A quantitative proteomic analyses of primary myocardial cell injury induced by heat stress in chicken embryo

Highlights

• 245 differentially expressed proteins (DEPs) were identified and many were related to metabolism, oxidative stress, oxidative phosphorylation and apoptosis.

• The electron transfer and REDOX reaction in the respiratory chain were affected.

Abstract

In this study, the model of heat stress was constructed in primary chick embryonic myocardial cells at 42 °C for 4 h. Proteome analysis using DIA identified 245 differentially expressed proteins (DEPs) (Q-value <0.05, fold change >1.5), of which 63 proteins were up-regulated and 182 proteins were down-regulated. Many were related to metabolism, oxidative stress, oxidative phosphorylation and apoptosis. Gene Ontology (GO) analysis showed that many DEPs under heat stress were involved in regulating metabolites and energy, cellular respiration, catalytic activity and stimulation. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that DEPs were enriched in metabolic pathways, oxidative phosphorylation, citrate cycle (TCA cycle), cardiac muscle contraction, and carbon metabolism. The results could help understanding of the effect of heat stress on myocardial cells and even the heart and possible action mechanism at the protein level.

Introduction

Poultry are homeotherms with particularly sensitive to high temperatures, because they are covered with feathers and with under-developed sweat glands. Therefore, under high temperature, poultry experience stress more than other species (Park et al., 2013). The significant effects of heat stress on poultry include changes in the respiratory metabolism, energy demand and blood circulation (Zaboli et al., 2017), which aggravate the burden of the heart (Chang et al., 2003).

In response to the challenge of heat stress, the various defense and repair system of poultry were activated, and then bring a series of biochemical and physiological changes, among which the most typical is the antioxidant system (Slimen et al., 2016; Gonzalez-Rivas et al., 2020; Goel, 2021; Vandana et al., 2021). The various systems and organs of the body maybe have different changes under heat stress. The heart is an important organ related with energy supply. Both contraction and diastole of the heart are energy expenditure process, and the heart needs a large amount of energy in the form of ATP to sustain its systolic and diastolic function (Bertero and Maack, 2018), and understanding the response mechanisms of heart to heat stress should aid in taking measures to relieve heat stress from the point of view of energy metabolism. Previous studies have shown that myocardial cells are the early target of heat stress (Wu et al., 2015), persistent heat stress caused myocardial damage, heart failure, sudden arrest, and even sudden death in animals (Syafwan et al., 2011). There have been reported that metabolic remodeling in failing hearts is characterized by defects in energy production and changes in metabolic pathways, and that involve the regulatory functions of relevant basic cells (Mcgarrah et al., 2018). Studies of the metabolic changes of heart failure have focused on the metabolism of fatty acids and glucose (Doenst et al., 2013). The mechanism of cellular self-protection against stress is complex, involving regulation of many genes and proteins (Tang et al., 2015). However, at present, studies on heat stress in poultry have mainly focused on physiological changes, production performance and heat shock proteins, while there were less on proteomic studies (Liu et al., 2020). Thus, it is necessary to understand the molecular mechanisms that operate in response to heat stress at the protein level.

Data-independent acquisition (DIA) is a high sensitivity and resolution two-stage mass spectrometry (MS) quantitative proteomics technique, which have achieved a continuous, cyclic and full-coverage collection to all fragment ion signals (Doerr, 2015). The advantages of DIA were high repeatability and high throughput, which are conducive to large-scale research on gene and cell function at the protein level (Shruthi et al., 2016; Monti et al., 2019), and has been widely used in biological and clinical studies (Musa et al., 2018; Floriou-Servou et al., 2018).

Based on the above, it was hypothesized that the cardiomyocyte proteome might be undergo some interesting changes under heat stress, and which maybe also help explain cardiomyocyte damage or death caused by heat stress. Therefore, in the present study, we analyzed proteome changes in primary chick embryonic myocardial cells under heat stress using DIA technology to explore the mechanism of heat stress at the protein level.