Metabolism of ethanol to acetaldehyde and increased susceptibility to oxidative stress could play a role in the ovarian tissue cell injury promoted by alcohol drinking
Restricted accessResearch articleFirst published online September, 2009
Metabolism of ethanol to acetaldehyde and increased susceptibility to oxidative stress could play a role in the ovarian tissue cell injury promoted by alcohol drinking
It is known that drinking alcohol can lead to reproductive problems in women. In this study, we analyzed the possibility that part of those effects were mediated through alterations of ovarian function related to ethanol oxidation to acetaldehyde occurring in situ. Biotransformation in the rat ovary cytosolic fraction was partially inhibited by allopurinol, suggesting the participation of xanthine oxidoreductase in the process. Microsomal pathway was of enzymatic nature, requiring nicotinamide adenine dinucleotide phosphate-oxidase (NADPH), sensitive to oxygen and significantly inhibited by sodium diethyldithiocarbamate, 4-methylpyrazole and diphenyleneiodonium. Aldehyde dehydrogenase activity was detected by histochemistry in the ovarian tissue, in the strome surrounding the follicle while no alcohol dehydrogenase was detected. However, biochemical determination of alcohol dehydrogenase and aldehyde dehydrogenase activities in rat ovarian tissue revealed the presence of some activity of both enzymes but significantly lower than those found in the liver. By repetitive exposure of animals to ethanol, the microsomal metabolism to acetaldehyde was increased but not in the case of the cytosolic fraction. In these animals, t-butylhydroperoxyde-promoted chemiluminiscence was increased in comparison to control, revealing an increased susceptibility to oxidative stress due to alcohol drinking. Ultrastructure of ovarian tissue from rats exposed chronically to alcohol revealed alterations at the level of the granulosa; theca interna and pellucida zones. In the secondary follicle, alterations consisted of marked condensation of chromatin attached to the nuclear inner membrane. Intense dilatation of the outer perinuclear space could be observed. There was a marked dilatation of the rough endoplasmic reticulum accompanied of significant detachment of ribosomes from their membranes. Mitochondria appeared swollen. In the zona pellucida, most of the cell processes from oocyte and corona radiata cells were absent or broken totally or in part. Results suggest that in the rat ovary, metabolism of ethanol to acetaldehyde may play a role in alcohol effects on female reproductive function.
Aleynik SI, Leo MA, Aleynik MK, and Lieber CS ( 1999) Alcohol-induced pancreatic oxidative stress: protection by phospholipid repletion. Free Radical Biology and Medicine26: 609-619.
2.
Allali-Hassani A., Martinez SE, Peralba JM, et al. (1997) Alcohol dehydrogenase of human and rat blood vessels. Role in ethanol metabolism. FEBS Letters405: 26-30.
3.
Boveris A., Fraga CG, Varsavsky AI, and Koch OR ( 1983) Increased chemiluminiscence and superoxide production in liver of chronically ethanol treated rats. Archives of Biochemistry and Biophysics227: 537-541.
4.
Campillo B. ( 2005) Hormonal responses in alcoholism. In: Watson RR and Preedy V (eds.) Comprehensive Handbook of Alcohol Related Pathology, Vol. 2. London: Academic Press, pp.982-983.
5.
Castro JA, Castro GD ( 2005) Mechanisms in prostate damage by alcohol. In: Watson RR and Preedy V (eds.) Comprehensive Handbook of Alcohol Related Pathology, vol. 2. London: Academic Press, pp.1007-1015.
6.
Castro GD, Delgado de Layño AM, Costantini MH, and Castro JA ( 2001) Cytosolic xanthine oxidoreductase mediated bioactivation of ethanol to acetaldehyde and free radicals in rat breast tissue. Its potential role in alcohol-promoted mammary cancer. Toxicology160: 11-18.
7.
Castro GD, Delgado de Layño AM, Costantini MH, and Castro JA ( 2003) Rat breast microsomal biotransformation of ethanol to acetaldehyde but not to free radicals: Its potential role in the association between alcohol drinking and breast tumor promotion. Teratogenesis, Carcinogenesis, and Mutagenesis23: 61-70.
8.
Castro GD, Delgado de Layño AM, Fanelli SL, Maciel ME, Díaz Gómez MI, and Castro JA ( 2008) Acetaldehyde accumulation in rat mammary tissue after an acute treatment with alcohol. Journal of Applied Toxicology28: 315-321.
9.
Castro GD, Rodríguez de Castro C., Maciel ME, et al. (2006) Ethanol-induced oxidative stress and acetaldehyde formation in rat mammary tissue: potential factors involved in alcohol drinking promotion of breast cancer. Toxicology210: 208-219.
10.
Davis BJ, Heindel JJ ( 1998) Ovarian toxicants: multiple mechanisms of action . In: Korach KS (ed.) Reproductive and Developmental Toxicology. New York: Marcel Dekker, pp.373-396.
11.
de Castro CR, de Toranzo EG, Bernacchi AS, Carbone M., and Castro JA ( 1989) Ultrastructural alterations in ovaries from nifurtimox or benznidazole-treated rats: their relations to ovarian nitroreductive biotransformation of both drugs. Experimental and Molecular Pathology50: 385-397.
12.
Dellarco VL ( 1998) A mutagenicity assessment of acetaldehyde. Mutation Research195: 1-20.
13.
Díaz Gómez MI, Castro GD, Delgado de Layño AM, Costantini MH, and Castro JA ( 2000) Cytochrome P450 reductase mediated anaerobic biotransformation of ethanol to 1-hydroxyethyl free radicals and acetaldehyde. Toxicology154: 113-122.
14.
Díaz Gómez MI, Fanelli SL, Delgado de Layño AM, Castro JA, and Castro GD ( 2006) Liver nuclear and microsomal CYP2E1-mediated metabolism of xenobiotics in rats chronically drinking an alcohol-containing liquid diet . Toxicology and Industrial Health22: 367-374.
15.
Díaz Gómez MI, Fanelli SL, Delgado de Layño AM, Bietto FM, Castro JA, and Castro GD ( 2008) Deleterious effects induced by oxidative stress in liver nuclei from rats receiving an alcohol-containing liquid diet. Toxicology and Industrial Health24: 625-634.
16.
Díaz Gómez MI, Tamayo D., and Castro JA ( 1988) Nitrosodimethylamine metabolism in rat ovaries. Interactions of its metabolites with nucleic acids and proteins. Cancer Letters41: 257-263.
17.
Egerer G., Stickel F., and Seitz HK ( 2005) Alcohol and the gastrointestinal tract. In: Watson RR and Preedy V (eds.) Comprehensive Handbook of Alcohol Related Pathology, vol 2. London: Academic Press, pp.557-569.
18.
Emanuele N., Emanuele MA ( 1997) The endocrine system: alcohol alters critical hormonal balance. Alcohol Health and Research World21: 53-64.
19.
Frederiks WM, Bosch KS, Van den Munckhof RJ, and Van Noorden CJ ( 1994) A quantitative histochemical study of xanthine oxidase activity in rat liver using the cerium capture method in the presence of polyvinyl alcohol. The Journal of Histochemistry and Cytochemistry42: 1091-1097.
20.
Gad SC ( 2001) Statistics for toxicologists. In: Hayes AW (ed.) Principles and Methods of Toxicology. 4th ed. Philadelphia , Taylor & Francis pp.285-364.
21.
Galvaõ-Teles A., Monteiro E., Gavaler JS, and Van Thiel DH ( 1986) Gonadal consequences of alcohol abuse: lessons from the liver. Hepatology6: 135-140.
22.
Garro AJ, Gordon BHJ, and Lieber CS ( 1992) Alcohol abuse: carcinogenic effects and fetal alcohol syndrome. Description, diagnosis and prevention. In: Lieber CS (ed.) Medical and Nutritional Complications of Alcoholism. Mechanisms and Management. New York: Plenum Publishing, pp.459-493.
23.
Gavaler JS, Van Thiel DH ( 1987) Reproductive consequences of alcohol abuse: males and females compared and contrasted. Mutation Research186: 269-277.
24.
Kapoor N., Pant AB, Dhawan A., et al. (2006) Differences in sensitivity of cultured rat brain neuronal and glial cytochrome P450 2E1 to ethanol. Life Sciences79: 1514-1522.
25.
Keating AF, Rajapaksa KS, Sipes IG, and Hoyer PB ( 2008) Effect of CYP2E1 gene deletion in mice on expression of microsomal epoxy hydrolase in response to VCD exposure. Toxicological Sciences105: 351-359.
26.
Koivula T., Koivusalo M. ( 1975) Different forms of rat liver aldehyde dehydrogenase and their subcellular distribution. Biochimica et Biophysica Acta397: 9-23.
27.
Kulkarni AP ( 2002) Lipoxygenases. In: Ioannides C (ed.) Enzyme systems that metabolise drugs and other xenobiotics . New York: John Wiley and Sons , pp.231-279.
28.
Lieber CS ( 2005) alcohol metabolism: general aspects. In: Watson RR and Preedy V (eds.) Comprehensive Handbook of Alcohol Related Pathology, vol 1. London: Academic Press, pp.1211-1222.
29.
Lieber CS, De Carli LM ( 1982) The feeding of alcohol in liquid diets: two decades of applications and 1982 update. Alcoholism Clinical and Experimental Research6: 523-531.
30.
Lieber CS, De Carli LM ( 1989) Liquid diet technique of ethanol administration, 1989 update. Alcohol and Alcoholism24: 197-211.
31.
Lowry OH, Rosebrough NJ, Farr AL, and Randall RJ ( 1951) Protein measurement with the Folin phenol reagent . The Journal of Biological Chemistry193: 265-275.
32.
Lu Y., Cederbaum AI ( 2008) CYP2E1 and oxidative liver injury by alcohol. Free Radical Biology and Medicine44: 723-738.
33.
Maciel ME, Castro GD, and Castro JA ( 2004) Inhibition of the rat breast cytosolic bioactivation of ethanol to acetaldehyde by some plant polyphenols and folic acid. Nutrition and Cancer49: 94-99.
34.
Mattisson DR ( 1985) Clinical manifestations of ovarian toxicity. In: Dixan RC (ed.) Reproductive Toxicology . New York: Raven Press , pp.109-130.
35.
Mattison DR, Plowchalk DR, Meadows MJ, al-Juburi AZ, Gandy J., and Malek A. ( 1990) Reproductive toxicity: male and female reproductive systems as targets for chemical injury. The Medical Clinics of North America74: 391-411.
36.
McGuire JJ, Anderson DJ, McDonald BJ, Narayanasami R., and Bennett BM ( 1998) Inhibition of NADPH-cytochrome P450 reductase and glyceryl trinitrate biotransformation by diphenyleneiodonium sulfate. Biochemical Pharmacology56: 881-893.
37.
Mello NK, Mendelson JH, and Teak SK ( 1993) An overview of the effects of alcohol on neuroendocrine function in women. In: Zakhari S (ed) Alcohol and the Endocrine System. National Institute on Alcohol Abuse and Alcoholism Research Monograph N° 23, NIH Pub. N° 93-3533, Bethesda, MD: The Institute, pp.139-169.
38.
Mendelson JH, Mello NK ( 1988) Chronic alcohol effects on anterior pituitary and ovarian hormones in healthy women. Journal of Pharmacology and Experimental Therapeutics245: 407-412.
39.
Norman AW, Litwick G. ( 1987) Hormones. 2nd ed. New York: Academic Press, pp.361-365.
40.
Opsian DD, Coon MJ ( 1982) Oxidation-reduction states of FMN and FAD in NADPH-cytochrome P450 reductase during reduction of NADPH. Journal of Biological Chemistry257: 8935-8944.
41.
Quintans LN, Castro GD, and Castro JA ( 2005) Oxidation of ethanol to acetaldehyde and free radicals by rat testicular microsomes. Archives of Toxicology79: 25-30.
42.
Racker E. ( 1950) Crystalline alcohol dehydrogenase from baker’s yeast . The Journal of Biological Chemistry184: 313-319.
43.
Rendic S., Di Carlo FJ ( 1997) Human cytochrome P450 enzymes: a status report summarizing their reactions, substrates, inducers, and inhibitors. Drug Metabolism Reviews29: 413-580.
44.
Rhodin JAG ( 1974) Histology. A Text and Atlas. New York : Oxford University Press, pp.704-718.
45.
Saxena S., Meehan D., Coney P., and Wimalasena J. ( 1990) Ethanol has a direct inhibitory effects on steroideogenesis in human granulosa cells: specific inhibition of LH action. Alcoholism Clinical and Experimental Research14: 522-527.
46.
Shibley IA, Jr, McIntyre TA, and Pennington SN ( 1999) Experimental models used to measure direct and indirect ethanol teratogenicity. Alcohol and Alcoholism34: 125-140.
47.
Smith BJ, Mattison DR, and Sipes IG ( 1990) The role of epoxidation in 4-vinylcyclohexane-induced ovarian toxicity. Toxicology and Applied Pharmacology105: 372-381.
48.
Törok B. ( 2004) Assessing problems of organic hydroperoxide induced chemiluminescence related to biological/ medical samples. Journal of Biochemical and Biophysical Methods61: 247-251.
49.
Villarruel MC, de Toranzo EGD, Castro JA ( 1977) CCl4 activation, lipid peroxidation and the mixed function oxygenase activity of various rat tissues. Toxicology and Applied Pharmacology41: 337-344.
