Department of Aquatic Animal Medicine, College of Fisheries, Huazhong Agricultural University, Wuhan, 430070, China.
Key Lab of Freshwater Animal Breeding, Ministry of Agriculture, Wuhan, 430070, China.
Sci Rep. 2017 May 8;7(1):1588. doi: 10.1038/s41598-017-01336-9.
Our study explored the effect of oral intubation of Bacillus subtilis on Aeromonas hydrophila-induced intestinal mucosal barrier function damage and inflammation in grass carp. The mid-intestine mucosal tissue was collected for ATPase activity measurement. Intestinal mucosa was also ultrastructurally examined with transmission electron microscope (TEM), and its permeability was determined using Evans blue (EB) and D-lactic acid. The mid-intestine pro-inflammation cytokine, MyD88 and tight junction (TJ) protein mRNA expression levels were measured using real-time quantitative PCR. The results revealed that B. subtilis was found to prevent the decrease in the activity of Na, K-ATPase and Ca, Mg-ATPase, as well as the increase in EB and D-lactic acid concentration and inflammation induced by A. hydrophila in grass carp. Compared with A. hydrophila groups, B. subtilis safeguarded the integrity of intestinal villi and tight junction structure and restrained A. hydrophila-induced down-regulation of TJ proteins zonula occludens-1 (ZO-1) and occludin. B. subtilis also restrained up-regulation of TJ protein claudin b, pro-inflammation cytokine tumour necrosis factor α (TNF-α), cytokine interleukin 8 (IL-8), IL-1β, and adaptor protein myeloid differentiation factor 88 (MyD88) mRNA levels. Thus, oral intubation of B. subtilis could reduce A. hydrophila-induced intestinal mucosal barrier function damage and inflammation.
我们的研究探讨了枯草芽孢杆菌经口灌胃对嗜水气单胞菌诱导的草鱼肠道黏膜屏障功能损伤和炎症的影响。采集中肠黏膜组织用于测量 ATP 酶活性。用透射电子显微镜(TEM)超微结构观察肠黏膜,用伊文思蓝(EB)和 D-乳酸测定其通透性。用实时定量 PCR 测定中肠促炎细胞因子 MyD88 和紧密连接(TJ)蛋白 mRNA 表达水平。结果表明,枯草芽孢杆菌可防止 Na,K-ATP 酶和 Ca,Mg-ATP 酶活性下降,以及 EB 和 D-乳酸浓度升高和嗜水气单胞菌诱导的草鱼炎症。与嗜水气单胞菌组相比,枯草芽孢杆菌保护了肠绒毛的完整性和紧密连接结构,并抑制了 TJ 蛋白闭锁蛋白-1(ZO-1)和闭合蛋白的下调。枯草芽孢杆菌还抑制了 TJ 蛋白 Claudin b、促炎细胞因子肿瘤坏死因子-α(TNF-α)、细胞因子白细胞介素 8(IL-8)、IL-1β 和衔接蛋白髓样分化因子 88(MyD88)mRNA 水平的上调。因此,枯草芽孢杆菌经口灌胃可以减轻嗜水气单胞菌诱导的草鱼肠道黏膜屏障功能损伤和炎症。