Prophylactic Treatment of Probiotic and Metformin Mitigates Ethanol-Induced Intestinal Barrier Injury: In Vitro, In Vivo, and In Silico Approaches.

Farhin Patel,Palash Mandal,Kirti Parwani,Priyashi Rao,Rakesh Rawal,Dhara Patel,Sanjoy Roychowdhury

doi:10.1155/2021/5245197

Abstract

Ethanol depletes intestinal integrity and promotes gut dysbiosis. Studies have suggested the individual role of probiotics and metformin Met in protecting intestinal barrier function from injuries induced by ethanol. The objective of the current study is to investigate the potential mechanism by which coadministration of probiotic Visbiome® (V) and Met blocks the ethanol-induced intestinal barrier dysfunction/gut leakiness utilizing Caco-2 monolayers, a rat model with chronic ethanol injury, and in silico docking interaction models. In Caco-2 monolayers, exposure to ethanol significantly disrupted tight junction (TJ) localization, elevated monolayer permeability, and oxidative stress compared with controls. However, cotreatment with probiotic V and Met largely ameliorated the ethanol-induced mucosal barrier dysfunction, TJ disruption, and gut oxidative stress compared with ethanol-exposed monolayers and individual treatment of either agent. Rats fed with ethanol-containing Lieber-DeCarli liquid diet showed decreased expression of TJ proteins, and increased intestinal barrier injury resulting in pro-inflammatory response and oxidative stress in the colon. We found that co-administration of probiotic V and Met improved the expression of intestinal TJ proteins (ZO-1 and occludin) and upregulated the anti-inflammatory response, leading to reduced ER stress. Moreover, co-administration of probiotic V and Met inhibited the CYP2E1 and NOX gene expression, and increase the translocation of Nrf-2 as well as anti-oxidative genes (SOD, catalase, Gpx, and HO-1), leading to reduced colonic ROS content and malondialdehyde levels. The combined treatment of probiotic V and Met also improved their binding affinities towards HO-1, Nrf-2, SLC5A8, and GPR109A, which could be attributed to their synergistic effect. Our findings based on in-vitro, in-vivo, and in-silico analyses suggest that the combination of probiotic V and Met potentially acts in synergism, attributable to their property of inhibition of inflammation and oxidative stress against ethanol-induced intestinal barrier injury.

Highlights

Chronic alcohol consumption is associated with numerous gastrointestinal and liver disorders, especially alcoholic liver disease (ALD) [1]
To know and understand the plausible mechanism by which combined treatment of probiotic V and Met could act in synergism to prevent oxidative stress and to maintain intestinal permeability induced by ethanol, we adopted an in silico approach and performed the docking of Met and butyrate with antioxidants, i.e., Nrf-2 and HO-1 as well as butyrate sensors, i.e., the butyrate receptor GPR109A and the butyrate transporter SLC5A8
In accordance with the above results, our results demonstrated that coadministration of probiotic V and Met significantly prevented the decreased levels of Transepithelial Electrical Resistance (TEER) and intestinal hyperpermeability induced by ethanol indicating the combinatorial treatment completely blocks the ethanol-induced intestinal barrier dysfunction

Summary

Introduction

Chronic alcohol consumption is associated with numerous gastrointestinal and liver disorders, especially alcoholic liver disease (ALD) [1]. Alcohol consumption is known to disrupt TJs resulting in increased intestinal permeability [5]. It is a most likely accepted fact that impaired intestinal epithelial integrity and intestinal. In alcoholics, increased blood endotoxin levels (endotoxemia) can be due to three potential mechanisms: (i) due to both abnormal gut dysbiosis and overproduction of bacteria leading to increased endotoxin production; (ii) due to gut leakiness, which causes increased endotoxin permeation through the gut; and (iii) due to either portal hypertension or Kupffer cell dysfunction leading to decreased endotoxin elimination [8, 9]. Gut leakiness or disrupted intestinal barrier function is considered to be the important mechanism for ethanol-induced increased endotoxemia in alcohol-mediated liver injury [10]

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