熊去氧胆酸联合双歧三联活菌对结肠炎小鼠炎症因子及肠道菌群影响

Effects of ursodeoxycholic acid combined with Bifidobacterium on inflammatory factors and intestinal flora in mice with colitis

目的探究熊去氧胆酸(UDCA)联合双歧三联活菌对溃疡性结肠炎(UC)小鼠IL-10、TLR4及肠道菌群的影响。方法购自中国科学院微生物研究所SPF级BALB/c小鼠60只,按照区间分组法将小鼠分为对照组、UC组、UDCA组、双歧三联活菌组、UDCA联合组各12只。每日观察小鼠一般情况、收集粪便做大便隐血实验检测、采用疾病活动指数(DAI)评分判断疾病活动指数;于建模成功7 d后采集各组眼眶静脉血、断头法处死后获取结肠组织,检测IL-10、TLR4,进行UC组织病理学评分,并取小鼠粪便分析其肠道菌群变化。结果第3、7、21 d UDCA组、双歧三联活菌组、UDCA联合组DAI评分均低于UC组,且UDCA联合组DAI评分低于UDCA组、双歧三联活菌组;第21 d UC组、UDCA组、双歧三联活菌组的体质量均低于对照组,UDCA组、双歧三联活菌组、UDCA联合组体质量均高于UC组,且UDCA联合组体质量高于UDCA组、双歧三联活菌组(P<0.05)。对照组组织病理学评分低于UC组、UDCA组、双歧三联活菌组、UDCA联合组,UDCA组、双歧三联活菌组、UDCA联合组均低于UC组,UDCA联合组低于UC组、UDCA组、双歧三联活菌组(P<0.05)。UC组、UDCA组、双歧三联活菌组、UDCA联合组条带数较对照组减少,UDCA组、双歧三联活菌组、UDCA联合组条带数较UC组增多,UDCA联合组条带数多于UDCA组、双歧三联活菌组(P<0.05)。UC组、UDCA组、双歧三联活菌组、UDCA联合组的肠杆菌、肠球菌、拟杆菌、梭杆菌数量均较对照组少;而UDCA组、双歧三联活菌组、UDCA联合组较UC组多,且UDCA联合组较UDCA组、双歧三联活菌组多(P<0.05)。结论熊去氧胆酸联合双歧三联活菌治疗UC小鼠,可明显降低IL-10、TLR4水平,增加肠道菌群数量,治疗效果更佳。

Objective To explore the effect of ursodeoxycholic acid(UDCA) combined with bifidobacteria on ulcerative colitis(UC) in mice based on the changes of IL-10, TLR4 and intestinal flora. Methods Female BALB/c mice(SPF grade) were divided into control group, UC group, UDCA group, Bifidobacterium group or UDCA combination group based on the interval grouping method(n=12). The general condition of mice was observed daily;feces were collected for fecal occult blood test, and disease activity index(DAI) was used to assess disease activity index. Seven days after modeling, the blood of orbital vein and colon tissue were obtained after amputation. IL-10 and TLR4 were detected. UC histopathological score was evaluated, and the intestinal flora of mice was analyzed through feces analysis. Results On Day 3, 7 and 21, the DAI scores in UDCA group, Bifidobacterium group and UDCA combination group were lower than that in UC group, and that in UDCA combination group was lower than those in UDCA group and Bifidobacterium group. On Day 21, the body weights in UC group, UDCA group and bifidobacterium group were lower than in control group, and those in UDCA group, bifidobacterium group and UDCA combination group were higher than in UC group;the body weight in UDCA combination group was higher than in UDCA group and bifidobacterium group(all P<0.05). The histopathological score in control group was lower than in UC group, UDCA group, bifidobacterium group and UDCA combination group, those in UDCA group, bifidobacterium group and UDCA combination group were lower than in UC group, while that in UDCA combination group was lower than in UC group, UDCA group and bifidobacterium group(all P<0.05). Compared with the control group, the numbers of bands in UC group, UDCA group, bifidobacterium group and UDCA combination group decreased, while those in UDCA group, bifidobacterium group and UDCA combination group increased, and that in UDCA combination group was more than in UDCA group and bifidobacterium group(all P<0.05). The counts of Enterobacteriaceae, Enterococcus, Bacteroides, and Fusobacterium in UC group, UDCA group, bifidobacterium group and UDCA combination group were less than in the control group, those in UDCA group, bifidobacterium group and UDCA combination group were more than in UC group, and that in UDCA combined group was more than in UDCA group and bifidobacterium group(Ps<0.05). Conclusion Ursodeoxycholic acid combined with bifidobacteria can significantly reduce the levels of IL-10 and TLR4 and increase the number of intestinal flora in UC mice.