Arginine and Its Metabolites Attenuate Heat Stress-induced Apoptosis of Immature Boar Sertoli Cells
The aim of this study was to analyze the regulatory role of arginine and its metabolites in the induction of apoptosis in piglet sertoli cells under heat stress conditions.In this study,253-week-old healthy male piglets were selected,testes were collected and sertoli cells were isolated,which were randomly divided into 3 groups with 3 replicates.The cultured sertoli cells were treated at 44 ℃ for 30 min as a heat stress model.Mass spectrometry multiple reaction monitoring(MRM)was used to analyze the metabolites of amino acid.The apoptosis rate of cells was detected by flow cytometry.Western blotting was used to detect the expression of iNOS,BAX,FAS,ARG1,ODC and SSAT.Quantitative real-time PCR was used to detect the mRNA levels of Arg1,Arg2 and Odc.The contents of spermine were analyzed by the kit.The results showed that heat stress significantly decreased the levels of arginine(P＜0.01)and citrulline(P＜0.05),increased the levels of putrine(P＜0.01)and ornithine(P＜0.01),but did not affect the level of spermine(P＞0.05).Heat stress also significantly increased the mRNA expression of Inos(P＜0.01),Argl(P＜0.01),Arg2(P＜0.01)and Odc(P＜0.05)and the content of NO(P＜0.01),leading to an increase in the apoptosis rate(P＜0.01).Addition of 0.05 mmol·L-1 exogenous arginine increased the viability of SCs(P＜0.05)and arginine content(P＜0.01)under heat stress,and decreased BAX(P＜0.05)and FAS(P＜0.05)protein levels and relative apoptosis rate(P＜0.01),significantly reduced the protein level of iNOS(P＜0.01)and the content of NO(P＜0.01),increased the content of spermine(P＜0.01),and alleviated the apoptosis of SCs under heat stress(P＜0.01).Similar results were obtained with the addi-tion of exogenous spermine.These results indicate that heat treatment induces apoptosis of SCs by enhancing Arg-NO metabolic pathway.Addition of exogenous arginine and spermine enhances Arg-spermine metabolism and reduces NO production,thereby inhibiting heat stress-induced apoptosis of SCs.
万方数据
维普
