猪m6A去甲基化酶FTO、ALKBH5基因表达水平与大肠杆菌F18感染抗性的关系分析

doi:10.16431/j.cnki.1671-7236.2020.09.031

摘要/Abstract

摘要： 为了探讨猪6-甲基腺嘌呤（m⁶A）去甲基化酶mRNA表达水平与仔猪大肠杆菌F18抗性的关系，本研究运用实时荧光定量PCR方法检测m⁶A去甲基化酶FTO和ALKBH5基因在35日龄苏太猪断奶仔猪大肠杆菌F18抗性型和敏感型个体十二指肠和空肠组织中的mRNA表达差异，同时分别利用F18ab、F18ac大肠杆菌菌体刺激和内毒素LPS诱导猪小肠上皮细胞（IPEC-J2），检测FTO、ALKBH5基因表达变化。结果表明，FTO、ALKBH5基因在抗性型个体十二指肠和空肠中的表达量均极显著或显著高于敏感型个体（P<0.01，P<0.05）；不同F18大肠杆菌菌体刺激IPEC-J2细胞后，FTO基因表达水平均无显著变化（P>0.05），而ALKBH5基因在F18ac刺激后表达量显著上调（P<0.05）；LPS诱导4 h时，FTO和ALKBH5基因表达量均显著上调（P<0.05）。本研究在细胞和个体水平上初步验证并发现了m⁶A去甲基化酶FTO和ALKBH5基因表达水平与大肠杆菌感染仔猪密切相关，为今后深入研究RNA去甲基化修饰在仔猪细菌性腹泻调控中的作用机制提供了理论基础。

关键词: 猪; m⁶A甲基化; FTO基因; ALKBH5基因; 大肠杆菌

Abstract: To investigate the relationship between the mRNA expression level of m⁶A demethylase and E.coli F18 resistance in piglets,Real-time quantitative PCR was used to detect the mRNA expression differences of m⁶A demethylase FTO and ALKBH5 genes in the duodenum and jejunum tissues of E.coli F18-resistant and -sensitive individuals from 35-day Sutai weaned piglets.In E.coli F18ab,F18ac bacteria-stimulated and endotoxin LPS-induced porcine small intestinal epithelial cells (IPEC-J2),the expression levels of FTO and ALKBH5 genes were detected,respectively.The results showed that the expression levels of FTO and ALKBH5 genes in the duodenum and jejunum of resistant individuals were extremely significantly or significantly higher than those of sensitive individuals (P<0.01,P<0.05),and the expression of FTO gene were not significantly changed in E.coli F18 bacteria-stimulated IPEC-J2 cells (P>0.05),but the expression levels of ALKBH5 gene were significantly up-regulated after F18ac stimulation (P<0.05).After LPS induction for 4 hours,the expression levels of FTO and ALKBH5 genes showed significant up-regulation in IPEC-J2 cells (P<0.05).This study preliminarily verified and indicated that the expression levels of m⁶A demethylases FTO and ALKBH5 genes were closely related to E.coli resistance of piglets at the cellular and individual levels,which will provide a theoretical basis for future in-depth study of the regulation mechanism of RNA demethylation modification on bacterial diarrhea in piglets.

Key words: pigs; m⁶A demethylase; FTO gene; ALKBH5 gene; E.coli

中图分类号:

S828.8

齐晓艺, 杨荔, 吴正常, 包文斌, 吴圣龙. 猪m⁶A去甲基化酶FTO、ALKBH5基因表达水平与大肠杆菌F18感染抗性的关系分析[J]. 中国畜牧兽医, 2020, 47(9): 2960-2967.

QI Xiaoyi, YANG Li, WU Zhengchang, BAO Wenbin, WU Shenglong. Relationship Between Expression Levels of Porcine m⁶A Demethylases FTO and ALKBH5 Genes and Resistance to E.coli F18 Infection[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(9): 2960-2967.

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猪m⁶A去甲基化酶FTO、ALKBH5基因表达水平与大肠杆菌F18感染抗性的关系分析

Relationship Between Expression Levels of Porcine m⁶A Demethylases FTO and ALKBH5 Genes and Resistance to E.coli F18 Infection

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