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Environmentalmeasures for enhancing public health: chloroquine

Biology and Medicine, 1 (3): 39-43, 2009 eISSN: 09748369,
The influence of chloroquine administration on antioxidant levels,
oxidant marker and total cholesterol in Wistar rats
AC Achudume
Institute of Ecology and Environmental Studies, Obafemi Awolowo University, Ile-Ife, Nigeria.
This study was undertaken to determine some biochemical changes of importance associated with chloroquine (CQ)
treatment, using rat model of Plasmodium beighei berghei parasites. Chloroquine phosphate (5mg/kg body wt,
dissolved in distilled water) was administered 3days per week for 8 weeks after infection by Plasmodium beighei
beighei. The chloroquine did not potentiate any adverse effect on the formation of ascorbic acid-linked lipid
peroxidation. The drug increased glutathione contents, but decreased protein and cholesterol levels. Chloroquine
alone had no significant effect on malondialdehyde and alanine/aspartate transaminases, but decreased protein
synthesis and cholesterol level. The characteristic biologic effects of CQ including the decrease in total cholesterol,
protein synthesis, and stabilizer of lipid peroxidation as measured by malondialdehyde and the increased level of
glutathione, may enhance the environmental measures of public health in cholesterol compromised individuals.
Keywords: Chloroquine, cholesterol, hypercholesterolemia, roll-back malaria, public health, ascorbic acid-linked lipid
peroxidation, hepatic transaminases.
the assessment of risks entailed by regular use Chloroquine (CQ) has become increasingly of chloroquine and (b) lack of resources for ineffective largely because of development of assessment of chloroquine for which data are at resistance to the drug by malaria parasites. Massive use of this drug to help eradicate resistance to malaria infection has a multigenic selective pressure, drug-resistance strain of mechanism as widely believed (Schwartz 2003), Plasmodium falciparum malaria emerged that this association may govern other mechanism gradually flourished (Wellems et, al., 2001, that has potential benefit in cholesterol reduction Medilinks 2002, and WHO/UNICEF 2003). The Achudume and Eno 2000). Bioaccumulation and chloroquine and some other older antimalarial toxicity of chloroquine in lysosomal cells and drugs prompted the introduction of more based liver are well documented (Korolenko et. al., effective newer drugs such as the artemisinin- 1990). Our previous investigations showed that combination therapy as well as the concept of chloroquine deaccelerated NADPH-linked lipid roll-back malaria (Li et. al., 2003 and Yamey peroxidation (Achudume, 1997). Similar reports showed that CQ causes alterations in lipid metabolism (Achudume et. al., 1998) and essential to meet the social and economic goals inhibition of cholesterol synthesis in freshly of the endemic community and today’s best isolated rat hepatocyte (Sewell et. al., 1983). practice demonstrates that it can be used widely Taking together, these results provide strong in a cost-effective manner and with a high evidence that CQ may still be valid as a drug for degree of safety. However, a great deal remains to be done to ensure environmental health and glutathione (GSH) and hepatic enzymes [alanine sustainable development and improved quality (ALT) and aspartate (ASP) transaminases] as indices of toxicity, exposure to low-level CQ problems, particularly in developing countries, was investigated as to whether it has potential to are (a) lack of sufficient scientific information for Biology and Medicine, 1 (3): 39-43, 2009 cholesterol compromised individuals. Many suppression of parasitaemia was calculated for workers had attempted dietary approaches each day by comparing the parasitaemia in (DASH) to lower cholesterol level which may infected control with those of treated rats reverse hypertension (Obarzanek et. al., 2001, (Ayaiyeola et. al., 2006). The positive control Ignatus et al 2006). Other study used Satin for received infected erythrocytes and negative the treatment of cholesterol (Caspard 2006) and all these treatments have their drawbacks. They either have long-term cardiac risk or elevate liver decapitation. Livers were dissected out and enzymes and cause myopathy as well as other homogenized in TRI-R-STIR-R model K-43, and side effects. None had studied the prophylactic effects of CQ on lipid peroxidation, cholesterol, GSH and liver enzymes to justify calls for public health strategies to maximize the useful life of The supernatant was dialyzed against ice cold CQ. In the present study, an assessment of distilled water and resuspended in 0.15 M KCl to chloroquine as alternative to malaria treatment an approximately final concentration of 20 mg of were determined by measuring the levels of protein per millimeter (approximately 1g of liver antioxidant (GSH) and oxidant marker (MDA), per millimeter (Siegler and Kararinoff 1983). The and total cholesterol levels by analyzing percent parasitemia to the liver enzymes in infected thiobarbiturate. The reaction was initiated by addition of ascorbic acid (Shimada et. al., 1979). Materials and Methods
Reduced glutathione was determined by using aliquots of liver homogenate (20%) diluted with Adult male albino Wistar rats weighing between 190-200g were obtained from animals holding, nitrobenzoic acid) was added to the supernatant University, Ile-Ife. Animals were housed in fractions and absorbance was measured at ventilated group aluminum boxes (41x28x15 cm) and illumination (12h light cycle starting at 6AM) modified by Liu et. al. (2005). Total cholesterol for at least 7 days before experiments. Animals level was determined by using Liebermann were maintained on Laboratory chow (Ladokun Burechard reagent (Abell et. al., 1962). Protein was determined by Lowry et. al. (1951) method. deprived of food for 24h but given free access to water up to the beginning of the study. Protocols describing the use of rats were approved by the An ANOVA fol owed by Dunnett’s test was used Animal Care Committee of OAU, Ile-Ife and in to compare treated groups to a control group, accordance with the American Physiological after verification of homoscedasticity and equal Society’s “Guiding Principles for Research Results and Discussion
The effects of CQ administration on ascorbic All chemicals used were of analytical grade. acid-linked lipid peroxidation, protein contents, glutathione, total cholesterol, alanine and dissolved in distilled water) was administered aspartate transaminases is summarized in Table orally for 3days before and after infection, i.e. 3 1. Chloroquine treatment for eight weeks did not days per week for 8 weeks. The experimental potentiate any adverse effect on the formation of animals received a standard inoculum of 1x107 malondialdehyde in ascorbic acid-linked lipid peroxidation. Chloroquine alone decreased erythrocytes intraperitoneally (ip), following the protein synthesis significantly P<0.05 and methods of Makinde et. al. (1993). Blood increased glutathione content. It has little or no schizontocidal activity (Rane test) was evaluated effect on the hepatic enzymes and significantly after 3, 5, and 8 days post infection. This was (P<0.05) decreased cholesterol level. In a monitored by collection of drops of blood from similar manner, Chloroquine with infected blood the tail vein, followed by fixation, permeability, staining and analyzed with a visible range flow compared to control P<0.05. It increased cytometer (Barkan et. al., 2000). The percentage Research Article Biology and Medicine, Vol 1 (3): 00-00, 2009 Table 1: Effect of chloroquine administration on ascorbic acid-linked lipid peroxidation in subcellular fraction of rat liver Treatment Malondialdehyde Protein Glutathione Cholesterol ALT ASP 8 weeks nmol/30 min mg mg/ml mg/ml mg/100 ml mg/protein protein Control 0.12±0.1 13.29±0.62 0.300±0.10 12.65±2.49 0.33±0.020 .30±0.01 CQ plus Infected Blood 0.382±0.5 1.522±0.50x 0.843±0.21 2.51±8.1x 0.29±0.06 0.29±0.09 CQ alone 0.494±0.41 1.783±0.41x 0.735±0.26 2.32±1.6x 0.36±0.08 0.34±0.06 x Significantly different from control P<0.05 Table 2: Schizontocidal activity of Chloroquine on early infection Treatment Dose Average % chemosuppression Mg/kg/day 3 5 8 days Control - 0 0 0 CQ 5 25±1.2 58.6±0.6 86.6±1.3 hepatic enzymes and significantly decreased cholesterol level. The schizontocidal action of chemosuppressive activity of Chloroquine was CQ assessed its chemosuppressive activity is greatly diminished (Table2). The percentage chemosuppression was observed to increase by described in literature (Makinde et. al., 1993). synthesis, reduced cholesterol level while Although the results presented did not relate increasing reduced glutathione concentrations. It sensitivity of CQ to resistant factors. The study had no significant effect on ascorbic acid-linked however, showed that CQ treatment did not lipid peroxidation and ALT/AST transaminases show any adverse effect on the formation of Biology and Medicine, 1 (3): 39-43, 2009 malondialdehyde in ascorbic acid-linked lipid Achudume, A.C., Nwoha, P.U., Olarenwaju, I.G. peroxidation. The decrease in cholesterol levels 1997. Influence of a low protein diet on serum may be the result of CQ- accelerated excretion of cholesterol through feces, after chloroquine peroxidation in Chloroquine treated rats. International Journal of Food Science and Nutrition, 48; 339-343 interruption of the entero-hepatic recycling of Ajaiyeola, E., Falade, M., Ogbole, O., Okpako, L., Akinboye, D. 2006. In vivo antimalarial and cytotoxic administration resulted in a modest impairment properties of Annona senegalensis extract. African of cholesterol reabsorption and in a possible Journal of Traditional, Complementary and Alternative alteration in binding and permeability properties of various membrane systems resulting in excretion of proteins. Several studies have Barkan, D., Ginsburg, H., Golenser, J., 2000. shown that CQ binds quantitatively to membrane Optimization of low cytometric measurement of parasitaemia system and reduces the binding capacity of the International Journal of Parasitology, 30(5); 649-653 plasma membrane through derangement of the recycling of receptors (Wellems et. al., Beynen, A.C. 1986. Could Chloroquine be of value in 1991).This result is an extension of the the treatment of hypercholesterolemia? Artery 13; observation of Sewell (1983) and Achudume (1997), that cells exposed to CQ take up and concentrate this drug, resulting in an extensive Caspard, H., Chan A.K., Walker, A.M. 2006. Cholesterol after initiation of Satin Treatment. The Dushkin, M.I., Dolgov, A.V. 1986. Activity of key synthesis may not entirely due to the products of enzymes of cholesterol biotransformation and lipid lipid peroxidation, since, CQ is effectively contents in the liver, aorta, adrenals and blood of rats scavenged by glutathione. GSH deceased the stimulatory action of ascorbic acid-linked lipid prevents protein synthesis. Accordingly, the Foote, S.J., Cowman, A.F. 1994. The mode of action and the mechanism of resistance to antimalarial reduction of cholesterol level as a result of CQ ingestion and the high levels of glutathione, CQ Ignatus, G.E., Zarraga, M.D., Ernst, R. Schwarz, M.D. may find some uses in individuals with history of 2006. Impact of dietary patterns and interventions in excessive accumulations of cholesterol. Though cardiovascular health. Circulation 114: 961-973 chloroquine may be a resistant drug in malaria infection, it is nevertheless useful in cholesterol Korolenko, T.A, Rukavishnikova, E.V., Pupyshev, A.B repeated administration of Chloroquine on the activity of lysosomal proteinases in rat liver cells. Vopr. Med. References
Abell, L.L, Levy, B.B., Brodie, B.B., Kendall, F.E. Li, Q.G., Si, Y.Z, Lee, P., Wong, E., Xie, L.H., Kyle, 1962. Simplified method for the estimation of D.E., Dow, G.S. 2003. Efficacy comparison of cholesterol in serum and demonstration of its intravenous artelinate and artesunate in Plasmodium specificity. Journal of Biological Chemistry, 196; 366- berghei. Parasitology, 126; 283-291 Liu, G.T., Li, Y., Wei, H.L., Zhang, H., Xu, J.Y., Yu, L.H. 2005. Mechanism of protective action of bicyclol Olarenwaju, I.G. 1998. Experimental analysis of against CCl4-induced liver injury in mice. Liver gossypol and chloroquine interaction in serum and liver of rat. Indian Journal of Experimental Biology, 36; Lowry, G.B., Rosebrough, N.J., Fair, A.L, Randoll, A.J. 1951. Protein measurement with Folin Phenol Achudume, A.C., Eno, T.L. 2000. Distribution of key reagent. Journal of Biological Chemistry, 93; 265-275 ions of chloroquine biotransformation and cholesterol uptake in rat liver. International Journal of Food Makinde, J.M., Awe, S.O., Salako, L.A. 1993. Seasonal variation in the antimalarial activity of Biology and Medicine, 1 (3): 39-43, 2009 Morinda lucida on Plasmodium berghei berghei in immune traveler to East Africa. Clinical Infectious Shimada, O. Yasuda, H. 1979. Lipid peroxidation and Chloroquine is energy-dependent. Health News its inhibition by teronidine 11. Ascorbic acid induced Biochemistry Biophysics Acta, 572; 531-536 Obarzanek, E., Sacks, F.M., Vollmer, W.M., Bray, G.A, Miller, E.R, Lin, P.H, Karanja, N.M, Most- Siegler, J.H., Kazarinoff, M.N. 1983. The effect of Windhauser, M.M., Moore, T.J., Swain, J.F, Bales, acute and chronic protein deprivation on ornithine C.W, Proscham, M.A, 2001. Effects on blood lipids of decarboxylase 1 in rat liver and colon. Journal of Approaches to Stop Hypertension (DASH). Nutrition Wellems, T.E., Plowe, C.V. 2001. Chloroquine resistance malaria. International Infectious Disease implications. Annals of European Pharmacology and Wellems, T.E., Walker-Jonah, A.S., Panton, L.S 1991. Understanding the biochemical properties and Sewell, R.B., Barham, S.S., LaRusso, N.F. 1983. functional role of CQ. Proceedings of the National Effect of Chloroquine on the form and function of hepatocyte lysosomes. Morphologic modifications and physiologic alterations related to the biliary excretion WHO/UNICEF, 2003. The Africa Malaria Report of lipids and proteins. Gastroenterology, 85; 1146- Yamey, G. 2004. Roll-back malaria: a failing global Schwartz, E., Bujanover, S., Kain, K.C. 2003. Genetic health campaign. British Medical Journal, 328; 1086- Plasmodium falciparum malaria acquired by a non-


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