Barbaro M R, Fuschi D, Cremon C, Carapelle M, Dino P, Marcellini M M, Dothel G, De Ponti F, Stanghellini V, Barbara G
Department of Medical and Surgical Sciences (DIMEC), Center for Applied Biomedical Research (CRBA), Alma Mater Studiorum - University of Bologna and S. Orsola-Malpighi Hospital, Bologna, Italy.
Neurogastroenterol Motil. 2018 Jun 28:e13388. doi: 10.1111/nmo.13388.
Intestinal permeability is altered in a subgroup of irritable bowel syndrome (IBS) patients and may contribute to symptom development. The aim of this study was to evaluate the in vitro effect of the probiotic Escherichia coli Nissle 1917 (EcN) on Caco-2 permeability alterations induced by mediators released by IBS mucosal biopsies compared to asymptomatic controls (AC).
Caco-2 cells were used as an in vitro model of intestinal permeability. Seven AC and 28 well-phenotyped IBS (9 IBS-D, 8 IBS-C, and 11 IBS-M) patients were enrolled. Mucosal mediators spontaneously released (SUP) by IBS and AC biopsies were collected. Two concentrations of EcN (10 and 10 ) were applied to Caco-2 with or without SUP or SLIGRL (a protease-activated receptor-2 activating peptide), tumor necrosis factor-α, and interferon-γ. Paracellular permeability was assessed by evaluating the flow of sulfonic-acid conjugated to fluorescein through Caco-2 monolayer.
EcN 10 significantly reinforced Caco-2 monolayer compared to cells incubated with medium alone. IBS SUP induced a significant increase in paracellular permeability compared to AC SUP, independently of IBS bowel habit. EcN 10 induced a significant recovery of permeability rate compared to IBS SUP. Permeability increase induced by IBS SUP significantly correlated with severity and frequency of abdominal pain and abdominal distension. The co-incubation of EcN and IBS SUP abolished the above significant correlations.
EcN reinforces the integrity of Caco-2 monolayer and reverts the increase of permeability induced by mediators released by IBS biopsies. Future studies should investigate EcN therapeutic potentials in IBS.
肠易激综合征（IBS）患者亚组的肠道通透性发生改变，这可能促使症状发展。本研究的目的是评估益生菌大肠杆菌Nissle 1917（EcN）在体外对IBS黏膜活检释放的介质诱导的Caco-2通透性改变的影响，并与无症状对照（AC）进行比较。
将Caco-2细胞用作肠道通透性的体外模型。纳入7名AC和28名表型明确的IBS患者（9名腹泻型IBS、8名便秘型IBS和11名混合型IBS）。收集IBS和AC活检组织自发释放的（SUP）黏膜介质。将两种浓度的EcN（10和10 ）应用于有或无SUP或SLIGRL（一种蛋白酶激活受体-2激活肽）、肿瘤坏死因子-α和干扰素-γ的Caco-2细胞。通过评估与荧光素偶联的磺酸通过Caco-2单层的流量来评估细胞旁通透性。
与仅用培养基孵育的细胞相比，EcN 10显著增强了Caco-2单层。与AC SUP相比，IBS SUP导致细胞旁通透性显著增加，与IBS的排便习惯无关。与IBS SUP相比，EcN 10导致通透性速率显著恢复。IBS SUP诱导的通透性增加与腹痛和腹胀的严重程度及频率显著相关。EcN与IBS SUP共同孵育消除了上述显著相关性。
EcN增强了Caco-2单层的完整性，并逆转了IBS活检组织释放的介质诱导的通透性增加。未来的研究应调查EcN在IBS中的治疗潜力。
