THE PROTECTIVE EFFECTS OF KOLAVIRON ON ARTEMISININ COMBINATION ANTIMALARIA-INDUCED TOXICITY IN THE RAT

Abstract

Artemether-Lumefantrine (AL), Artesunate-Mefloquine (AM) and Artesunate-Amodiaquine (AA) are widely used Artemisinin Combination Antimalarial (ACA). These drugs induce toxicity in experimental animals through generation of free radicals and hence there is the need to ameliorate this toxicity. Kolaviron is a biflavonoid from Garcinia kola known to offer protection against oxidative damage in vivo. The study was designed to investigate the effects of Kolaviron on ACA-induced toxicity in the rat. Two hundred and forty male Wistar rats (180-220g) were divided equally into eight groups, viz: control, AL-treated (19.2mg/kg), AM-treated (Artesunate-11.4mg/kg and Mefloquine-10.8mg/kg), AA-treated (Artesunate-11.8mg/kg and Amodiaquine-20mg/kg), Kolaviron (100mg/kg), Kolaviron + AL, Kolaviron + AM and Kolaviron + AA-treated animals. Blood samples (4-5mL) were collected into heparinized tubes and plasma obtained was used to determine the levels of urea, creatinine, bilirubin, Alanine amino Transferase (ALT), Aspartate amino Transferase (AST), Total-Cholesterol (TC), High Density Lipoprotein-Cholesterol (HDL-C) and Triglycerides by spectrophotometry.The Low Density Lipoprotein-Cholesterol (LDL-C) was obtained by Friedewald formula. Post-mitochondrial fraction of liver was used to evaluate the levels of Lipid Peroxidation (LPO), Superoxide Dismutase (SOD), Catalase, Glutathione-S-Transferase (GST), Glutathione Peroxidase (GPx), Aniline Hydroxylase (AH), Vitamin C (VC) and Glutathione (GSH) by spectrophotometry. Genotoxicity was evaluated by measuring Micronucleated Polychromatic Erythrocytes (mNPCEs) in bone marrow cells. Data were analyzed using ANOVA and Students’t-test at p=0.05. Hepatic GST (9.2 ± 0.2, 11.2 ± 0.3 and 11.6 ± 0.3nmols/min/mg protein), GPx (0.80 ± 0.02, 0.90 ± 0.03 and 1.10 ± 0.03 nmols/min/mg protein) and AH (0.13 ± 0.01, 0.15 ± 0.01 and 0.16 ± 0.02μmols/mg protein) activities in AL, AM and AA- treated rats were reduced when compared to the control (16.2 ± 0.5nmol/min/mg protein, 1.6 ± 0.03nmol/min/mg protein, and 0.2 ± 0.02μmols/mg protein) respectively. Rats treated with AL, AM and AA had higher values of urea (54.0 ± 3.0, 43.0 ± 1.0 and 41.0 ± 2.0 mg/dL), creatinine (0.60 ± 0.02, 0.48 ± 0.01 and 0.40 ± 0.04mg/dL), bilirubin (0.72 ± 0.02, 0.60 ± 0.01 and 0.52 ± 0.02mg/dL), ALT (81.0 ± 1.0, 66.0 ± 2.0 and 57.0 ± 3.0 U/L) and AST (88.0 ± 1.0, 68.0 ± 2.0 and 55.0 ± 3.0 U/L) relative to the control (32.0 ± 2.0, 0.32 ± 0.01, 0.38 ± 0.01, 48.0 ± 1.0 and 42.0 ± 2.0) respectively. Pre-treatment of AL, AM and AA- treated rats with kolaviron caused reduction in the levels of urea, creatinine, bilirubin, ALT and AST relative to vi untreated rats. The increase in lipid profile and LPO in rats treated with AL, AM and AA (TC- 62.0 ± 2.0, 52.0 ± 1.0 and 49.0 ± 2.0 mg/dL, HDL-C- 26.0 ± 2.0, 21.0 ± 2.0 and 18.0 ± 1.0mg/dL, LDL-C- 42.0 ± 1.0, 37.0 ± 1.0 and 34.0 ± 2.0mg/dL, Triglycerides- 56.0 ± 3.0, 50.0 ± 2.0 and 41.0 ± 1.0mg/dL and LPO- 650.0 ± 4.0, 509.0 ± 2.0 and 488.0 ± 3.0 nmol/mg protein) was reduced by kolaviron pretreatment (TC- 48.0 ± 2.0, 42.0 ± 1.0 and 40.0 ± 2.0 mg/dL, HDL-C- 19.0 ± 1.0, 16.0 ± 1.0 and 13.0 ± 1.0mg/dL, LDL-C- 35.0 ± 2.0, 32.0 ± 1.0 and 28.0 ± 1.0mg/dL, Triglycerides-47.0 ± 2.0, 44.0 ± 2.0 and 39.0 ± 1.0mg/dL and LPO- 432.0 ± 3.0, 351.0 ± 2.0 and 328.0 ± 5.0 nmol/mg protein). Pre-treatment with kolaviron increased the activities of SOD, catalase, levels of VC and GSH when compared with untreated rats. Administration of kolaviron reduced the frequency of mNPCEs (15.0 ± 0.2, 13.0 ± 0.3 and 11.0 ± 0.2/1000PCEs) when compared with untreated rats (21.0 ± 0.4, 16.0 ± 0.2 and 15.0 ± 0.3 mNPCEs/1000PCEs). Kolaviron protected against Artemether-Lumefantrine, Artesunate-Mefloquine and Artesunate-Amodiaquine- induced damages in the liver and kidney of the rat. Antioxidant properties of Kolaviron may be responsible for these protective effects.