研究论文 RESEARCH PAPER

姜酮缓解3-乙酰呕吐毒素诱导的猪肠上皮细胞损伤研究

  • 张同坤 ,
  • 贾海 ,
  • 季昀 ,
  • 杨鹰 ,
  • 武振龙
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  • 中国农业大学动物科学技术学院, 北京 100193
张同坤(1995-),男,安徽合肥人,硕士研究生,动物营养与饲料科学专业。E-mail:18225690807@163.com

收稿日期: 2022-04-13

  网络出版日期: 2022-10-17

基金资助

国家自然科学基金项目(32172749)

Study on Zingerone Alleviating Injury of Porcine Intestinal Epithelial Cells Induced by 3-Acetyldeoxynivalenol

  • ZHANG Tongkun ,
  • JIA Hai ,
  • JI Yun ,
  • YANG Ying ,
  • WU Zhenlong
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  • College of Animal Science and Technology, China Agricultural University, Beijing 100193, China

Received date: 2022-04-13

  Online published: 2022-10-17

摘要

本研究旨在探讨姜酮对3-乙酰呕吐毒素(3-Ac-DON)诱导的猪肠上皮细胞损伤的影响。选用IPEC-1细胞经16 μmol/L的3-Ac-DON和0、80、160和320 μg/mL的姜酮共处理24 h,选择合适的姜酮浓度,研究其对细胞凋亡、内质网应激相关蛋白和紧密连接蛋白表达的影响。结果表明: 1)80、160和320 μg/mL的姜酮单独处理没有影响IPEC-1细胞的活力(P>0.05), 160和320 μg/mL的姜酮显著缓解了3-Ac-DON引起的IPEC-1细胞活力的下降(P < 0.05)。流式细胞术检测结果显示,姜酮显著缓解了3-Ac-DON引起的IPEC-1细胞凋亡(P < 0.05),显著降低了3-Ac-DON引起的凋亡相关蛋白,如活化型半胱氨酸蛋白酶3(cleaved-Caspase-3)、B细胞淋巴瘤2相关X蛋白(BAX)和细胞色素C(CytC)的蛋白表达上调(P < 0.05)。2)蛋白免疫印迹结果显示,与3-Ac-DON处理相比,姜酮显著降低了转录活化因子6(ATF6)、磷酸化的需肌醇酶1α(p-IRE1α)和CCAAT/增强子结合蛋白同源蛋白(CHOP)的蛋白表达(P < 0.05)。3)与3-Ac-DON处理相比,姜酮显著提高了单层肠上皮细胞电阻(P < 0.05),显著降低了其通透性(P < 0.05),显著提高了闭合蛋白(occludin)和封闭蛋白-4(claudin-4)的蛋白表达(P < 0.05),但对3-Ac-DON诱导的闭锁小带蛋白-1(ZO-1)、闭锁小带蛋白-2(ZO-2)、闭锁小带蛋白-3(ZO-3)和封闭蛋白-1(claudin-1)蛋白表达下降无显著影响(P>0.05)。由此可见,姜酮缓解了3-Ac-DON引起的IPEC-1细胞凋亡、内质网应激和肠屏障功能障碍。

本文引用格式

张同坤 , 贾海 , 季昀 , 杨鹰 , 武振龙 . 姜酮缓解3-乙酰呕吐毒素诱导的猪肠上皮细胞损伤研究[J]. 动物营养学报, 2022 , 34(10) : 6695 -6701 . DOI: 10.3969/j.issn.1006-267x.2022.10.062

Abstract

The aim of this study was to investigate the effects of zingerone on the injury of porcine intestinal epithelial cells induced by 3-acetyldeoxynivalenol (3-Ac-DON). IPEC-1 cells were treated with 16 μmol/L 3-Ac-DON and 0, 80, 160 and 320 μg/mL zingerone for 24 h to select the appropriate zingerone concentration, then the effects of zingerone on apoptosis, endoplasmic reticulum stress-related proteins and tight junction proteins were studied. The results showed as follows: 1) 80, 160 and 320 μg/mL zingerone alone did not affect the viability of IPEC-1 cells (P>0.05), but 160 and 320 μg/mL zingerone significantly alleviated the decreased viability of IPEC-1 cells induced by 3-Ac-DON (P < 0.05). Flow cytometry results showed that zingerone significantly alleviated the apoptosis of IPEC-1 cells induced by 3-Ac-DON (P < 0.05), and significantly reduced the up-regulated expression of apoptosis related proteins, such as cleaved-Caspase-3, B-cell lymphoma 2-associated X protein (BAX) and cytochrome C (CytC), induced by 3-Ac-DON (P < 0.05). 2) Western blot results showed that zingerone significantly decreased the protein expressions of activating transcription factor 6 (ATF6), phosphorylated inositol-requiring transmembrane kinase/endoribonuclease 1α (p-IRE1α) and CCAAT/enhancer binding protein homologous protein (CHOP) compared with 3-Ac-DON treatment (P < 0.05). 3) Compared with 3-Ac-DON treatment, zingerone significantly increased the resistance of monolayer intestinal epithelial cells (P < 0.05), significantly decreased their permeability (P < 0.05), and significantly increased the protein expressions of occludin and claudin-4 (P < 0.05), but had no significant effects on the decreased protein expressions of zonula occluden-1 (ZO-1), zonula occluden-2 (ZO-2), zonula occluden-3 (ZO-3) and claudin-1 induced by 3-Ac-DON (P>0.05). In conclusion, zingerone can alleviate the apoptosis, endoplasmic reticulum stress and intestinal barrier dysfunction in IPEC-1 cells induced by 3-Ac-DON.

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