positive control group (Sh-Nrf2), and added 10, 20, and 40 μmol/L ZEA experimental group (Sh-Nrf2+ZEA10, Sh-Nrf2+ZEA20, and Sh-Nrf2+ZEA40).

Results: The study results showed that, compared with the Sh-Nrf2 group, ZEA significantly increased the apoptosis rate of IPEC-J2 in a time- and dose-dependent manner. Compared with the Sh-Nrf2 group, the activities of total superoxide dismutase and glutathione peroxidase and relative expressions of Keap1 at mRNA and protein level in the Sh-Nrf2+ZEA20 and Sh-Nrf2+ZEA40 groups were significantly reduced, the malondialdehyde level, and the fluorescence intensity around and within the nucleus of reactive oxygen species and Nrf2, and the relative expressions of Nrf2, quinone oxidoreductase 1, and hemeoxygenase 1 at mRNA and protein level significantly increased.

Conclusion: These results further prove that interfering with the expression of Nrf2 in IPEC-J2 cells affected the activation of the Keap1-Nrf2 signaling pathway and reduced the ability of cells to resist ZEA-induced oxidative stress. Therefore, the Keap1-Nrf2 signaling pathway had an important protective effect in ZEA-induced intestinal oxidative stress.

查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
ShRNA干扰Nrf2揭示了Keap1-Nrf2信号通路在玉米赤霉烯酮诱导的IPEC-J2细胞氧化应激效应中的关键作用。
研究目的本研究旨在通过研究含有Nrf2-小发夹RNA(shRNA)的质粒干扰下调Nrf2对玉米赤霉烯酮(ZEA)诱导的猪肠上皮细胞(IPEC-J2)氧化应激的影响,验证Kelch样ECH相关蛋白1(Keap1)-核因子红细胞2相关因子2(Nrf2)信号通路的保护作用:方法:构建干扰Nrf2表达的IPEC-J2模型,设置空白对照组(Control)、阴性对照组(Sh-control)、阳性对照组(Sh-Nrf2)和添加10、20、40 μmol/L ZEA的实验组(Sh-Nrf2+ZEA10、Sh-Nrf2+ZEA20、Sh-Nrf2+ZEA40):研究结果表明,与 Sh-Nrf2 组相比,ZEA 能显著提高 IPEC-J2 的凋亡率,且呈时间和剂量依赖性。与Sh-Nrf2组相比,Sh-Nrf2+ZEA20组和Sh-Nrf2+ZEA40组T-SOD和GSH-PX的活性以及Keap1在mRNA和蛋白水平上的相对表达量明显降低,MDA水平、ROS和Nrf2核周围和核内的荧光强度以及Nrf2、Nqo1和Ho1在mRNA和蛋白水平上的相对表达量明显增加:这些结果进一步证明,干扰 Nrf2 在 IPEC-J2 细胞中的表达会影响 Keap1-Nrf2 信号通路的激活,降低细胞抵抗 ZEA 诱导的氧化应激的能力。因此,Keap1-Nrf2信号通路在ZEA诱导的肠道氧化应激中具有重要的保护作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 去求助
来源期刊
Animal Bioscience
Animal Bioscience AGRICULTURE, DAIRY & ANIMAL SCIENCE-
CiteScore
5.00
自引率
0.00%
发文量
223
审稿时长
3 months
期刊最新文献
Effect of dietary supplementation of rubber seed kernel pellet on feed utilization, rumen fermentation, fatty acid profiles and health status in swamp buffalo. Chi-circ_0009659 modulates goat intramuscular adipocyte differentiation through miR-3431-5p/STEAP4 axis. Comparative population genomics analysis for chicken body sizes using genome-wide single nucleotide polymorphisms. Effect of dietary energy levels on productivity, fat deposition, and biochemical parameters of Woorimatdag1 breeder pullets. Effects of fig and pineapple powder on metabolite compounds and health-promoting properties in fermented goat meat sausage.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1