Methionine deficien y inhibited pyroptosis in primary hepatocytes of grass carp(Ctenopharyngodon idella):possibly via activating the ROS-AMPK-autophagy axis  

作　　者：Yuanlin He Pei Wu Weidan Jiang Yang Liu Xiaowan Jin Hongmei Ren Ruinan Zhang Xiaoqiu Zhou Lin Feng 

机构地区：[1]Animal Nutrition Institute,Sichuan Agricultural University,Chengdu 611130,Sichuan,China [2]Fish Nutrition and Safety Production,University Key Laboratory of Sichuan Province,Sichuan Agricultural University,Chengdu 611130,Sichuan,China [3]Key Laboratory of Animal Disease-Resistant Nutrition and Feed,Ministry of Agriculture and Rural Affairs,Chengdu,China [4]Key Laboratory of Animal Disease-Resistant Nutrition,Chengdu,Sichuan,China

出　　处：《Journal of Animal Science and Biotechnology》2024年第6期2607-2624,共18页畜牧与生物技术杂志(英文版)

基　　金：financially supported by National Natural Science Foundation of China(32072985,32273144);the earmarked fund for CARS(CARS-45);National Key R&D Program of China(2019YFD0900200)。

摘　　要：Background Methionine(Met)is the only sulfur-containing amino acid among animal essential amino acids,and methionine deficiency(MD)causes tissue damage and cell death in animals.The common modes of cell death include apoptosis,autophagy,pyroptosis,necroptosis.Howeve r,the studies about the major modes of cell death caused by MD have not been reported,which worth further study.Methods Primary hepatocytes from grass carp were isolated and treated with different doses of Met(0,0.5,1,1.5,2,2.5 mmol/L)to examine the expression of apoptosis,pyroptosis,autophagy and necroptosis-related proteins.Based on this,we subsequently modeled pyroptosis using lipopolysaccharides and nigericin sodium salt,then autophagy inhibitors chloroquine(CQ),AMP-activated protein kinase(AMPK)inhibitors compound C(CC)and reactive oxygen species(ROS)scavengers N-acetyl-L-cysteine(NAC)were further used to examine the expression of proteins related to pyroptosis,autophagy and AMPK pathway in MD-treated cells respectively.Results MD up-regulated B-cell lymphoma protein 2(Bax),microtubule-associated protein 1 light chain 3Ⅱ(LC3Ⅱ),and down-regulated the protein expression levels of B-cell lymphoma-2(Bcl-2),sequestosome 1(p62),cleavedcaspase-1,cleaved-interleukin(IL)-1βand receptor-interacting protein kinase(RIP)1 in hepatocytes,while it did not significantly affect RIPS.In addition,MD significantly increased the protein expression of liver kinase B1(LKB1),p-AMPK,and Unc-51-like kinase 1(ULK1)without significant effect on p-target of rapamycin.Subsequently,the use of CQ increased the protein expression of NOD-like receptor thermal protein domain associated protein 3(NLRP3),cleaved-caspase-1,and cleaved-IL-1βinhibited by MD;the use of CC significantly decreased the protein expression of MD-induced LC3Ⅱand increased the protein expression of MD-suppressed p62;then the use of NAC decreased the MD-induced p-AMPK protein expression.Conclusion MD promoted autophagy and apoptosis,but inhibited pyroptosis and necroptosis.MD inhibited pyroptosis may be

关 键 词：AUTOPHAGY METHIONINE Primary hepatocytes PYROPTOSIS 

分 类 号：S917.4[农业科学—水产科学]