In Vitro and In Vivo Protective Effects of Lentil (Lens culinaris) Extract against Oxidative Stress-Induced Hepatotoxicity

Yeon-Seop Jung,Syng-Ook Lee,So Young Chun,Byung Ik Jang,Dae Hwan Kim,Man-Hoon Han,So-Hee Lee

doi:10.3390/molecules27010059

Abstract

Excessive oxidative stress plays a role in hepatotoxicity and the pathogenesis of hepatic diseases. In our previous study, the phenolic extract of beluga lentil (BLE) showed the most potent in vitro antioxidant activity among extracts of four common varieties of lentils; thus, we hypothesized that BLE might protect liver cells against oxidative stress-induced cytotoxicity. BLE was evaluated for its protective effects against oxidative stress-induced hepatotoxicity in AML12 mouse hepatocytes and BALB/c mice. H2O2 treatment caused a marked decrease in cell viability; however, pretreatment with BLE (25–100 μg/mL) for 24 h significantly preserved the viability of H2O2-treated cells up to about 50% at 100 μg/mL. As expected, BLE dramatically reduced intracellular reactive oxygen species (ROS) levels in a dose-dependent manner in H2O2-treated cells. Further mechanistic studies demonstrated that BLE reduced cellular ROS levels, partly by increasing expression of antioxidant genes. Furthermore, pretreatment with BLE (400 mg/kg) for 2 weeks significantly reduced serum levels of alanine transaminase and triglyceride by about 49% and 40%, respectively, and increased the expression and activity of glutathione peroxidase in CCl4-treated BALB/c mice. These results suggest that BLE protects liver cells against oxidative stress, partly by inducing cellular antioxidant system; thus, it represents a potential source of nutraceuticals with hepatoprotective effects.

Highlights

IntroductionLiver diseases have become a major global public health problem, and severe liver injury can overwhelm the self-regenerative capacity of the liver and can prove fatal [1]
Liver diseases have become a major global public health problem, and severe liver injury can overwhelm the self-regenerative capacity of the liver and can prove fatal [1].Excessive oxidative stress plays a role in the pathogenesis of hepatic diseases such as alcoholrelated liver disease, non-alcoholic fatty liver disease, hepatitis, fibrosis, and cirrhosis and can be induced by intrinsic factors and/or multiple xenobiotics such as alcohol and toxic compounds [1,2,3,4].Living organisms produce reactive oxygen species (ROS) and reactive nitrogen species (RNS) via normal cellular metabolism [5]
We further investigated whether beluga lentil (BLE) can protect AML12 cells from cytotoxicity induced by free fatty acids (FFA; palmitic acid:oleic acid/1:1), which is a different type of pro-oxidant known to induce hepatotoxicity [7,11]

Summary

Introduction

Liver diseases have become a major global public health problem, and severe liver injury can overwhelm the self-regenerative capacity of the liver and can prove fatal [1]. ROS and RNS generated by various pro-oxidants including alcohol [6], acetaminophen [2,4], ethyl carbamate [3], and fatty acids [7] are scavenged by various cellular enzymatic and nonenzymatic antioxidants, such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione (GSH), and thioredoxin. These cellular antioxidants possibly play an important role in the prevention of oxidative stressrelated diseases, such as liver disease [8]. In vivo hepatoprotective effects of BLE were further evaluated in an animal model of CCl4 -induced acute liver failure

Materials and Preparation of Lentil Extract

Cell Cultures

Animals and Experimental Design

Statistical Analysis

BLE Protects AML12 Cells against Oxidative Stress-Induced Cytotoxicity

Conclusions