Buthionine sulfoximine increases the efficacy of arteether antimalarial activity in arteether-resistant Plasmodium vinckei by glutathione depletion

Background. L-buthionine (S,R)-sulfoximine (BSO) regulates the glutathione (GSH) level, which in turn exhibits remarkable regulation of several important aspects of cellular metabolism. We hypothesised that increasing the cellular levels of glutathione leads to an increased resistance to arteether, whereas decreasing these by using a GSH inhibitor increases the parasite sensitivity to arteether in the rodent malaria parasite Plasmodium vinckei. Materials and Methods. We tested in vivo effects of BSO on GSH and hemozoin formation in arteether-sensitive and resistant strains. Experimental groups of 7-8 Swiss mice were inoculated by intraperitoneal injection (i.p.) with 1×10 parasitized erythrocytes of PvAS (sensitive) or PvAR (resistant) strain of P. vinckei. The infected mice were treated with BSO (Sigma) 400 mg/kg twice a day for four days and blood was collected after the last injection with BSO. Results. A relatively stronger inhibition of GSH level was observed in the blood of mice infected with resistant parasites (62.64%; p<0.0001), whereas inhibition in sensitive strain-infected mice and uninfected mice was 32% (p=0.034) and 35% (p=0.034), respectively. The results also show an inverse relationship between GSH and hemozoin in the arteether-sensitive and -resistant strains. The hemozoin contents in the resistant strain are 0.27±0.09, 0.69±0.14 and 5.30±0.79 μmol/10 cells at 5, 10 and 20% parasitemia, respectively, whereas hemozoin contents in the sensitive strain at the same parasitemia levels are 0.59±0.29, 12.38±1.96 and 30.80±2.27 μmol/10 cells. Moreover, hemozoin formation increased by 80% through the administration of BSO in the arteether-resistant strain, whereas insignificant changes occurred in the sensitive strain. BSO was also found to increase the efficacy of arteether antimalarial activity against the resistant strain in vivo. Conclusions. Treatment with BSO significantly reduces the level of GSH, which leads to insufficient growth of resistant parasites. These results suggest that BSO might be helpful in prolonging the persistence of the drug, and pose a promising lead to help reducing the chance of resistance development against artemisinin and its derivatives.