Quantitative proteomics reveals manganese alleviates heat stress of broiler myocardial cells via regulating nucleic acid metabolism

Abstract

Abstract Heat stress threatens severely cardiac function by caused myocardial injury in poultry. Our prebious study have showed that Manganese (Mn) has a beneficial effect on heat-stress resistance of broiler. Therefore, we tried to confirm the alleviation mechanism through proteomic analysis after heat stress exposure to primary broiler myocardial cells pretreated with Mn. The experiment was divided into four groups: CON group (37°C, cells without any treatment), HS group (43°C, cells treatment with heat stress for 4 h), HS + MnCl2 group (cells treated with 20 µM MnCl2 before heat stress) and HS + Mn-AA group (cells treated with 20 µM Mn compound amino acid complex before heat stress). Proteome analysis using DIA identified 300 differentially expressed proteins (DEPs) between CON group and HS group, 93 and 121 DEPs were identified in inorganic manganese treatment group and organic manganese treatment group respectively, in addition, there were 53 DEPs identified between inorganic and organic manganese group. Gene Ontology (GO) analysis showed that DEPs were mainly involved in binding, catalytic activity, response to stimulus and metabolic process. DEPs of manganese pertreatment involved in a variety of biological regulatory pathways, and significantly influenced protein processing and repair in endoplasmic reticulum, apoptosis and DNA replication and repair. These all seems to imply that manganese may help to resist cell damage induced by heat stress by regulating key node proteins. These findings contribute to a better understanding of the effects of manganese on overall protein changes during heat-stress and the possible mechanisms, as well as how to better use manganese to protect heart function in high temperature.