You are on page 1of 5

Enhancing Probiotic Escherichia coli Nissle 1917 inhibits leaky gut by Mucosal Integrity

ABSTRACT A. Background Probiotics are proposed to positively modulate the intestinal epithelial barrier formed by intestinal epithelial cells (IECs) and intercellular junctions. Disruption of this border alters cellular permeability and is one of the reasons for the development of enteric infections and inflammatory bowel diseases (IBDs).

B. Methodology and Findings

To study the in vivo effect of E.coli Nissle 1917 (EcN0 on the stability of the internal barrier under healthy conditions, germfree mice were colonized with EcN. IECs were isolated & analyzed for gene & protein expression of the tight junction molecules ZO-1 & ZO-2. Then the probiotic was orally administered to the gnotobiotic mice with acute dextran sodium sulfate (DSS) induced colitis. Concomittant administration of EcN to DSS treated animals resulted in a significant protection against intestinal barrier dysfunction and IECs isolated from these mice showed a more pronounced expression of ZO-1.

The protein zona occludens (ZO-1) with mass ~210-225 kDa is a structural link between cytoskeleton and tight junctions. Reorganization of ZO-1 results in increased permeability during IBDs. . INTRODUCTION The epithelial layer of gastro-intestinal tract serves as one of the primary interfaces with the outside world. ZO-1 also binds ZO-2 forming a complex. Reduction in the integrity of this barrier can promote the manifestation of enteric infections and is an important feature of IBDs like Crohn’s disease and ulcerative colitis. It is in constant contact with microbes. The barrier is formed by epithelial cells and the junctional complex excludes majority of these microbes from reaching the sub-epithelial cells.C. EcN has evolved to one of the best characterized probiotic and its efficiency and safety has been proven. Tight junctions are macromolecular protein structures which seal spaces between IECs. The role of leaky gut in pathogenesis of gastro intestinal diseases is recognized. Conclusion The study has demonstrated that EcN is able to mediate up-regulation of ZO-1 expression in murine IECs & protects from DSS colitis associated increase in intestinal mucosal permeability.

RNA isolation: To analyze ZO-1 and ZO-2 mRNA expression. . Group III received drinking water with 4-6% DSS and was given 1. The supernatant was collected. rinsed with PBS & opened. total RNA was reverse transcribed. 5. Bacteria were collected by centrifugation. 4. Induction of colitis: Acute colitis was induced to mice by addition of 4-6% dextran sodium sulfate to drinking water for 8 days. 2. Group I was treated orally with PBS two times a day.MATERIAL AND METHODS 1. Mucus was removed by treatment with DTT for 15 mins at 37 degree C on a shaker. ii. The pellet was redissolved in 200ul sterile PBS and administered by oral gauge. Isolation of IECs: Small/large intestine was isolated. Bacterial colonization: EcN was grown to an OD600 from an overnight culture diluted 1:500 in LB media.5 mM EDTA & tumbled for 10 mins at 37 degree C. Mice: All mice used in this study were 6-8 weeks old females.5-2 x 10 CFU. iii. The mice were divided to 3 groupsi. It was placed in HBBS/1. 3. Group II was treated with 4-6% DSS & treated orally with PBS twice a day.

3. Elevated ZO-1 expression after EcN treatment in experimental colitis. ZO-1 mRNA protein and protein expression are elevated in mice colonized with EcN. IECs of mice with DSS-induced colitis also exhibit a pronounced expression of ZO-1 mRNA.RESULT 1. 2. EcN is capable of producing a specific up-regulation of ZO-1 expression in IECs of healthy mice. EcN provides protection against DSS mediated leakiness of gut in our mice. . DISCUSSION The current study establishes that EcN 1917 positively impacts the intestinal epithelial barrier in vivo in three ways1. 3. 2. Reduction of colonic epithelial permeability after EcN treatment in epithelial colitis. When treated concomitantly with EcN. Our data suggests that one of the protective effects of EcN treatment on colitis prevention could be due to tight junctional integrity which in turn leads to a preserved intestinal barrier function against infectious agents.