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ToxSci Advance Access published online on April 15, 2003
Toxicological Sciences, doi:10.1093/toxsci/kfg082
Toxicological Sciences © Society of Toxicology 2003; all rights reserved
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Received November 23, 2002; accepted March 6, 2003
© 2003 Society of Toxicology

Molecular and Genetic Toxicology

The Effect of Cigarette Smoke Exposure and Ascorbic Acid Intake on Gene Expression of Antioxidant Enzymes and Other Related Enzymes in the Livers and Lungs of Osteogenic Disorder Shionogi Rats

Etsuko Ueta 1, Yuko Tadokoro 1, Tomoko Yamamoto 2, Chiyuki Yamane 2, Emiko Suzuki 3, Eiji Nanba 4, Yuzuru Otsuka 1*, Tadao Kurata 1

1 Institute of Environmental Science for Human Life, Ochanomizu University, Tokyo, 112-8610, Japan
2 Faculty of Education and Regional Sciences, Tottori University, Tottori, 680-8551, Japan
3 Faculty of Life Science, Ochanomizu University, Tokyo, 112-8610, Japan
4 Gene Research Center, Tottori University, Yonago, 683-0826, Japan

* To whom correspondence should be addressed. E-mail: yotsuka{at}cc.ocha.ac.jp.


  Abstract

Cigarette smoking causes many chronic diseases but is a preventable risk factor in developing countries. However, it may be possible to relieve the smoke induced damage by increasing the protective defense system. As vitamin C intake reduces smoking risk, it is recommended that smokers should take more vitamin C. However, the molecular mechanism of vitamin C intake on smokers has not been thoroughly investigated. We have found there to be suppression of smoke induced cytochrome P-450 1A1 (CYP1A1) mRNA expression by high dose ascorbic acid administration. Therefore, we surveyed other genes the expressions of which were altered by the administration of high dose ascorbic acid. As cigarette smoking increases oxidative stress, we investigated the effect on anti-oxidative enzymes expression. The Osteogenic Disorder Shionogi (ODS) rat which lacks ascorbic acid synthesis enzyme, were administered either minimal amounts (4 mg/day, S4) or high dose amounts (40 mg/day, S40) of ascorbic acid, and were exposed to cigarette smoke daily for 25 days. The effect on anti-oxidative enzymes mRNA expression in the liver was measured by competitive reverse transcription - polymerase chain reaction method (competitive RT-PCR). CuZn-superoxide dismutase (SOD), MnSOD, catalase and protein disulfide isomerase (PDI) were significantly decreased by high dose ascorbic acid administration, and plasma glutathione peroxidase was also decreased but not significantly. Cigarette smoke exposure slightly increased gene expression of PDI and catalase, but not significantly. The differently expressed 27 genes in the liver were found by differential display methods. From 27 genes, altered expression of plasma proteinase inhibitor alpha-1-inhibitor III and CYP1A2 were confirmed by competitive RT-PCR. These results show that ascorbic acid intake influences gene expression of anti-oxidative enzymes, an ascorbic acid recycle enzyme and xenobiotic metabolizing enzymes.

Ascorbic acid, Cigarette smoke, Gene expression, Differential display, Competitive RT-PCR, CYP1A2, Plasma proteinase inhibitor alpha-1-inhibitor III .


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