Acta Veterinaria et Zootechnica Sinica ›› 2025, Vol. 56 ›› Issue (11): 5575-5587.doi: 10.11843/j.issn.0366-6964.2025.11.018

• Animal Biotechnology and Reproduction • Previous Articles     Next Articles

The Impact of Gossypol on Spermatogenic Capacity of the Testis and Apoptosis of Sertoli Cells

CHEN Qian1,2,3(), TONG Jiang1,2,3, PAN Yuheng1,2,3, MA Jianfeng1,2,3, SHI Yuqian1,2,3, CHEN Siyu1,2,3, LI Jiaxin1,2,3, ZHOU Ting4, LIU Chendong4, ZHU Li1,2,3, SHEN Linyuan1,2,3,*(), GAN Mailin1,2,3,*()   

  1. 1. National Key Laboratory of Pig and Poultry Seed Industry, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, China
    2. Key Laboratory of Livestock and Poultry Biomics of Ministry of Agriculture and Rural Affairs, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, China
    3. Sichuan Provincial Key Laboratory of Exploration and Innovative Utilization of Livestock and Poultry Genetic Resources, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, China
    4. Sichuan Litian Animal Husbandry Co., Ltd. Nutrition Department, Dazhou 635000, China
  • Received:2025-04-09 Online:2025-11-23 Published:2025-11-27
  • Contact: SHEN Linyuan, GAN Mailin E-mail:chenqian1@stu.sicau.edu.cn;shenlinyuan@sicau.edu.cn;ganmailin@sicau.edu.cn

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Abstract:

The aim of this study was to explore the reproductive toxicity mechanism of gossypol-induced testicular tissue injury and TM4 cells(mouse testicular sertoli cells) apoptosis in mice. A gossypol intragastric administration model in male ICR(improved castle road) mice (0, 30 (mg·kg) ·d-1) for 30 consecutive days) and a gossypol exposure model in TM4 cells (0, 15, 20, 25 μmol·L-1) were established. Subsequently, the sperm quality of mice was assessed by an automatic sperm analysis system; the histopathological alterations of testicular tissue were observed via HE staining; and the expression levels of apoptosis, autophagy, ferroptosis, tight junction, and spermatogenesis-related genes in testicular tissue, as well as apoptosis-related genes in TM4 cells, were detected using real-time fluorescence quantitative PCR, and the expression of apoptosis-related proteins in testicular tissue was detected by Western blot. The findings revealed that gossypol significantly decreased the weight gain rate, sperm motility, and sperm density of mice (P < 0.01), and elicited histopathological changes in testicular tissue (such as atrophy of seminiferous tubules, destruction of some sertoli cells, and reduction in sperm count). The expression trends of genes related to the three death modes of ferroptosis, autophagy, and apoptosis in testicular tissue all changed, and the expression trend of apoptosis-related genes changed most significantly. The expression of the apoptosis inhibitory gene Bcl-2 was significantly downregulated (P < 0.05), while the expressions of the apoptosis promoting genes Bax, Tnf-α, and the Bax/Bcl-2 ratio were upregulated (P < 0.01, P < 0.05, P < 0.01). The expressions of apoptosis proteins Bax, Tnf-α, and the Bax/Bcl-2 ratio in testicular tissue were significantly increased (P < 0.05). Additionally, the expressions of tight junction and spermatogenesis-related genes in testicular tissue were significantly downregulated. In the in vitro experiment with TM4 cells, exposure to different concentrations of gossypol significantly influenced the expression of apoptosis-related genes within TM4 cells, and the expressions of apoptosis promoting genes Bax, Tnf-α, and the Bax/Bcl-2 ratio were Extremely significantly upregulated (P < 0.01), which was consistent with the quantitative results of testicular tissue. In summary, the weakened spermatogenic ability of the testis in gossypol-exposed mice is closely related to the apoptosis of TM4 cells, and this apoptosis is associated with the increased expressions of Bax, Tnf-α, and Bax/Bcl-2.

Key words: gossypol, TM4 cells, RT-qPCR, apoptosis, Bax, Tnf-α, Bax/Bcl-2

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