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目的:验证苯扎氯铵制作的大鼠先天性巨结肠模型是否存在肠道菌群变化并探索其规律。方法:按随机数字表法将18只200 g左右SPF级Sprague-Dawley(SD)雌鼠随机划分为实验组和对照组，每组各9只，实验组采用0.1%-30 min苯扎氯铵液制作先天性巨结肠模型，对照组用无菌生理盐水处理。分别于术后2、3、4周采集上述模型鼠狭窄段、扩张段以及对照组肠段粪菌，提取粪菌DNA。16S rRNA V3-V4区扩增后，利用Illumina MiSeq测序平台检测，并进行基于OUTs的聚类分析，门水平、属水平菌群相对丰度及多样性分析。以α＝0.05为显著性水平，采用One-way ANOVA和Bonferroni校正的事后多重检验，或Kruskal-Wallis法进行差异性分析。结果:实验组狭窄段、扩张段以及对照组肠道菌群总OUTs分别为876.33±27.21、913.00±43.51和6 193.33±1.53，特有OUTs分别为52.00±16.46、64.67±3.78、5 375.33±60.12，各组上述指标与对照组比较，差异均有统计学意义(n P均<0.000 1)。实验组狭窄段、扩张段与对照组菌群ACE指数分别为1 040.15±44.2、959.30±27.95和11 923.52±36.66，Chao1指数分别为1 087.47±130.35、946.63±29.56和10 133.08±69.88，各组上述指标与对照组比较，差异均有统计学意义(n P均<0.000 1)。实验组狭窄段、扩张段以及对照组菌群Shannon值分别为6.46±0.52、6.50±0.37和8.18±0.01，各组与对照组比较，差异均有统计学意义(n P均<0.05)。对照组及实验组狭窄段、扩张段菌群门水平相对丰度较多的是厚壁菌门[(38.46±0.03)%、(72.68±6.15)%、(63.98±7.22)%]、拟杆菌门[(26.76±0.05)%、(24.91±5.98)%、(33.17±6.92)%]和变形菌门[(32.15±0.09)%、(0.70±0.30)%、(0.56±0.14)%]，除拟杆菌门外，各组与对照组比较，差异均有统计学意义(n P均<0.05)。实验组与对照组菌群属水平相对丰度前10的菌群比较，差异亦有统计学意义(n P均<0.05)。n 结论:苯扎氯铵制作的大鼠先天性巨结肠模型存在肠道菌群变化。“,”Objective:To verify whether the change of intestinal microbiome existed in rat model of benzalkonium chloride-treated Hirschsprung's disease and to explore its recurring patterns.Methods:Eighteen SPF Sprague-Dawley (SD) female rats about 200 g were randomly divided into experimental and control groups (n=9 each). Serosal application of 0.1%-30 min benzalkonium chloride was applied for establishing a model of Hirschsprung's disease in experimental group while control group received only sterile saline. Feces in stricture and expansion of both groups were collected at Week 2/3/4 postoperatively. And fecal DNA was extracted. The v3-v4 region of bacterial 16S rRNA was selected for gene amplication and sequencing using Illumina sequencing platform for OTUs, the relative abundance differences at genus/phylum level as well as diversity analysis. One-way ANOVA and Bonferroni corrected multiple tests or Kruskal-Wallis method were employed for difference analysis with α=0.05 as the significance level.Results:The fecal OTUs in stricture/expansion of experimental and control group were 876.33±27.21, 913.00±43.51 and 6 193.33±1.53. And the specific OUTs were 52.00±16.46, 64.67±3.78 and 5 375.33±60.12. The differences had statistical significance (all n P<0.000 1). Significant differences in ACE/Chao1 existed between stricture/expansion of experimental and control groups (1040.15±44.2 vs. 959.30±27.95 vs. 11 923.52±36.66, 1 087.47±130.35 vs. 946.63±29.56 vs. 10 133.08±69.88, alln P<0.0001). The Shannon values of stricture/ expansion of experimental and control groups were (6.46±0.52, 6.50±0.37 & 8.18±0.01). And there were significant differences (alln P<0.05). Firmicutes, Bacteroidetes and Proteobacteria were the most three abundant species at the phylum level in stricture/expansion of experimental and control groups. The relative abundance of the above species was different (38.46%±0.03%, 26.76%±0.05%, 32.15%±0.09% vs. 72.68%±6.15%, 24.91%±5.98%, 0.70%±0.30% vs. 63.98%±7.22%, 33.17%±6.92%, 0.56%±0.14%). Except for Bacteroidetes, the differences were statistically significant (alln P<0.05). Differences existed in the relative abundance of species at the phylum level in experimental and control groups. And the relative abundance of species at the phylum level changed with time in experimental group.n Conclusions:The change of intestinal micro-biome existed in rat model of benzalkonium chloride-treated Hirschsprung's disease.