Title : Beneficial effects of probiotic supplementation (Lactobacillus Plantarum Dad-13) on Body Weight, Liver Function, and Liver Histopathological Features in Non-Alcoholic Fatty Liver Disease (NAFLD) Model Sprague-Dawley Rats Disease (NAFLD) Model Sprague-Dawley Rats
Abstract:
Background:
Non-alcoholic fatty liver disease (NAFLD) is a growing global health concern, characterized by the accumulation of excess fat in the liver. The human microbiome, the vast community of microorganisms living in our bodies, plays a crucial role in various physiological processes, including liver health. Non-alcoholic fatty liver disease (NAFLD) is a prevalent liver disorder characterized by hepatic fat accumulation unrelated to alcohol consumption, with its prevalence rising alongside obesity rates. The "gut-liver" axis reveals that gut microbiota and metabolites significantly impact NAFLD development and progression.
Objective:
This study aimed to investigate the effects of probiotic Lactobacillus plantarum Dad-13 on body weight, liver function, and histopathological features in rat models of NAFLD.
Methods:
This is an experimental study with a post-test-only control group design where probiotic supplementation of L. plantarum Dad-13 was given to NAFLD model Sprague-Dawley rats fed by a high-fat-fructose diet. Twenty male rats (Rattus novergicus strain Sprague-Dawley) aged 8–12 weeks. After acclimatization, the rats were randomly divided into the standard diet group (n=4) and the high-fat-fructose diet group (n=16) for 12th weeks to induce NAFLD. On week 13th, the high-fat-fructose diet group (HFFr) was divided into two groups (n=8, each): the HFFr group fed only a high-fat-fructose diet, and the HFFr+Probiotic fed a high-fat-fructose diet with probiotic supplementation for six weeks. The experimental protocol involved administering probiotic Lactobacillus plantarum Dad-13 at a dose of 3x109 CFU/g over six weeks to the rats with NAFLD induced by a high-fat and high-fructose (HFFr) diet. Lipopolysaccharide-binding protein (LBP) serum levels were measured using the Rat LBP ELISA Kit. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured with Horiba Pentra 400 Chemistry Analyzer. The liver section was mounted on microscope slides and stained using Hematoxylin and Eosin (HE). To determine the NAFLD Activity Score (NAS), which consists of steatosis, lobular inflammation, and balloon degeneration grade, a histological examination was performed blinded by a pathologist.
Results:
Significant reductions in body and liver weight, improved liver function (serum LBP, AST, and ALT levels), and Non-alcoholic liver Activity Score (NAS) in rats fed by HFFr diets supplemented with probiotics. The serum levels of LBP in rats were received a HFFr plus probiotics diet (37.81 ± 5.45 EU/L) almost resemble the standard diet group (36.07 ± 5.60 gr EU/L), while the level of rats receiving HFFr were the highest (69.45 ± 14.90 EU/L) compared to the other groups (p<0.001 and p<0.001, respectively).
Conclusion:
This study found that supplementation with the probiotic strain effectively reduced body weight, improved liver function through decreased LBP, AST, and ALT serum levels, and improved liver histopathology features in diet-induced NAFLD. These results suggest that L. plantarum Dad-13, derived from the traditional Indonesian fermented milk product Dadih, could be a promising complementary therapeutic strategy for NAFLD management in clinical settings
