Abstract
Variations in disease resistance among pig breeds have been extensively documented, with Sertoli cells (SCs) playing a pivotal role in spermatogenesis. Infections can induce oxidative stress, which can lead to damage to these cells. This study aimed to compare the levels of oxidative stress in SCs from Rongchang and Landrace pig breeds following LPS challenge. SCs were isolated, cultured, and stimulated with LPS to assess cell viability and markers of oxidative stress. Cell viability was evaluated along with oxidative stress markers such as reactive oxygen species (ROS), mitochondrial superoxide, malondialdehyde, and antioxidant enzymes. Mitochondrial function was assessed using JC-1 and Calcein AM probes. Transcriptomic analysis identified differentially expressed genes (DEGs), while ingenuity pathway analysis (IPA) explored enriched pathways. IL20RA, identified through transcriptomics, was validated using the siRNA knockdown technique. The results showed that Rongchang SCs exhibited lower levels of oxidative stress compared to Landrace SCs along with higher activity of antioxidant enzymes. IL20RA emerged as a key regulator since its knockdown affected mitochondrial superoxide production and catalase secretion. The findings suggest that Rongchang SCs possess superior antioxidant capacity, possibly due to the IL20RA-mediated protection of mitochondria, thereby providing insights into breed-specific resistance against oxidative stress and highlighting the role of IL20RA in maintaining stem cell function.