Journal of Basic and Applied Sciences Vol. 4, No.

2, 105-108, 2008

ISSN: 1814-8085

EFFECT OF VITAMIN C ON MALONDIALDEHYDE (MDA) IN PREGNANT NIGERIAN WOMEN

Luqman A. Olayaki1*, Salihu M. Ajao2, Gafar A.A. Jimoh3, I.T. Aremu1 and Ayodele O. Soladoye1 1 Department of Physiology, 2Department of Anatomy, 3Department Obstetrics and Gynecology College of Health Sciences, University of Ilorin, Ilorin, Nigeria ABSTRACT The experiment was carried out to evaluate the effects of vitamin C on lipid peroxidation and non-enzymatic antioxidant status during second trimester in normal pregnancy in Nigerians. The study comprised 60 age-matched subjects divided into 3 groups of a) 20 healthy non-pregnant women, b) 20 pregnant women and c) 20 pregnant women with vitamin C treatment. Estimation of lipid peroxidation was done by thiobarbituric reactive substance (TBARS) by standard method. Data analysis was by repeated-measures analysis of variance, using SPSS software (version 11.0; SPSS Inc, Chicago IL). Statistically significant increase in levels of lipid peroxidation antioxidants were observed in pregnant women as compared to levels in non-pregnant women and vitamin C treatment significantly reduced lipid peroxidation during pregnancy. Normal pregnancy is associated with increased malondialdehyde production, and ingestion of vitamin C reduced its production. Keywords: Pregnancy, malondialdehyde, vitamin c, second trimester. INTRODUCTION Normal pregnancy is accompanied by a high metabolic demand and elevated requirements for tissue oxygen, which results in increased oxidative stress and antioxidant defenses (Knapen, 1999). It is well established that lipid peroxides, such as thiobarbituric acid reactive substances increase significantly in the maternal circulation (Arikan et al., 2001). Morris et al (1998) reported high circulating levels of lipid peroxides in normal pregnancy and preeclampsia. Pregnancy also has an effect on maternal antioxidant enzyme activities. Glutathione peroxidase and superoxide dismutase activities have been found to be reduced during the second trimester of pregnancy in humans (Zachara et al., 1993; Qanungo and Mukherjea, 2000). Malondialdehyde (MDA) is a decomposition product of peroxidised polyunsaturated fatty acids (Mukai and Goldstein, 1976) that is widely preferred for detection of free oxygen radicals in various pathological conditions (Lazzarino et al., 1994). Its level has been shown to increase with gestational age (Patil et al., 2006). One important pathological condition during pregnancy is pre-eclampsia. This is a leading cause of premature delivery and foetal growth retardation (Lim and Friedman, 1993). Pre-eclampsia is associated with increased lipid peroxidation in the maternal circulation and in the placenta (Walsh and Wang, 1998). Vitamin C is a hydrophilic molecule that can scavenge several radicals, among them the hydroxyl radical (Cederberg et al., 2001). Its level is known to decrease during pregnancy (FNB, 2000). In view of the antioxidant roles of vitamin C, we decided to investigate its effects on malondialdehyde (MDA) in pregnant Nigerian women during the second trimester. SUBJECTS AND METHODS The subjects enrolled in this study comprised of 40 healthy pregnant women in the second trimester within the age range of 20-35 years, they were divided into two groups of twenty pregnant women each (groups II and III). Twenty age and sex-matched healthy non-pregnant adult females enrolled for comparative studies (group I). Exclusion criteria include obesity, smoking, diabetes mellitus, alcoholism, severe anemia (Hb < 6.0 g/dL), and any other systemic disorder. None of the study or control subjects had taken any form of vitamin C supplementation within a period of one month prior to participation in the study. Each of the subjects in group III took 100mg of vitamin C daily. Two blood samples were collected from each of the pregnant women within six weeks during the second trimester, and one blood sample was collected from each of the non-pregnant women. Fasting (8h) blood samples were collected at entry to the study in early second trimester and six weeks later from study participants. Five mL of blood was drawn by

*Corresponding author. E-mail: luqmanolayaki@yahoo.com

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J. basic appl. sci. Several studies have indicated that the antioxidative defense system is modified during pregnancy. Wisdom et al. (1991) showed that the activity of an important family of antioxidative enzymes, the superoxide dismutase (SOD), is reduced in the blood of pregnant women. In addition, Walsh and Wang (1993b) reported a deficiency in another antioxidative enzyme glutathione peroxidase (GPx) during pregnancy. GPx is an important antioxidant enzyme present in virtually all tissues. The enzyme limits the generation of lipid peroxides and utilizes glutathione as its cofactor to convert lipid peroxides into relatively harmless hydroxylated fatty acids, water and glutathione disulfide (Levander, 1992). Given these findings, it might be expected that a deficiency in these enzyme would lead to increased lipid peroxidation during pregnancy. Serum level of vitamin C and vitamin E have been observed to be reduced throughout pregnancy (Kharb et al., 2000; Hassan and Onu, 2006). In the present study, the plasma concentration of malondialdehyde (MDA) is higher in pregnancy state than in non-pregnant state, and the concentration increases with progressing gestational age. This is in agreement with earlier study by Patil et al (2006). This could be due to increased generation of ROS because of increased oxygen demand during pregnancy (Halliwell and Gutteridge, 1990), reduction in the activities of antioxidant enzymes such as superoxide dismutase and glutathione peroxidase (Walsh and Wang, 1993b), and reduction in plasma vitamin C and vitamin E concentration during pregnancy (Kumar and Das, 2002; Kharb et al., 2000). Ingestion of vitamin C during pregnancy was observed to reduce the generation of MDA. Vitamin C is a primary water-soluble antioxidant in plasma and within cells, but it can also interact with the plasma membrane by donating electrons to the -tocopheroxyl radical and a trans-plasma membrane oxidoreductase activity. Vitamin C supplementation is particularly important in pregnant women as its deficiency has been shown to affect placental structure and facilitates placental infection both of which results in increased risk of premature rupture of placental membranes and premature births (Casanueva and Viteri, 2002). In addition the supplementation could help to prevent the development of such complications of pregnancy like gestational hypertension, intrauterine growth retardation and gestational diabetes, all known to be associated with high levels of oxidative stress, in addition to the numerous other benefits of this vitamin in human metabolism and health (Rumbold and Crowther, 2005). Lipid peroxidation is known to be potentially harmful because its uncontrolled, self-enhancing process causes disruption of membrane lipids and other cell components (Mahboob et al., 2005), this could have

venepuncture and collected in a heparinized tube (10 units of heparin per mL of blood). The malondialdehyde (MDA) was analyzed within 10-15 minutes of collecting blood sample in the whole blood by measurement of thiobarbituric acid reactive substance (TBARS). Data analysis was by repeated-measures analysis of variance, using SPSS software (version 11.0; SPSS Inc, Chicago. IL). ETHICS The study was conducted in compliance with the Declaration on the Rights of the Patient (WMA, 2000). RESULTS A comparison of malondialdehyde values measured within six weeks during second trimester of pregnancy in Nigerian women is shown in Table 1. and Fig.1. There was an increase in malondialdehyde concentration during pregnancy (Groups II and III) compared to non pregnant state (Group I), from 1.660.02 mmol/L in group I to 1.800.03 mmol/L and 1.81 mmol/L in groups II and III respectively (p< 0.005), during the first contact. There was further increase in the concentration of malondialdehyde as pregnancy progresses (six weeks later) compared to the initial measurement. In group II (untreated pregnant women), malondialdehyde concentration increased from 1.800.03 mmol/L at first contact to 1.94 mmol/L (p<0.005) six weeks after. In group III (vitamin C-treated women), there was reduction in the concentration of malondialdehyde after six weeks of vitamin C ingestion, from 1.810.03 mmol/L at first contact to 1.79 mmol/L after six weeks, which was not statistically significant (p=0.39). DISCUSSION This study shows that physiological state of pregnancy causes an increase in the amount of lipid peroxidation products in the blood. It also shows that ingestion of vitamin C causes reduction in the amount of products of lipid peroxidation produced. Normal pregnancy is accompanied by a high metabolic demand and elevated requirements for tissue oxygen, which results in increased oxidative stress and antioxidant defenses (Knapen et al., 1999). Arikan et al (2001) reported significant increase in the level of thiobarbituric acid during normal pregnancy. Lipid peroxides are formed when lipids interact with oxygen radicals. The human placenta produces lipid peroxides that are secreted mainly to the maternal side of the placenta (Walsh and Wang, 1993a) and makers of increased lipid peroxidation are observed during normal pregnancy (Morris et al., 1998).

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Table 1. Malondialdehyde (MDA) Changes in Group I (Non-Pregnant), Group II (Untreated Pregnancy), and Group III (Vitamin C-Treated Pregnancy). Groups Control Untreated pregnancy (1st Day) Untreated pregnancy (After 6 weeks) Pregnancy + Vit C (1st Day) Pregnancy + Vit C (After 6 weeks) MDA (mmol/L) 1.660.02a 1.800.03 1.940.03b 1.810.03 1.790.03

Group I Group II Group III

Values are mean SEM, n=20, a p<0.001 versus others, b p<0.005 versus others.
2.5 2
b a

MDA (mmol/L)

1.5 1 0.5 0

CNTRL

PREGa

PREGb

PREG+VITCa

PREG+VITCb

Fig. 1: MeanSEM Malondialdehyde (MDA) in the plasma of non-pregnant (Control), pregnant (PREGa and PREGb) at 1st and 2nd contact, pregnant with vitamin C (PREG+VITCa and PREG+VITCb) women at 1st and 2nd contact. n=20 subjects per group, ap<0.005 versus other groups; b p<0.005 versus other groups. undesirable effects on foetal development and maternal health. In conclusion we have demonstrated that normal pregnancy is associated with increased MDA production. The positive influences of vitamin C on MDA production suggest a potential need for vitamin C during pregnancy in order to reduce lipid peroxidation. There may be need for vitamin C to be included among the routine antenatal drugs. REFERENCES Arikan, S. Konukoglu D. Arikan C. Akcay T. and Davas I. 2001. Lipid peroxidation and antioxidant status in maternal and cord blood. Gynecol Obstet Invest, 51: 145149. Casanueva E. and Viteri FE. 2003. Iron and oxidative stress in pregnancy. J Nutr, 133: 1700S-1708S. Cederberg J. Siman CM. and Eriksson UJ. (2001). Combined Treatment with Vitamin E and Vitamin C Decreases Oxidative Stress and Improves Fetal Outcome in Experimental Diabetic Pregnancy. Pediatric Research 49: 755-762. Food and Nutrition Board, Institute of Medicine 2000. Dietary reference intake intakes for vitamin C, vitamin E, selenium and caroteniods. Washington DC: National Academy Press, pp.95-185. Halliwell B. and Gutteridge JM. 1990. Role of free radicals and catalytic metal ions in human disease: an overview method in Enzymology (B), 186: 1-85. Hassan GI. and Onu AB. 2006. Total serum vitamin C concentration in pregnant women: implications for a healthy pregnancy Rev Bras Sade Matern Infant, 6: 293296. Kharb S. Gulati N. Ghalaut VS and Singh GP. 2000. Vitamin E concentration in normal pregnant women. J Obstet Gynecol India 50: 48-49.

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J. basic appl. sci. Qanungo S, Mukherjea M. 2000. Ontogenic profile of some antioxidants and lipid peroxidation in human placental and fetal tissues. Mol Cell Biochem, 215: 11-19. Rumbold AC, Crowther CA. 2005. Vitamin C supplementation in pregnancy. The Cochrane Database of Systematic Reviews, 1: CD004072. Walsh SW, Wang Y. 1993a. Deficient glutathione peroxidase activity in preeclampsia is associated with increased placental production of thromboxane and lipid peroxides. American Journal of Obstetrics and Gynecology, 169:1456-1461. Walsh SW, Wang Y. 1998. Placental mitochondria as a source of oxidative stress in pre-eclampsia. Placenta, 19: 581-586. Walsh SW, Wang Y. 1993b. Secretion of lipid peroxides by the human placenta American Journal of Obstetrics and Gynecology, 69: 1462-1466. Wisdom SJ, Wilson R, Mckillop JH, Walker JJ. 1991. Antioxidant systems in normal pregnancy and in pregnancy-induced hypertension American Journal of Obstetrics and Gynecology, 6: 1701-1705. World Medical Association. 2000. World Medical Association Declaration of Helsinki Ethical Principles for Medical Research Involving Human Subjects. 52nd WMA General Assembly, Edinburgh, Scotland. Zachara BA, Wardak C, Didkowski W, Maciag A, Marchaluk E. 1993. Changes in blood selenium and glutathione concentrations and glutathione peroxidase activity in human pregnancy. Gynecol Obstet Invest. 35: 12-17.

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