高通量测序分析Nisin对腹泻小鼠肠道菌群的影响

doi:10.11843/j.issn.0366-6964.2018.09.023

摘要/Abstract

摘要：

采用高通量测序研究乳酸链球菌素（Nisin）对腹泻小鼠肠道菌群结构的影响。通过腹腔灌注E.coli O₁建立腹泻病小鼠模型，通过动物体内保护率试验筛选出最佳药物剂量，将小鼠随机分为空白对照组、阴性对照组、氨苄青霉素、盐酸环丙沙星组和Nisin组。连续灌胃15 d，收集盲肠粪便，采用高通量测序对5组样品进行分析。结果表明：1）E.coli O₁可使腹泻小鼠十二指肠绒毛断裂；Nisin可增加淋巴细胞百分比，起到增强免疫的目的。2）通过小鼠保护率试验筛选出Nisin最佳剂量（0.002 g·mL^-1），其保护率为37.5%，低于氨苄青霉素（50%）和盐酸环丙沙星组（50%）。3）通过盲肠菌落计数，Nisin中大肠杆菌和肠球菌的数量显著低于阴性对照组（P<0.05），尤其以Nisin组中肠球菌的数量最低；Nisin组中乳酸杆菌的数量显著高于其他各组（P<0.05）。4）Nisin组中样品菌群多样性指数（ACE=1 417.25，Chao1=1 378.45，Shannon=7.56）显著高于阴性对照组（ACE=969.54，Chao1=340.29，Shannon=6.63），另外，腹泻小鼠肠道菌群结构存在显著差异。5）在门和属水平上，Nisin组中Bacteroidetes（拟杆菌门）最低，Firmicutes（厚壁杆菌门）、Clostridium（梭状芽胞杆菌）、Akkermansia属和Lactobacillus（乳杆菌属）所占比例最高，且高于其他各组。Nisin可增强腹泻小鼠免疫功能，增加肠道内有益菌的数量，减少有害菌数量；摄入适量的Nisin可以调控肠道内菌群比例。

Abstract:

In this study, high-throughput sequencing was used to study the effect of Nisin on intestinal microflora in diarrhea mice. The model of diarrhea mice was established by peritoneal perfusion of E. coli O₁, and the best dose of the drug was screened by animal protection test in vivo. The mice were randomly divided into blank control group, negative control group, ampicillin, ciprofloxacin hydrochloride group and Nisin group. Gavage was continued for 15 days, and cecum stool were collected. Five samples were analyzed by high-throughput sequencing. The results showed that:1) E. coli O₁ can duodenal villi in diarrhea mice. Nisin increased the percentage of lymphocytes, to enhance immunity. 2) The best Nisin dose (0.002 g·mL^-1) was screened by mouse protection rate test, the protection rate was 37.5%, lower than that of ampicillin (50%), ciprofloxacin hydrochloride group. 3) Counting of cecal colonies:the number of E. coli and Enterococci in Nisin was lower than that in the negative control group (P<0.05), especially in Nisin group. The number of Lactobacillus in Nisin group was higher than that in other group (P<0.05). 4)The diversity index (ACE=1 417.25, Chao1=1 378.45, Shannon=7.56) in Nisin group was higher than that of negative control group (ACE=969.54, Chao1=340.29, Shannon=6.63). There are significant differences in the flora structure. 5) The Bacteroidetes were the lowest in the Nisin group, with the highest proportion of Firmicutes, Clostridium, Akkermansia and Lactobacillus being higher than Other groups. Therefore, Nisin enhanced the immune function of diarrhea mice, increased the number of beneficial bacteria in the intestine and reduced the number of harmful bacteria; intake of appropriate amount of Nisin can regulate the proportion of intestinal flora.

中图分类号:

S855.12

贺美玲, 王纯洁, 贾知锋, 斯木吉德, 敖日格乐. 高通量测序分析Nisin对腹泻小鼠肠道菌群的影响[J]. 畜牧兽医学报, 2018, 49(9): 2015-2024.

HE Mei-ling, WANG Chun-jie, JIA Zhi-feng, SI-mu-ji-de, AO-ri-ge-le. The Effect of Nisin on Intestinal Flora in Diarrheal Mice Analyzed by High-throughput Sequencing[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(9): 2015-2024.

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