Roselli Marianna, Finamore Alberto, Britti Maria Serena, Konstantinov Sergey R, Smidt Hauke, de Vos Willem M, Mengheri Elena
Istituto Nazionale di Ricerca per gli Alimenti e la Nutrizione, 00178 Rome, Italy.
J Nutr. 2007 Dec;137(12):2709-16. doi: 10.1093/jn/137.12.2709.
Lactobacilli have a potential to overcome intestinal disorders; however, the exact mode of action is still largely unknown. In this study, we have used the intestinal porcine intestinal IPEC-1 epithelial cells as a model to investigate a possible protective activity of a new Lactobacillus species, the L. sobrius DSM 16698(T), against intestinal injury induced by enterotoxigenic Escherichia coli (ETEC) K88 infection and the underlying mechanisms. Treatment of infected cells with L. sobrius strongly reduced the pathogen adhesion. L. sobrius was also able to prevent the ETEC-induced membrane damage by inhibiting delocalization of zonula occludens (ZO)-1, reduction of occludin amount, rearrangement of F-actin, and dephosphorylation of occludin caused by ETEC. RT-PCR and ELISA experiments showed that L. sobrius counteracted the ETEC-induced increase of IL-8 and upregulated the IL-10 expression. The involvement of IL-8 in the deleterious effects of ETEC was proven by neutralization of IL-8 with a specific antibody. A crucial role of IL-10 was indicated by blockage of IL-10 production with neutralizing anti-IL-10 antibody that fully abrogated the L. sobrius protection. L. sobrius was also able to inhibit the internalization of ETEC, which was likely favored by the leaking barrier. The protective effects were not found with L. amylovorus DSM 20531(T) treatment, a strain derived from cattle waste but phylogenetically closely related to L. sobrius. Together, the data indicate that L. sobrius exerts protection against the harmful effects of ETEC by different mechanisms, including pathogen adhesion inhibition and maintenance of membrane barrier integrity through IL-10 regulation.
乳酸杆菌有克服肠道疾病的潜力;然而,确切的作用方式仍大多未知。在本研究中,我们使用猪肠道IPEC - 1上皮细胞作为模型,来研究一种新的乳酸杆菌菌株——嗜酒乳杆菌DSM 16698(T)——对产肠毒素大肠杆菌(ETEC)K88感染诱导的肠道损伤的可能保护活性及其潜在机制。用嗜酒乳杆菌处理感染细胞可显著减少病原体的黏附。嗜酒乳杆菌还能够通过抑制闭合蛋白(ZO)-1的移位、减少闭合蛋白的量、F -肌动蛋白的重排以及ETEC引起的闭合蛋白去磷酸化,来防止ETEC诱导的膜损伤。逆转录聚合酶链反应(RT - PCR)和酶联免疫吸附测定(ELISA)实验表明,嗜酒乳杆菌可抵消ETEC诱导的白细胞介素 - 8(IL - 8)增加,并上调白细胞介素 - 10(IL - 10)的表达。用特异性抗体中和IL - 8证明了IL - 8参与了ETEC的有害作用。用中和抗IL - 10抗体阻断IL - 10的产生,完全消除了嗜酒乳杆菌的保护作用,这表明IL - 10起关键作用。嗜酒乳杆菌还能够抑制ETEC的内化,而这可能受到渗漏屏障的促进。用源于牛粪但在系统发育上与嗜酒乳杆菌密切相关的解淀粉乳杆菌DSM 20531(T)处理未发现保护作用。总之,数据表明嗜酒乳杆菌通过不同机制对ETEC的有害作用发挥保护作用,包括抑制病原体黏附以及通过调节IL - 10维持膜屏障完整性。