Dong Yachao, Xia Yandong, Yin Jie, Zhou Diao, Sang Yidan, Yan Sufeng, Liu Qingshu, Li Yaqi, Wang Leli, Zhao Ying, Chen Cang, Huang Qiuyun, Wang Ying, Abbasi Muhammad Nazeer, Yang Huansheng, Wang Chuni, Li Jianzhong, Tu Qiang, Yin Jia
Hunan Provincial Key Laboratory of Animal Intestinal Function and Regulation, Hunan International Joint Laboratory of Animal Intestinal Ecology and Health, The National and Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha, China.
CAS Key Laboratory of Quantitative Engineering Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences (CAS), Shenzhen, China.
Front Microbiol. 2022 Feb 21;12:761189. doi: 10.3389/fmicb.2021.761189. eCollection 2021.
Weaning of piglets could increase the risk of infecting with Gram-negative pathogens, which can further bring about a wide array of virulence factors including the endotoxin lipopolysaccharide (LPS). It is in common practice that the use of antibiotics has been restricted in animal husbandry. Alkaline phosphatase (AKP) plays an important role in the detoxification and anti-inflammatory effects of LPS. This study investigated the protective effects of AKP on intestinal epithelial cells during inflammation. Site-directed mutagenesis was performed to modulate the AKP activity. The enzyme activity tests showed that the activity of the DelSigDG-DN mutants in was nearly 1,600 times higher than that of the wild-type AKP. In this study, an LPS-induced inflammation model using IPEC-J2 cells was established. The mRNA expression of interleukin-(IL-) 6, IL-8, and tumor necrosis factor-α (TNF-α) were extremely significantly downregulated, and that of ASC amino acid transporter 2 (ASCT-2), zonula occludens protein-1 (ZO-1), and occludin-3 (CLDN-3) were significantly upregulated by the DelSigDG-DN mutant compared with LPS treatment. This concludes the anti-inflammatory role of AKP on epithelial membrane, and we are hopeful that this research could achieve a sustainable development for the pig industry.
仔猪断奶会增加感染革兰氏阴性病原体的风险，这进而会引发包括内毒素脂多糖（LPS）在内的一系列毒力因子。在畜牧业中，限制使用抗生素已成为普遍做法。碱性磷酸酶（AKP）在LPS的解毒和抗炎作用中发挥重要作用。本研究调查了AKP在炎症期间对肠上皮细胞的保护作用。进行了定点诱变以调节AKP活性。酶活性测试表明，DelSigDG-DN突变体的活性比野生型AKP高近1600倍。在本研究中，建立了使用IPEC-J2细胞的LPS诱导炎症模型。与LPS处理相比，DelSigDG-DN突变体使白细胞介素-（IL-）6、IL-8和肿瘤坏死因子-α（TNF-α）的mRNA表达极度显著下调，而ASC氨基酸转运体2（ASCT-2）、紧密连接蛋白-1（ZO-1）和闭合蛋白-3（CLDN-3）的mRNA表达显著上调。这总结了AKP在上皮细胞膜上的抗炎作用，我们希望这项研究能够实现养猪业的可持续发展。
