Multi-omics analysis of chlorogenic acid in alleviating oxidative damage from heat stress in prepubertal boar testes

Background: Heat stress (HS) can impair boar testicular function, leading to reproductive issues. While chlorogenic acid (CGA) has shown function in mitigating HS-induced damage in various livestock and poultry species. Prepuberty is an important stage of testicular development in boars after birth. However, the protective effect of CGA on testicular HS injury during prepuberty boars and the underlying mechanisms are still not fully understood. Results: In this study, we established an in vivo HS model using prepubertal boars. The results showed that CGA mitigated the HS-induced reduction in testicular tissue antioxidant capacity, decline in Sertoli cell count, and decrease in sperm density post sexual maturation. Utilizing 1H-NMR, RNA-seq and TMT proteomics, we observed alterations in glutathione and other compounds levels in the testicular tissue due to HS in prepubertal boars. Furthermore, the differentially expressed genes influenced by CGA and HS were predominantly associated with the glutathione metabolism pathway. The combined analysis of transcriptomics and proteomics showed that only BLVRA was affected by both HS and CGA when the mRNA and protein levels of a gene showed differential expression with the same trend. Further in vitro studies confirmed that CGA modulated GPX3 expression via BLVRA, impacted GPx activity, and attenuated HS-induced ROS accumulation. Conclusions: Our findings demonstrate that HS in prepubertal boars can impair their ability to produce sperm after sexual maturity. CGA inhibits HS-induced oxidative stress injury in Sertoli cells by restoring GPx activity through BLVRA. This study contributes to elucidating the mechanisms underlying HS effects on prepubertal boar testicular development using multi-omics approaches, thereby laying a foundation for the potential utilization of CGA in swine production.