α-Linolenic acid promotes testosterone synthesis by improving mitochondrial function in primary rooster Leydig cells

• Published in: Theriogenology 2025-01, Vol.232, p.9-19
• Main Authors: Chang, Xuerui, Li, Danyang, Guo, Yong, Sheng, Xihui, Wang, Xiangguo, Xing, Kai, Xiao, Longfei, Lv, Xueze, Long, Cheng, Qi, Xiaolong
• Format: Article
• Language: English
• Subjects: alpha-Linolenic Acid - pharmacology; Animals; Antioxidant capability; Apoptosis - drug effects; Cell Survival - drug effects; Cells, Cultured; Chickens; Leydig cells; Leydig Cells - drug effects; Leydig Cells - metabolism; Male; Membrane Potential, Mitochondrial - drug effects; Mitochondria - drug effects; Mitochondria - metabolism; Mitochondrial biogenesis; Rooster; Testosterone - biosynthesis; Testosterone - metabolism; Testosterone production; α-Linolenic acid
• ISSN: 0093-691X; 1879-3231; 1879-3231
• DOI: 10.1016/j.theriogenology.2024.10.026

The present study aimed to investigate the direct effects of α-Linolenic acid (ALA) on the in vitro production of testosterone and the expression of key enzymes and proteins related to steroidogenesis in Leydig cells of roosters. Purified primary Leydig cells isolated from 65-week-old roosters were purified and treated with different concentrations of ALA treatments: (0 μm/L [control], solvent control group (DMSO), 20 μM/L, 40 μM/L, and 80 μM/L) and cell counting-8 (CCK-8) for cell viability assay, Enzyme-linked immunosorbent assay (ELISA) kit for the determination of testosterone in cell supernatants, quantitative (real-time) PCR, and analysis of activities of antioxidants catalase (CAT), superoxide dismutase (SOD) and malondialdehyde (MDA), evaluation of mitochondrial membrane potential, pro- and anti-apoptotic proteins/genes Bcl-2, Bcl-2-associated X protein (Bax), apoptosis-inducing factor (AIF) were done respectively. Our results showed that ALA significantly increased testosterone secretion in primary rooster Leydig cells (P