β-防御素yTAP对小鼠肠道形态及菌群多样性的影响

doi:10.16431/j.cnki.1671-7236.2025.05.040

摘要/Abstract

摘要： 【目的】探究β-防御素牦牛气管抗菌肽(yak tracheal antimicrobial peptide，yTAP)对小鼠肠道形态和微生物菌群多样性的影响，为揭示yTAP对肠道健康的作用及将其开发为新型抗菌制剂提供参考。【方法】选取24只4周龄无特定病原菌(SPF)ICR小鼠，随机分为3组：对照组(CON)、5 mg/kg yTAP组(LOW)和10 mg/kg yTAP组(HIGH)(n=8)。试验组小鼠尾静脉注射相应浓度yTAP，对照组小鼠注射等体积无菌生理盐水，0.5 mL/只。给药后正常饲养7 d，称重后处死小鼠迅速收集各脏器组织并称重，计算脏器系数。采集回肠组织，通过苏木精-伊红(HE)染色观察肠道形态变化。收集小鼠结肠内容物，提取基因组DNA并基于NovaSeq 6000测序平台进行16S rRNA扩增并测序分析。【结果】5和10 mg/kg yTAP对小鼠体重及脏器系数均无显著影响(P＞0.05)。与对照组相比，10 mg/kg yTAP组小鼠回肠隐窝深度显著降低(P＜0.05)，绒毛高度/隐窝深度比值显著升高(P＜0.05)；5 mg/kg yTAP组小鼠结肠菌群的Shannon指数显著降低(P＜0.05)，10 mg/kg yTAP组小鼠结肠菌群的Shannon指数和Simpson指数均显著降低(P＜0.05)。在门水平上，与对照组相比，10 mg/kg yTAP组小鼠结肠中厚壁菌门相对丰度及厚壁菌门/拟杆菌门比值(F/B)显著降低(P＜0.05)。在属水平上，与对照组相比，5 mg/kg yTAP组小鼠结肠的乳杆菌属相对丰度增加(P＞0.05)，10 mg/kg yTAP组小鼠结肠的臭气杆菌属相对丰度显著降低(P＜0.05)。与5 mg/kg yTAP组相比，10 mg/kg yTAP组小鼠结肠的乳杆菌属和拟杆菌属相对丰度显著降低(P＜0.05)。LEfSe分析结果显示，5 mg/kg yTAP组小鼠结肠的优势菌为乳杆菌属和拟杆菌属，10 mg/kg yTAP组小鼠结肠中的优势菌为葡萄球菌目。【结论】本试验条件下，yTAP对小鼠无急性毒性，其中10 mg/kg yTAP可改善小鼠回肠黏膜形态，显著降低结肠菌群的Alpha多样性；5 mg/kg yTAP能提高小鼠结肠乳杆菌属、拟杆菌属等优势菌属的相对丰度。

关键词: β-防御素; 牦牛气管抗菌肽(yTAP); 肠道形态; 肠道菌群

Abstract: 【Objective】 This study aimed to investigate the effects of β-defensin yak tracheal antimicrobial peptide (yTAP) on intestinal morphology and microbiota diversity in mice,in order to reveal the effect of yTAP on intestinal health and provide a reference for developing yTAP as novel antimicrobial agent.【Method】 Twenty-four 4-week-old ICR mice without specific pathogen (SPF) were randomly divided into 3 groups:Control group (CON),5 mg/kg yTAP group (LOW) and 10 mg/kg yTAP group (HIGH) (n=8).Mice in the experimental groups were injected with corresponding concentration of yTAP via tail vein,and mice in the control group were injected with equal volume sterile saline (0.5 mL/mouse).After administration,the mice were fed normally for 7 days,and then killed after weighing,organ tissues were collected and weighed quickly,and organ coefficients were calculated.The intestinal morphology was observed by hematoxylin-eosin (HE) staining.Colon contents of mice were collected,genomic DNA was extracted and 16S rRNA was amplified and sequenced based on NovaSeq 6000 sequencing platform.【Result】 There was no significant effect of 5 and 10 mg/kg yTAP on body weight and organ coefficients of mice (P＞0.05).Compared with control group,the crypt depth of ileum in 10 mg/kg yTAP group was significantly decreased (P＜0.05),and the ratio of villus height to crypt depth was significantly increased (P＜0.05).The Shannon index of colon microflora in 5 mg/kg yTAP group was significantly decreased (P＜0.05),and the Shannon and Simpson indices of colon microbiota in 10 mg/kg yTAP group were significantly decreased (P＜0.05).At the phylum level,compared with control group,the relative abundance of Firmicutes and the ratio of Firmicutes to Bacteroideta (F/B) in the colon of 10 mg/kg yTAP group were significantly decreased (P＜0.05).At the genus level,compared with control group，the relative abundance of Lactobacillus in the colon of 5 mg/kg yTAP group increased (P＞0.05).The relative abundance of Odoribacter in the colon of 10 mg/kg yTAP group was significantly decreased (P＜0.05).Compared with 5 mg/kg yTAP group,the relative abundance of Lactobacillus and Bacteroides in the colon of mice in 10 mg/kg yTAP group was significantly decreased (P＜0.05).LEfSe analysis results showed that the dominant bacteria in the colon of mice in 5 mg/kg yTAP group were Lactobacillus and Bacteroides,and the dominant bacteria in the colon of mice in 10 mg/kg yTAP group were Staphylococcales.【Conclusion】 Under this experimental conditions,yTAP had no acute toxicity to mice,among which 10 mg/kg yTAP could improve the morphology of ileum mucosa and significantly reduce the Alpha diversity of microbiota.5 mg/kg yTAP could increase the relative abundance of dominant strains such as Lactobacillus and Bacteroides in mice.

Key words: β-defensin; yak tracheal antimicrobial peptide (yTAP); intestinal morphology; intestinal microbiota

中图分类号:

S859.79

黎小银, 刘萌, 武新学, 罗璠, 高彦华. β-防御素yTAP对小鼠肠道形态及菌群多样性的影响[J]. 中国畜牧兽医, 2025, 52(5): 2379-2391.

LI Xiaoyin, LIU Meng, WU Xinxue, LUO Fan, GAO Yanhua. Effects of β-defensin yTAP on Intestinal Morphology and Microbiota Diversity in Mice[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(5): 2379-2391.

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