Identification and Analysis of Liver Differentially Expressed Proteins in Shaoxing Duck (Anas platyrhynchos) Under Acute Heat Stress Based on iTRAQ
WANG De-Qian1, DONG Jie1, XU Ning-Ying2, HUANG Min-Jie1,*
1 Institute of Animal Husbandry and Veterinary Science, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China; 2 College of Animal Science, Zhejiang University, Hangzhou 310058, China
Abstract：In recent years, with the increasing intensification of laying duck (Anas platyrhynchos) cultivation, the effect of heat stress caused by constant high temperature in summer poses a serious challenge to safety production. To elucidate the effect of heat stress on hepatic protein expression and to explore the function of differential protein genes in laying duck, in this study, Shaoxing ducks at the peak laying period (245-day old) were selected for heat stress treatment and liver tissues were collected. A total of 98 differentially expressed proteins (DEPs) were identified by isobaric tags for relative and absolute quantification (iTRAQ) and LC-MS/MS, with significantly upregulation of heat shock protein 60 (HSP60)(P<0.01). KEGG analysis showed that DEPs enriched in 15 metabolic pathways after heat stress. Glycolysis and gluconeogenesis, tricarboxylic acid cycle and pyruvate may be related to the effects of heat stress. GO analysis showed that DEPs were involved in translation extension, glucose catabolism process, glycolysis. HSP60-siRNA was transfected into duck hepatocytes after screening and optimization. Compared with negative control (NC) group, HSP60 gene expression was decreased by 75.69% (P<0.01), cytochrome coxidase gene (CCO)(P<0.01), and stress-activated protein kinase gene (SAPK) were significantly increased (P<0.01), protein kinase gene (PKB) was significantly decreased (P<0.01). There was no significant difference in the expression of tumor necrosis factor-related apoptosis inducing receptors gene (TRAIL-Rs)(P>0.05). The results showed that HSP60 was significantly increased in the liver of laying ducks under acute heat stress and inhibited apoptosis by regulating the expression of key genes of apoptotic signaling pathways. It is helpful to prevent and control the occurrence of heat stress by preliminarily exploring the mechanism of heat stress occurrence in laying ducks.
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