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


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.

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Wang, D. , Ishikawa, Y. , Miyazaki, M. , Fujita, H. , Tsutsui, K. , Sano, K. , Masuoka, N. and Ogino, K. (2011) A new risk assessment method for evaluation of oxidative chemicals using catalase mutant mouse primary hepatocytes. Health, 3, 288-291. doi: 10.4236/health.2011.35050.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Andersen, M.E. and Krewski, D. (2009) Toxicity tesing in the 21st century: Bringing the vision to life. Toxicological Sciences, 107, 324-330. doi:10.1093/toxsci/kfn255
[2] Halliwell, B. and Gutteridge, J.M.C. (2007) Cellular responses to oxidative stress: adaptation, damage, repair, senescence and death. In: Halliwell, B., Gutteridge, J.M.C., eds., Free Radicals in Biology and Medicine, 4rd Edition, Oxford University Press, New York, pp.187-267.
[3] Cohen, G. and Hochstein, P. (1963) Glutathione peroxidase: The primary agent for the elimination of hydrogen peroxide in erythrocytes. Biochemistry, 2, 1420-1428. doi:10.1021/bi00906a038
[4] Scott, M.D., Lubin, B.H., Zuo, L., Kuypers, F.A. (1991) Erythrocyte defense against hydrogen peroxide: preeminent importance of catalase. The Journal of Laboratory and Clinical Medicine, 118, 7-16.
[5] Boveris, A. and Chance, B. (1973) The mitochondrial generation hydrogen peroxide. Biochemical Journal, 134, 707-716.
[6] Feinstein, R.N., Howard, J.B., Braun, J.T. and Seaholm, J.E. (1966) Acatalasemic and hypocatalasemic mouse mutants. Genetics, 53, 923-933.
[7] Shaffer, J.B. and Preston, K.E. (1990) Molecular analysis of an acatalasemic mousemutant. Biochemical and Biophysical Research Communications, 173, 1043-1050. doi:10.1016/S0006-291X(05)80891-5
[8] Wang, D-H., Tsutsui, K., Sano, K., Masuoka, N. and Kira, S. (2001) cDNA cloning and expression of mutant catalase from the hypocatalasemic mouse: comparison with the acatalasemic mutant. Biochimica et Biophysica Acta, 1522, 217-220.
[9] Miyazaki, M. (1977) Primary culture of adult rat liver cells. I. Preparation of isolated cells from trypsin-per- fused liver of adult rat. Acta Medica Okayama, 31, 351- 360.
[10] Kondo, A., Miyazaki, M., Pu, H., Gao, C. and Namba, M. (1999) Establishment and cellular characteristics of a hepatocyte cell line (OUMS-31) derived from an acatalasemic mouse. In Vitro Cellular & Developmental Biology - Animal, 35, 155-158. doi:10.1007/s11626-999-0018-4
[11] Block, G.D., Locker, J., Bowen, W.C., Petersen, B.E., Katyal, S., Strom, S.C., Riley, T., Howard, T.A. and Michalopoulos, G.K. (1996) Population expansion, clonal growth, and specific differentiation patterns in primary cultures of hepatocytes induced by HGF/SF, EGF and TGF alpha in a chemically defined (HGM) medium. The Journal of Cell Biology, 132, 1133-1149. doi:10.1083/jcb.132.6.1133
[12] Miyazaki, M., Akiyama, I., Sakaguchi, M., Naka Shima, E., Okada, M., Kataoka, K. and Huh, N.H. (2002) Improved conditions to induce hepatocytes from rat bone marrow cells in culture. Biochemical and Biophysical Research Communications, 298, 24-30. doi:10.1016/S0006-291X(02)02340-9
[13] Wang, D-H., Tsutsui, K. and Kira, S. (2001) Detecting genotypes of catalase mutant mice by genomic polymerase chain reaction and restriction analysis. Analytical Biochemistry, 299, 116-117. doi:10.1006/abio.2001.5356
[14] Tominaga, H., Ishiyama, M., Ohseto, F., Sasamoto, K., Hamamoto, T., Suzuki K. and Watanabe, M. (1999) A water-soluble tetrazolium salt useful for colorimetric cell viability assay. Annals of Communications, 36, 47-50. doi:10.1039/a809656b
[15] Scientific Committee on Cosmetic Products and Non-Food Products Intended for Consumers (2004) Opinion concerning lawsone. Colipa No. C146. SCCNFP/0798/04, February 16.
[16] Wang, D-H., Masuoka, N. and Kira, S. (2003) Animal model for oxidative stress research - catalase mutant mice. Environmental Health and Preventive Medicine, 8, 37-40. doi:10.1007/BF02897924
[17] Sauriasari, R., Wang, D-H., Takemura, Y., Tsutsui, K., Masuoka, N., Sano, K., Horita, M., Wang, B.L. and Ogino, K. (2007) Cytotoxicity of lawsone and cytoprotective activity of antioxidants in catalase mutant. Escherichia Coli. Toxicology, 235, 103-111. doi:10.1016/j.tox.2007.03.019
[18] Miyazaki, M. (1978) Primary culture of adult rat liver cells. II. Cytological and biochemical properties of primary cultured cells. Acta Medica Okayama, 32, 11-22.
[19] Miyazaki, M., Watanabe, A., Shiraishi, M., Hoshika, T., Miyano, K. and Sato, J. (1978) Primary culture of adult rat liver cells. III. Hormonal effects on cytological and biochemical properties of primary cultured cells. Acta Medica Okayama, 32, 85-96.

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