Hydrogen peroxide induces acute injury and up-regulates inflammatory gene expression in hepatocytes: An in vitro model

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

In the past, many in vitro hepatocyte injury models developed for liver regeneration used carbon tetrachloride as irritant chemical. Recently, carbon tetrachloride usage was prohibited due to serious deleterious effects to human and environment. There is an urgent need to develop a new acute chemical-induced hepatocyte injury model using other chemical compound to replace carbon tetrachloride. In this study, we used hydrogen peroxide (H2O2) to induced hepatocyte injury with HepG2 as the liver cell model. HepG2 injury was established by exposing the cells to CC50 of H2O2 at the concentration of 2.4 mM, predetermined via MTT assay for 2 h exposure. Aspartate aminotransferase (AST) activity was measured to determine the extent of cellular injury and quantitative PCR was carried out to determine the expression of inflammatory genes of the cells 24 h after H2O2 exposure. The results showed that AST activity increased with time and peak at 24 h after H2O2 exposure. Quantitative PCR analysis demonstrated that expression of inflammatory genes (TGF-β1, MMP-3, NF-xβ, IL-8 and IL-6) increased significantly. In addition, the gene expression of GPX, an anti-oxidant enzyme was also increased significantly in response to oxidative stress. In summary, H2O2 demonstrated excellent capability in inducing oxidative injury to HepG2 and together they represent an ideal acute chemical-induced injury model that can be used for liver regeneration study. Our results also provide input for inflammatory gene expression in the hepatocyte injury model.

Original languageEnglish
Pages (from-to)451-458
Number of pages8
JournalSains Malaysiana
Volume45
Issue number3
Publication statusPublished - 1 Mar 2016

Fingerprint

Hydrogen Peroxide
Hepatocytes
Up-Regulation
Gene Expression
Wounds and Injuries
Carbon Tetrachloride
Liver Regeneration
Aspartate Aminotransferases
Polymerase Chain Reaction
In Vitro Techniques
Irritants
Matrix Metalloproteinases
Interleukin-8
Oxidants
Interleukin-6
Oxidative Stress
Liver
Enzymes

Keywords

  • HO
  • Hepatocytes
  • In vitro
  • Inflammatory genes
  • Liver injury model

ASJC Scopus subject areas

  • General

Cite this

@article{401eaa9f224b4b85815b363c51ed4971,
title = "Hydrogen peroxide induces acute injury and up-regulates inflammatory gene expression in hepatocytes: An in vitro model",
abstract = "In the past, many in vitro hepatocyte injury models developed for liver regeneration used carbon tetrachloride as irritant chemical. Recently, carbon tetrachloride usage was prohibited due to serious deleterious effects to human and environment. There is an urgent need to develop a new acute chemical-induced hepatocyte injury model using other chemical compound to replace carbon tetrachloride. In this study, we used hydrogen peroxide (H2O2) to induced hepatocyte injury with HepG2 as the liver cell model. HepG2 injury was established by exposing the cells to CC50 of H2O2 at the concentration of 2.4 mM, predetermined via MTT assay for 2 h exposure. Aspartate aminotransferase (AST) activity was measured to determine the extent of cellular injury and quantitative PCR was carried out to determine the expression of inflammatory genes of the cells 24 h after H2O2 exposure. The results showed that AST activity increased with time and peak at 24 h after H2O2 exposure. Quantitative PCR analysis demonstrated that expression of inflammatory genes (TGF-β1, MMP-3, NF-xβ, IL-8 and IL-6) increased significantly. In addition, the gene expression of GPX, an anti-oxidant enzyme was also increased significantly in response to oxidative stress. In summary, H2O2 demonstrated excellent capability in inducing oxidative injury to HepG2 and together they represent an ideal acute chemical-induced injury model that can be used for liver regeneration study. Our results also provide input for inflammatory gene expression in the hepatocyte injury model.",
keywords = "HO, Hepatocytes, In vitro, Inflammatory genes, Liver injury model",
author = "Liau, {Ling Ling} and {Kien Hui}, Chua and Suzana Makpol and {Abdul Ghani}, {Nur Azurah}",
year = "2016",
month = "3",
day = "1",
language = "English",
volume = "45",
pages = "451--458",
journal = "Sains Malaysiana",
issn = "0126-6039",
publisher = "Penerbit Universiti Kebangsaan Malaysia",
number = "3",

}

TY - JOUR

T1 - Hydrogen peroxide induces acute injury and up-regulates inflammatory gene expression in hepatocytes

T2 - An in vitro model

AU - Liau, Ling Ling

AU - Kien Hui, Chua

AU - Makpol, Suzana

AU - Abdul Ghani, Nur Azurah

PY - 2016/3/1

Y1 - 2016/3/1

N2 - In the past, many in vitro hepatocyte injury models developed for liver regeneration used carbon tetrachloride as irritant chemical. Recently, carbon tetrachloride usage was prohibited due to serious deleterious effects to human and environment. There is an urgent need to develop a new acute chemical-induced hepatocyte injury model using other chemical compound to replace carbon tetrachloride. In this study, we used hydrogen peroxide (H2O2) to induced hepatocyte injury with HepG2 as the liver cell model. HepG2 injury was established by exposing the cells to CC50 of H2O2 at the concentration of 2.4 mM, predetermined via MTT assay for 2 h exposure. Aspartate aminotransferase (AST) activity was measured to determine the extent of cellular injury and quantitative PCR was carried out to determine the expression of inflammatory genes of the cells 24 h after H2O2 exposure. The results showed that AST activity increased with time and peak at 24 h after H2O2 exposure. Quantitative PCR analysis demonstrated that expression of inflammatory genes (TGF-β1, MMP-3, NF-xβ, IL-8 and IL-6) increased significantly. In addition, the gene expression of GPX, an anti-oxidant enzyme was also increased significantly in response to oxidative stress. In summary, H2O2 demonstrated excellent capability in inducing oxidative injury to HepG2 and together they represent an ideal acute chemical-induced injury model that can be used for liver regeneration study. Our results also provide input for inflammatory gene expression in the hepatocyte injury model.

AB - In the past, many in vitro hepatocyte injury models developed for liver regeneration used carbon tetrachloride as irritant chemical. Recently, carbon tetrachloride usage was prohibited due to serious deleterious effects to human and environment. There is an urgent need to develop a new acute chemical-induced hepatocyte injury model using other chemical compound to replace carbon tetrachloride. In this study, we used hydrogen peroxide (H2O2) to induced hepatocyte injury with HepG2 as the liver cell model. HepG2 injury was established by exposing the cells to CC50 of H2O2 at the concentration of 2.4 mM, predetermined via MTT assay for 2 h exposure. Aspartate aminotransferase (AST) activity was measured to determine the extent of cellular injury and quantitative PCR was carried out to determine the expression of inflammatory genes of the cells 24 h after H2O2 exposure. The results showed that AST activity increased with time and peak at 24 h after H2O2 exposure. Quantitative PCR analysis demonstrated that expression of inflammatory genes (TGF-β1, MMP-3, NF-xβ, IL-8 and IL-6) increased significantly. In addition, the gene expression of GPX, an anti-oxidant enzyme was also increased significantly in response to oxidative stress. In summary, H2O2 demonstrated excellent capability in inducing oxidative injury to HepG2 and together they represent an ideal acute chemical-induced injury model that can be used for liver regeneration study. Our results also provide input for inflammatory gene expression in the hepatocyte injury model.

KW - HO

KW - Hepatocytes

KW - In vitro

KW - Inflammatory genes

KW - Liver injury model

UR - http://www.scopus.com/inward/record.url?scp=84964328353&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84964328353&partnerID=8YFLogxK

M3 - Article

AN - SCOPUS:84964328353

VL - 45

SP - 451

EP - 458

JO - Sains Malaysiana

JF - Sains Malaysiana

SN - 0126-6039

IS - 3

ER -