玉米赤霉烯酮诱导山羊子宫内膜基质细胞凋亡的研究

doi:10.11843/j.issn.0366-6964.2021.02.026

摘要/Abstract

摘要： 玉米赤霉烯酮（zearalenone，ZEN）是由镰刀菌产生的一种霉菌毒素，具有细胞毒性和雌激素活性，会对生殖系统、泌尿系统和消化系统等产生严重毒性影响。目前，ZEN对山羊子宫内膜基质细胞（endometrial stromal cells，ESCs）凋亡的潜在作用仍不清楚。本研究通过体外培养山羊永生化ESCs，采用CCK-8和流式细胞术检测ZEN对山羊ESCs增殖和凋亡的影响；应用Real-time PCR和Western blot技术检测内质网应激（endoplasmic reticulum stress，ERS）和凋亡关键基因的变化，探讨ZEN是否通过ERS影响山羊ESCs的增殖与凋亡。结果显示，ZEN抑制了山羊ESCs的增殖过程并呈剂量依赖性关系。随着ZEN剂量的增加，山羊ESCs的凋亡比例显著上升。ZEN剂量依赖性地上调了Caspase-8、Caspase-9及Bax/Bcl-2 mRNA表达水平。ZEN显著上调了ERS标志分子GRP78和CHOP mRNA和蛋白的表达水平，同时，ERS关键分子ATF6、PERK和IRE1 mRNA表达水平显著增加，并且ZEN对IRE1通路的影响更为显著。添加ERS特异性抑制剂4-PBA和IRE1特异性抑制剂Irestatin 9389均可显著降低ZEN对ESCs增殖的抑制作用。综上表明，ZEN可以通过ERS通路调控山羊ESCs的凋亡进程。

关键词: 玉米赤霉烯酮, 子宫内膜基质细胞, 细胞凋亡, 内质网应激, 山羊

Abstract: Zearalenone (ZEN) is a fungal toxin with cytotoxic and estrogenic activity which is produced by Fusarium, ZEN has serious toxic impacts on the reproductive system, urinary system and digestive system. At present, the potential mechanism of ZEN on apoptosis of goat endometrial stromal cells (ESCs) remains unclear. In this study, we studied the effect of ZEN on the proliferation and apoptosis of goat ESCs by CCK-8 and flow cytometry in vitro. To investigate whether ZEN affected the proliferation and apoptosis of goat ESCs through endoplasmic reticulum stress (ERS), Real-time PCR and Western blot were used to detect the gene expression levels of ERS and apoptosis. The results showed that ZEN inhibited the proliferation of goat ESCs in a dose-dependent manner. With the increase of ZEN dose, the apoptosis rate of goat ESCs increased significantly. ZEN dose-dependent up-regulation of Caspase-8, Caspase-9 and Bax/Bcl-2 mRNA expression levels. ZEN could significantly up-regulate the expression levels of GRP78 and CHOP mRNA and protein. Meanwhile, the expression levels of ATF6, PERK and IRE1 mRNA were drastically increased. In contrast, the IRE1 pathway could be significantly affected by ZEN. Addition of ERS specific inhibitor 4-PBA or IRE1 specific inhibitor Irestatin 9389 significantly reduced the inhibitory effect of ZEN on ESCs proliferation. These results indicated that ZEN could regulate the apoptotic process of ESCs in goats through the ERS pathway.

Key words: zearalenone, endometrial stromal cells, apoptosis, endoplasmic reticulum stress, goat

中图分类号:

S856.9

王宗捷, 张瑞雪, 刘守勤, 靳亚平, 林鹏飞. 玉米赤霉烯酮诱导山羊子宫内膜基质细胞凋亡的研究[J]. 畜牧兽医学报, 2021, 52(2): 535-542.

WANG Zongjie, ZHANG Ruixue, LIU Shouqin, JIN Yaping, LIN Pengfei. The Apoptotic Effect of Zearalenone on Goat Endometrial Stromal Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(2): 535-542.

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