Research Article Metformin and Probiotics Interplay in Amelioration of Ethanol-Induced Oxidative Stress and Inflammatory Response in an In Vitro and In Vivo Model of Hepatic Injury Farhin Patel , Kirti Parwani , Dhara Patel , and Palash Mandal Department of Biological Sciences, P. D. Patel Institute of Applied Sciences, Charotar University of Science and Technology, Changa, 388421 Anand, Gujarat, India Correspondence should be addressed to Palash Mandal; palashmandal.bio@charusat.ac.in Received 11 December 2020; Revised 26 February 2021; Accepted 14 March 2021; Published 16 April 2021 Academic Editor: Muthuraju Sangu Copyright © 2021 Farhin Patel et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Alcohol-induced liver injury implicates inflammation and oxidative stress as important mediators. Despite rigorous research, there is still no Food and Drug Administration (FDA) approved therapies for any stage of alcoholic liver disease (ALD). Interestingly, metformin (Met) and several probiotic strains possess the potential of inhibiting alcoholic liver injury. Therefore, we investigated the effectiveness of combination therapy using a mixture of eight strains of lactic acid-producing bacteria, commercialized as Visbiome® (V) and Met in preventing the ethanol-induced hepatic injury using in vitro and in vivo models. Human HepG2 cells and male Wistar rats were exposed to ethanol and simultaneously treated with probiotic V or Met alone as well as in combination. Endoplasmic reticulum (ER) stress markers, inflammatory markers, lipid metabolism, reactive oxygen species (ROS) production, and oxidative stress were evaluated, using qRT-PCR, Oil red O staining, fluorimetry, and HPLC. In vitro, probiotic V and Met in combination prevented ethanol-induced cellular injury, ER stress, oxidative stress, and regulated lipid metabolism as well as inflammatory response in HepG2 cells. Probiotic V and Met also promoted macrophage polarization towards the M2 phenotype in ethanol-exposed RAW 264.7 macrophage cells. In vivo, combined administration of probiotic V and Met ameliorated the histopathological changes, inflammatory response, hepatic markers (liver enzymes), and lipid metabolism induced by ethanol. It also improved the antioxidant markers (HO-1 and Nrf-2), as seen by their protein levels in both HepG2 cells as well as liver tissue using ELISA. Hence, probiotic V may act, in addition to the Met, as an effective preventive treatment against ethanol-induced hepatic injury. 1. Introduction Alcohol-induced hepatic injury is the main root of morbidity and mortality universally among individuals who misuse alcohol [1]. Chronic alcohol consumption eventually leads to ALD, which encompasses steatosis, alcoholic hepatitis, fibrosis, cirrhosis, and lately, hepatocellular carcinoma (HCC) [2]. Several studies reported that oxidative stress and inflammatory responses play a key role in the develop- ment and progression of alcohol-induced liver damage [3, 4]. A few of the reasons behind the progression are (a) expression of pro-inflammatory enzymes, cytokines, and chemokines through Kupffer cell activation [5] and (b) over- production of reactive oxygen species (ROS) leading to ROS- mediated liver injury [6], and this can eventually lead to oxi- dative stress (excessive ROS generation leads to increase oxi- dant formation and reduced levels of antioxidants) [7] and also lipid peroxidation (increased ROS production leads to the generation of 4-HNE which forms adducts with protein and DNA) [8]. Thus, inhibiting the inflammation and oxida- tive stress would play a major role in preventing ethanol- induced hepatic injury. Till the time, limited acceptable developments have been completed in controlling the progression of liver injury. Thus, unique and trustworthy therapeutic approaches are strongly needed. Metformin (Met) is a commonly approved antihypergly- cemic drug of the biguanide family commonly used to control type 2 diabetes. In humans, there is a resilient connection Hindawi Mediators of Inflammation Volume 2021, Article ID 6636152, 31 pages https://doi.org/10.1155/2021/6636152