A new risk assessment method for evaluation of oxidative chemicals using catalase mutant mouse primary hepatocytes

Da-Hong Wang,Hirofumi Fujita,Yasuo Ishikawa,Kuniaki Sano,Ken Tsutsui,Masahiro Miyazaki,Keiki Ogino,Noriyoshi Masuoka

doi:10.4236/health.2011.35050

Abstract

We examined the possibility of developing a new risk assessment method for potentially hazardous chemicals by using mouse primary hepatocytes from acatalasemic mice (Csb) and the wild-type (Csa) as predictive model. Chemical-induced cytotoxicities, such as hydrogen peroxide and lawsone, a main hair dye ingredient of henna, were examined. We observed the differences in cell survival between the Csa and Csb in a dose-dependant manner after treatment with either hydrogen peroxide or lawsone, supporting the usefulness of this newly established method for hazard identification of oxidative chemicals in a risk assessment process. More chemicals will be tested to confirm the usefulness of this method for the preliminary screening of oxidative chemicals before animal experimentation.

Highlights

More and more chemicals are synthesized for industrial and consumer use, it is impossible to finish long-term rodent bioassay for detection of carcinogens and identification of hazards in all chemicals because it involves large numbers of animals and is extremely expensive
We examined the possibility of developing a new risk assessment method for evaluation of oxidative chemicals using mouse primary hepatocytes from acatalasemic mice (Csb) and the wild-type (Csa) as predictive model
Based on the cell viability, we can use the Csa and Csb as predictive models to assume whether H2O2 involves in the chemical-induced cytotoxicity, which would be helpful for the preliminary screening of the potential oxidative chemicals

Summary

INTRODUCTION

More and more chemicals are synthesized for industrial and consumer use, it is impossible to finish long-term rodent bioassay for detection of carcinogens and identification of hazards in all chemicals because it involves large numbers of animals and is extremely expensive. Simple and efficient pre-screening alternatives to animal experimentation are desirable [1]. Catalase-mutant acatalasemic mouse was established by Feinstein et al through a large scale screening of the progeny of irradiated C3H mice [6]. A point mutation at amino acid 11 (from glutamine to histidine) of Csb mouse catalase is responsible for its catalase deficiency [7,8]. We examined the possibility of developing a new risk assessment method for evaluation of oxidative chemicals using mouse primary hepatocytes from acatalasemic mice (Csb) and the wild-type (Csa) as predictive model

Hepatocyte Isolation

Confirmation of the Genotype of Catalase Mutant Mouse Hepatocytes

Cell Viability Assay

The Csb Catalase Mutant Gene Is Susceptible to NdeI Digestion

Comparison of Lawsone-Induced

CONCLUSIONS