Paper Title
PROTECTIVE MECHANISMS OF A20 ON TESTOSTERONE SYNTHESIS IN LEYDIG CELLS UNDER AN AGING INFLAMMATORY ENVIRONMENT

Abstract
Abstract - Background: Aging-related inflammation is linked to reduced testosterone levels in males. A20, an important anti-inflammatory factor, is involved in inflammatory and age-related diseases. However, the association between A20 and testosterone production during aging has not been investigated. Methods: Bioinformatics analysis was conducted on rat testes data from the GEO dataset. Mouse testes were used for validation. Leydig cell lines were exposed to TNF-α to simulate an aging inflammatory environment. Lentiviral transduction manipulated A20 expression in these cell lines. Transcriptomic sequencing revealed differentially expressed genes in A20-overexpressing cells. Apoptosis, testosterone synthesis, and molecule expressions were assessed in testes and cells. Results: Bioinformatics analysis and validation experiments detected heightened inflammatory signaling and elevated A20 expression in aging rat and mouse testis. A20 knockdown aggravated TNF-induced testosterone synthesis inhibition and apoptosis in cells, while A20 over expression reversed this effect. Transcriptomic analysis found altered P38MAPK pathway with A20 over expression. A20 knockdown enhanced TNF-induced P38MAPK signaling, whereas its over expression attenuated it.A20 is involved in the regulation of testosterone synthesis by up regulating CEBPB expression. Conclusions: A20 inhibits apoptosis by blocking P38MAPK signaling and promotes the expression of CEBPB, consequently upregulating testosterone synthesis. Keywords - TNFAIP3/A20, Inflammation, CEBPB, Leydig Cell, Testosterone Synthesis