De León-Ramírez Yeimy Mar, Nicolás-Toledo Leticia, Pérez-Sánchez Eliut, Arroyo-Helguera Omar
Laboratorio de Biomedicina y Salud Pública, Instituto de Salud Pública, Universidad Veracruzana, Av. Luís Castelazo Ayala S/N, Col. Industrial Animas, Xalapa C.P. 91190, Veracruz, Mexico.
Centro Tlaxcala de Biología de la Conducta, Universidad Autónoma de Tlaxcala, Km 1.5 Carretera Tlaxcala-Puebla S/N, La Loma Xicoténcatl, Tlaxcala C.P. 90070, Tlaxcala, Mexico.
Biology (Basel). 2025 Jun 12;14(6):690. doi: 10.3390/biology14060690.
Stressor stimuli induce oxidative stress and functional abnormalities in sperm, which are linked to a reduced sperm quality and male infertility. Furthermore, oxidative stress can trigger cell death. However, the impact of stressor stimulation on testicles and epididymal sperms and apoptosis has not been explored. This study analyzes the expression of extrinsic and intrinsic apoptotic markers in the testicle and epididymis of rats exposed to chronic variable stress (CVS). We used male Wistar rats divided into two groups: the control group was kept undisrupted, and the stress group was stressed daily using a CVS model for four weeks, except for the weekends (from postnatal days 51 to 81). After the last week, the rats were sacrificed, and complete testicles and epididymal sperm were used to measure oxidative stress and the total antioxidant status by colorimetric methods. The expressions of PPAR-γ, p53, Bax, and Bcl-2 markers at the mRNA level were determined by real-time PCR, and the p-Akt, AP-2α, PPAR-γ, C/EBP-β and FAS protein levels were detected by immunoblot. The results showed low levels of p-Akt and AP-2α proteins and high levels of FAS, PPAR-γ, and C/EBP-β in the testicle and epididymis of rats exposed to CVS. At the mRNA level, we observed the upregulation of PPAR-γ, p53, p21, HIF-α, and Bax expressions in the epididymis of rats exposed to CVS, consistent with the significant caspase-3 activity observed in both the epididymis and testicles in the CVS group. In conclusion, CVS damage triggers the induction of apoptosis markers by intrinsic (PPAR-γ, p53, p21, HIF-α, and Bax) and extrinsic (p-Akt, AP-2α, and FAS) caspase-3-dependent pathways in complete extracts of both the testicles and epididymis. This study supports the view that stressor stimuli could be involved in the infertility process.
应激源刺激会诱导精子产生氧化应激和功能异常，这与精子质量下降和男性不育有关。此外，氧化应激会引发细胞死亡。然而，应激源刺激对睾丸和附睾精子以及细胞凋亡的影响尚未得到研究。本研究分析了暴露于慢性可变应激（CVS）的大鼠睾丸和附睾中外源性和内源性凋亡标志物的表达。我们将雄性Wistar大鼠分为两组：对照组保持不受干扰，应激组除周末外（从出生后第51天至81天）每天使用CVS模型进行应激处理，持续四周。在最后一周结束后，处死大鼠，使用完整的睾丸和附睾精子通过比色法测量氧化应激和总抗氧化状态。通过实时PCR测定PPAR-γ、p53、Bax和Bcl-2标志物在mRNA水平的表达，并通过免疫印迹检测p-Akt、AP-2α、PPAR-γ、C/EBP-β和FAS蛋白水平。结果显示，暴露于CVS的大鼠睾丸和附睾中p-Akt和AP-2α蛋白水平较低，而FAS、PPAR-γ和C/EBP-β水平较高。在mRNA水平上，我们观察到暴露于CVS的大鼠附睾中PPAR-γ、p53、p21、HIF-α和Bax表达上调，这与CVS组附睾和睾丸中观察到的显著的caspase-3活性一致。总之，CVS损伤通过内源性（PPAR-γ、p53、p21、HIF-α和Bax）和外源性（p-Akt、AP-2α和FAS）caspase-3依赖性途径在睾丸和附睾的完整提取物中触发凋亡标志物的诱导。本研究支持应激源刺激可能参与不育过程这一观点。
