单宁酸对低剂量T-2毒素诱导小鼠结肠黏膜损伤与菌群失调的保护效应

doi:10.11843/j.issn.0366-6964.2023.08.041

摘要/Abstract

摘要： 旨在探究单宁酸（tannic acid，TA）对低剂量T-2毒素诱导小鼠结肠黏膜损伤与其菌群失调的保护机制。试验选取36只6周龄雄性C57BL/6J小鼠，随机分为3组，每组12只小鼠，分别为Ctrl组（空白对照）、T2组（1 mg·kg^-1 T-2毒素攻毒）和T2TA组（1 mg·kg^-1攻毒+100 mg·kg^-1 TA干预），T-2毒素灌胃每周3次，TA灌胃每天1次，试验期10周。试验期结束后，取小鼠的结肠组织和结肠内容物。苏木精-伊红染色和阿利新蓝-过碘酸雪夫氏染色观察结肠形态结构的改变；免疫组化染色观察结肠组织中Occludin、Claudin1、ZO-1蛋白含量的变化；RT-qPCR检测结肠组织中Occludin、Claudin1、Muc1、Muc2、Zo-1基因的表达量变化；Western blot检测结肠组织中Occludin、Claudin1蛋白的表达量变化；16S rRNA测序分析小鼠结肠菌群组成的差异。结果表明：1）低剂量T-2毒素不会引起小鼠结肠组织显著的形态学损伤和炎症反应。2）与Ctrl组相比，T2组小鼠结肠中杯状细胞的数量极显著减少（P<0.01），而T2TA组显著增加了小鼠杯状细胞的数量（P<0.05）。3）与Ctrl组相比，T2组极显著降低了黏蛋白Muc1、紧密连接蛋白Occludin、Claudin1和Zo-1 mRNA的表达水平（P<0.01），而T2TA组极显著增加了三者的表达水平（P<0.01）。4）与Ctrl组相比，T2组显著降低了小鼠Occludin蛋白的表达水平（P<0.05），极显著降低了Claudin1蛋白的表达水平（P<0.01），而T2TA组极显著增加了两者的表达水平。5） T-2毒素导致小鼠结肠菌群的多样性发生变化，TA的干预，可使T2组小鼠结肠中的有害菌Patescibacteria门、Saccharimonadales目、Candidatus-Saccharimonas属、unclassified_f__Ruminoccaceae属、臭气杆菌属以及嗜胆菌属丰度显著（P<0.05）或极显著（P<0.01）降低，有益微生物颤杆菌属和norank_f__Oscillospiraceae属丰度分别显著（P<0.05）和极显著（P<0.01）升高。综上所述，低剂量T-2毒素暴露会导致小鼠结肠黏膜屏障受损和结肠菌群失调，TA干预可以缓解T-2毒素引起的小鼠结肠黏膜损伤，调节结肠菌群结构，改善小鼠肠道健康。

关键词: T-2毒素, 单宁酸, 抗营养因子, 肠道屏障, 肠道菌群

Abstract: T-2 toxin is the most toxic trichothecene mycotoxin and can be present in many human foods and animal feeds. Currently, most of the studies related to the toxicology of T-2 toxin based on animal models are acute attack tests, but the contamination level of T-2 toxin in feed materials and livestock and poultry compound feeds is mostly low-level contamination. In this study, C57BL/6J mice were treated with 1 mg·kg^-1 BW T-2 toxin three times a week to assess its effects on the colonic barrier and the colonic flora. 100 mg·kg^-1 BW tannic acid (TA) was applied every day to ameliorate the toxicity of the T-2 toxin. T-2 toxin did not cause significant morphological damage and marked inflammatory responses in colonic tissue but significantly decreased the number of goblet cells in colon tissue (P<0.01). T-2 significantly downregulated the expression of mucoprotein Muc1 and the tight junction proteins Occludin, Claudin1, and ZO-1 (P<0.01). T-2 toxin changes the diversity of colonic flora in mice, the intervention of TA can make harmful bacteria in the colon of T2 group mice, such as Patescibacteria, Saccharimonadales, Candidatus-Saccharimonas, Odoribacter, Bilophila, and unclassified_f__Ruminoccaceae decreased significantly (P<0.05) or extremely significantly (P<0.01), while the abundance of beneficial microorganisms norank_f__Oscillospiraceae and Oscillibacter increased significantly (P<0.05) and extremely significantly (P<0.01) respectively. In summary, a low-dose T-2 toxin exposure will lead to the impaired colonic barrier and dysbiosis of colonic flora in mice, and TA treatment can alleviate T-2 toxin-induced colonic mucosal damage and improve intestinal health in mice.

Key words: T-2 toxin, tannic acid, antinutritional factors, intestinal barrier, intestinal flora

中图分类号:

S856.9

谢旖, 邹郦睿, 陶冉, 刘莎, 王江萍, 文利新, 邬静, 王吉. 单宁酸对低剂量T-2毒素诱导小鼠结肠黏膜损伤与菌群失调的保护效应[J]. 畜牧兽医学报, 2023, 54(8): 3582-3594.

XIE Yi, ZOU Lirui, TAO Ran, LIU Sha, WANG Jiangping, WEN Lixin, WU Jing, WANG Ji. Protective Effect of Tannic Acid on Colonic Mucosal Damage and Microflora Disturbance Induced by Low-Dose T-2 Toxin in Mice[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3582-3594.

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