急性热应激通过降低牛磺酸水平影响猪睾丸支持细胞的功能

doi:10.11843/j.issn.0366-6964.2025.04.026

摘要/Abstract

摘要：

旨在揭示猪睾丸支持细胞对急性热应激的代谢应答。本研究于急性热应激前(Control组)、急性热应激(43 ℃，0.5 h)刚结束(HS0.5组)和急性热应激后恢复36 h(HS0.5-R36组)收集猪睾丸支持细胞(每组3个重复)进行液相质谱(LC-MS/MS)检测，ELISA验证差异代谢物。通过CCK-8、EdU和Annexin V FITC/PI试剂盒检测关键差异代谢物对猪睾丸支持细胞的细胞活力、细胞增殖和细胞凋亡效应。利用ROS、RH123和Mito Tracker染色检测细胞ROS水平、线粒体分布与线粒体膜电位情况。结果，急性热应激前后(HS0.5 vs. Control)、恢复36 h相对急性热应激刚结束(HS0.5-R36 vs. HS0.5)、恢复36 h相对急性热应激前(HS0.5-R36 vs. Control)分别诱导33个、57个和115个显著差异(P < 0.05；VIP>1.00)二级代谢物。通路富集分析显示，显著差异代谢物主要参与牛磺酸和亚牛磺酸代谢等通路。ELISA验证结果显示，急性热应激显著降低牛磺酸和γ-氨基丁酸水平，与代谢组检测的变化趋势一致。HS0.5-R36 vs. Control牛磺酸下降约5.7 μmol·L^-1，而添加5.7 μmol·L^-1牛磺酸处理猪睾丸支持细胞48 h显著增加细胞存活率、促进增殖、抑制凋亡和增强线粒体功能(P < 0.05)。急性热应激显著改变猪睾丸支持细胞的代谢，显著降低的牛磺酸部分介导了急性热应激对猪睾丸支持细胞的功能损伤。本研究结果为猪睾丸支持细胞热应激损伤拯救提供了新的参考。

关键词: 猪, 睾丸支持细胞, 急性热应激, 代谢组学, 牛磺酸

Abstract:

To investigate the metabolic response to acute heat stress of porcine sertoli cells, porcine sertoli cells were collected under 3 conditions: before acute heat stress (control group), immediately after acute heat stress (43℃, 0.5 h; HS0.5 group), and after a 36 hour recovery post-acute heat stress (HS0.5-R36 group), with 3 biological replicates per group. Metabolic profiling was detected using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Differential metabolites were validated via ELISA. The effects of key differential metabolite on cell viability, proliferation, and apoptosis were assessed using CCK-8, EdU, and Annexin V FITC/PI kits. Level of reactive oxygen species (ROS), mitochondrion distribution, and mitochondrion membrane potential were evaluated using ROS, RH123 and Mito Tracker staining. The results showed that 33, 57 and 115 significant differential secondary metabolites (P < 0.05; VIP>1.00) were identified respectively in HS0.5 vs. Control, HS0.5-R36 vs. HS0.5 and HS0.5-R36 vs. Control comparisons. The pathway analysis showed that significant differential metabolites were mainly enriched in taurine and hypotaurine metabolism and other pathways. ELISA verified that acute heat stress significantly decreased levels of taurine and gamma-aminobutyric acid, which were consistent to metabolomic results. Taurine level was significantly decreased around 5.7 μmol·L^-1 in HS0.5-R36 group as compared to control group. Treatment of porcine sertoli cells using taurine of 5.7 μmol·L^-1 for 48 h significantly increased cell viability, promoted proliferation, suppressed apoptosis and enhanced mitochondrial function (P < 0.05). These results suggest that acute heat stress could significantly alter metabolism, and the differential metabolite taurine partially mediate the damage exerted by acute heat stress on porcine sertoli cells. These results provide new insights for rescuing damage caused by acute heat stress on porcine sertoli cells.

Key words: porcine, sertoli cell, acute heat stress, metabolomics, taurine

中图分类号:

S828.3

王昕昕, 刘小英, 王宜, 王芳, 赵晗, 杜志强, 杨彩侠. 急性热应激通过降低牛磺酸水平影响猪睾丸支持细胞的功能[J]. 畜牧兽医学报, 2025, 56(4): 1779-1790.

WANG Xinxin, LIU Xiaoying, WANG Yi, WANG Fang, ZHAO Han, DU Zhiqiang, YANG Caixia. Acute Heat Stress Affects the Functions of Porcine Sertoli Cells via Decreasing Taurine Level[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(4): 1779-1790.

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