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METTL3 通过 TLR2/NF-κB 信号通路调控 CPB2 毒素暴露的 IPEC-J2 细胞的炎症反应。

METTL3 Regulates the Inflammatory Response in CPB2 Toxin-Exposed IPEC-J2 Cells through the TLR2/NF-κB Signaling Pathway.

机构信息

College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China.

Gansu Academy of Agricultural Sciences (CAAS), Lanzhou 730070, China.

出版信息

Int J Mol Sci. 2022 Dec 13;23(24):15833. doi: 10.3390/ijms232415833.

DOI:10.3390/ijms232415833
PMID:36555481
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9781724/
Abstract

beta2 (CPB2) toxin is one of the main pathogenic toxins produced by , which causes intestinal diseases in animals and humans. The N6-methyladenosine (m6A) modification is the most common reversible modification in eukaryotic disease processes. Methyltransferase-like 3 (METTL3) regulates immunity and inflammatory responses induced by the bacterial infections in animals. However, METTL3's involvement in CPB2-treated intestinal porcine epithelial cell line-J2 (IPEC-J2) remains unclear. In the current study, we used methylated RNA immunoprecipitation-quantitative polymerase chain reaction, Western blotting and immunofluorescence assay to determine the role of METTL3 in CPB2-exposed IPEC-J2 cells. The findings revealed that m6A and METTL3 levels were increased in CPB2 treated IPEC-J2 cells. Functionally, METTL3 overexpression promoted the release of inflammatory factors, increased cytotoxicity, decreased cell viability and disrupted tight junctions between cells, while the knockdown of METTL3 reversed these results. Furthermore, METTL3 was involved in the inflammatory response of IPEC-J2 cells by activating the TLR2/NF-κB signaling pathway through regulating TLR2 m6A levels. In conclusion, METTL3 overexpression triggered the TLR2/NF-κB signaling pathway and promoted CPB2-induced inflammatory responses in IPEC-J2 cells. These findings may provide a new strategy for the prevention and treatment of diarrhea caused by .

摘要

β2 (CPB2) 毒素是 产生的主要致病毒素之一，可导致动物和人类的肠道疾病。N6-甲基腺苷（m6A）修饰是真核生物疾病过程中最常见的可逆修饰。甲基转移酶样 3（METTL3）调节动物细菌感染引起的免疫和炎症反应。然而，METTL3 参与 CPB2 处理的猪肠上皮细胞系-J2（IPEC-J2）的情况尚不清楚。在本研究中，我们使用甲基化 RNA 免疫沉淀-定量聚合酶链反应、Western blot 和免疫荧光测定来确定 METTL3 在 CPB2 暴露的 IPEC-J2 细胞中的作用。研究结果表明，CPB2 处理的 IPEC-J2 细胞中 m6A 和 METTL3 水平增加。功能上，METTL3 过表达促进了炎症因子的释放，增加了细胞毒性，降低了细胞活力，并破坏了细胞间的紧密连接，而 METTL3 的敲低则逆转了这些结果。此外，METTL3 通过调节 TLR2 m6A 水平，参与 IPEC-J2 细胞的 TLR2/NF-κB 信号通路的炎症反应。总之，METTL3 的过表达触发了 TLR2/NF-κB 信号通路，并促进了 IPEC-J2 细胞中 CPB2 诱导的炎症反应。这些发现可能为预防和治疗 引起的腹泻提供新的策略。

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