摘要
目的 探讨脱氧胆酸钠诱导型慢性胃炎Wistar大鼠肠道菌群结构的变化。方法 将40只SPF级Wistar雄性大鼠随机分为对照组(CG组)和模型组(MG组),每组20只;MG组以20 mmol/L脱氧胆酸钠[2 mL/(只·d)]、60%的乙醇[1 mL/(只·3 d)]灌胃以及0.03%和0.05%氨水日常饮用,同时联合饥饱失常(2 d饱食1 d禁食),持续处理12周后,解剖大鼠,裸眼和H&E染色观察胃组织病理学变化;采用高通量测序和生物信息学技术分析肠道菌群结构和多样性改变。结果 与CG组大鼠比较,裸眼观察,MG组胃腔扩张,壁变薄,皱襞少而浅,胃黏膜略显苍白、色浅;H&E染色表明,胃黏膜腺体扩张,排列稀疏紊乱,上皮细胞排列不规则,伴坏死脱落,胃小凹结构破坏。Alpha多样性分析显示,Chao1、Ace和Shannon指数显著降低,Simpson指数显著升高,差异有统计学意义(t=11.217 3、10.615 2、7.981 8、12.684 9,均P<0.001)。从门水平而言,MG组大鼠肠道菌群中Actinobacteria(t=2.318 7,P=0.031 7)、Cyanobacteria(t=1.169 6,P=0.256 6)、Proteobacteria(t=2.178 6,P=0.042 2)、Firmicutes(t=0.329 8,P=0.745 1)和Verrucomicrobia(t=1.190 1,P=0.248 6)细菌数量增加;Bacteroidetes(t=1.918 4,P=0.070 2)、Patescibacteria(t=0.135 8,P=0.893 4)和Tenericutes(t=1.419 9,P=0.171 8)细菌数量下降。从属水平而言,MG组大鼠肠道菌群Firmicutes中的Eubacterium oxidoreducens group, Proteobacteria中的Aliihoeflea、Proteus,Actinobacteria中的Eggerthellaceae;nclassified的相对丰度较CG组增高(t=2.232 9,P=0.037 8;t=3.020 8,P=0.007 0;t=2.286 1,P=0.033 9;t=2.330 7,P=0.030 9);Firmicutes中的Eubacterium xylanophilum group、Acetitomaculum、Lachnospiraceae NC2004 group、Globicatella、Anaerovorax、Lachnospiraceae A2和Lachnoclostridium 5,Proteobacteria中的Citrobacter的相对丰度低于CG组(t=3.101 0,P=0.005 8;t=2.620 3,P=0.016 8;t=2.509 6,P=0.021 3;t=2.498 3,P=0.021 8;t=2.417 5,P=0.025 8;t=2.339 5,P=0.030 3;t=2.315 1,P=0.031 9;t=2.158 2,P=0.043 9)。结论 脱氧胆酸钠、乙醇、氨水和饥饱失常联合的慢性胃炎建模法可引起Wistar大鼠肠道菌群结构发生改变。
Objective To explore the changes of intestinal flora structure in Wistar rats with chronic gastritis induced by deoxycholic acid(DA). Methods Forty SPF Wistar rats were randomly divided into control group(CG) and model group(MG), with 20 rats in each group. The model group were treated with 20 mmol/L sodium deoxycholate [2 mL/(rat·d)] and 60% ethanol [1 mL/(rat·3 d)] by gavage, 0.03% and 0.05% ammonia water by daily drinking, and the combination of starvation and satiety disorder(2 days of satiety and 1 day of starvation) to establish animal models of chronic gastritis. After 12 weeks, the rats were dissected. The gastric histopathological changes were observed with naked eyes and H&E staining. The changes in the structure and diversity of intestinal flora in the faeces were analyzed with high-throughput sequencing and bioinformatics technology. Results Compared with CG, characteristic changes were observed in MG by naked eyes, including dilated gastric cavity, thinner wall, fewer and lighter folds, slightly pale gastric mucosa with light color. H&E staining showed that the gastric mucosa glands were dilated, sparse, disordered and irregular, epithelial cells were arranged irregularly, accompanied with necrosis and shedding, and the structure of gastric fovea was destroyed. Alpha diversity analysis showed that indexes of Chao1, Ace and Shannon were decreased significantly, but Simpson index increased(t=11.217 3, 10.615 2, 7.981 8, 12.684 9, all P<0.001). At the phylum level, the relative abundances of Actinobacteria(t=2.318 7, P=0.031 7), Cyanobacteria(t=1.169 6,P=0.256 6), Proteobacteria(t=2.178 6, P=0.042 2), Firmicutes(t=0.329 8, P=0.745 1) and Verrucomicrobia(t=1.190 1,P=0.248 6) increased, while those of Bacteroidetes(t=1.918 4,P=0.070 2), Patescibacteria(t=0.135 8,P=0.893 4) and Tenericutes(t=1.419 9,P=0.171 8) decreased. At the genus level, the relative abundances of Eubacterium oxidoreducens group of Firmicutes, Aliihoeflea and Proteus of Proteobacteria, and Eggerthellaceae;nclassified of Actinobacteria were significantly higher in MG than those in CG(t=2.232 9, P=0.037 8;t=3.020 8, P=0.007 0;t=2.286 1, P=0.033 9;t=2.330 7, P=0.030 9), while those of Eucharacterium xylanophilum group, Acetitomaculum, Lachnospiraceae NC2004 group, Globicella, Anaerovorax, Lachnospiraceae A2, and Lachnosclostridium 5 in Firmicutes and Citrobacter in Proteobacteria were significantly lower than those in CG, respectively(t=3.101 0, P=0.005 8;t=2.620 3, P=0.016 8;t=2.509 6, P=0.021 3;t=2.498 3, P=0.021 8;t=2.417 5, P=0.025 8;t=2.339 5, P=0.030 3;t=2.315 1, P=0.031 9;t=2.158 2, P=0.043 9). Conclusion The chronic gastritis modeling with sodium deoxycholate, ethanol, ammonia and disorder of starvation and satiety can change the intestinal flora structure of Wistar rats.
作者
伍志伟
张录梅
薛娜
孙文平
周熙祥
房明
WU Zhiwei;ZHANG Lumei;XUE Na;SUN Wenping;ZHOU Xixiang;FANG Ming(Preclinical Medicine College,Gansu University of Chinese Medicine,Lanzhou,Gansu 730030,China;不详)
出处
《中国微生态学杂志》
CAS
CSCD
2022年第2期125-133,共9页
Chinese Journal of Microecology
基金
甘肃省自然科学基金(18JR3RA200)
甘肃省高校重大疾病分子医学与中医药防治研究重点实验室开放基金(FZYX15-16-1)。
