猪METTL3基因表达水平与DON诱导IPEC-J2细胞损伤的关系

doi:10.11843/j.issn.0366-6964.2022.03.003

摘要/Abstract

摘要： 旨在初步探究猪m⁶A甲基化酶METTL3基因表达水平与脱氧雪腐镰刀菌烯醇(deoxynivalenol,DON)诱导猪小肠上皮细胞(IPEC-J2)损伤的关系。本研究构建了稳定干扰METTL3基因表达水平的IPEC-J2细胞系,用1 μg·mL^-1 DON诱导METTL3干扰组和对照组猪肠上皮细胞48 h,通过实时荧光定量PCR检测凋亡相关基因、免疫相关基因、抗氧化酶相关基因表达水平,同时测定细胞增殖活性、细胞周期、细胞凋亡以及活性氧(ROS)水平。结果显示,与对照组相比,干扰组免疫相关基因TNF-α、IL-6、IL-12的表达水平产生了显著变化(P<0.05),凋亡相关基因Bax、Caspase3、Caspase9表达量均显著下降(P<0.05),抗氧化酶相关基因GPXs、CuZn-SOD、CAT表达量显著上升(P<0.05);细胞增殖活性显著提高,细胞总凋亡率显著降低(P<0.05),ROS水平极显著降低(P<0.01)。本研究结果表明,METTL3基因表达水平降低能一定程度地帮助IPEC-J2细胞抵抗因DON诱发的细胞凋亡和炎症。METTL3表达下调有利于缓解DON对于细胞损伤产生的一系列炎症反应,为今后细致研究RNA甲基化修饰在仔猪抵抗脱氧雪腐镰刀菌烯醇损害调控中的作用机制提供了理论基础。

关键词: 猪, METTL3, 脱氧雪腐镰刀菌烯醇

Abstract: This study aimed to investigate the relationship between the expression level of porcine m⁶A methylases METTL3 and deoxynivalenol (DON) induced injury of intestinal porcine epithelial cells (IPEC-J2). In this study, IPEC-J2 cell lines with METTL3 knockdown was successfully established. IPEC-J2 cells from the METTL3 interference group and control group were treated with 1 μg·mL^-1 DON for 48 h, respectively. Real-time quantitative PCR was used to detect the mRNA expression differences of apoptosis-related genes, immune-related genes and oxidase-related genes, and cell proliferation activity, cell cycle, cell apoptosis, and reactive oxygen species (ROS) levels were measured. The results showed that, compared with control group, the expression of immune-related genes TNF-α, IL-6 and IL-12 in the interference group changed significantly (P<0.05); The levels of apoptosis-related genes Bax, Caspase3 and Caspase9 were significantly decreased(P<0.05); The levels of oxidase-related genes GPXs, CuZn-SOD and CAT were significantly increased(P<0.05). The cell proliferation activity was significantly increased, and the total cell apoptosis rate was significantly decreased (P<0.05); The level of ROS was extremely significantly decreased (P<0.01). The results indicate that the down-regulation expression of METTL3 gene contributes to the enhancement of IPEC-J2 cells resisting apoptosis and inflammation induced by DON in a certain extent. The down-regulation expression of METTL3 gene is beneficial to alleviate a series of inflammatory responses and cell damage caused by DON, and will provide a theoretical basis for in-depth study of the mechanism of RNA methylation in the regulation of piglet resistance to DON damage in the future.

Key words: pigs, METTL3, deoxynivalenol

中图分类号:

S828.2

许写, 石东风, 吴嘉韵, 吴圣龙, 王海飞, 吴正常, 包文斌. 猪METTL3基因表达水平与DON诱导IPEC-J2细胞损伤的关系[J]. 畜牧兽医学报, 2022, 53(3): 688-699.

XU Xie, SHI Dongfeng, WU Jiayun, WU Shenglong, WANG Haifei, WU Zhengchang, BAO Wenbin. Relationship between Expression Level of Porcine METTL3 Gene and the Injury of IPEC-J2 Cells Induced by DON[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(3): 688-699.

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