Effect of cantharidin on apoptosis of the Leishmania major and on parasite load in BALB/c mice.

摘要

Leishmania major is causative agent of cutaneous leishmaniasis. Leishmaniasis is most public health problem in developing countries. Cantharidin is vesicant found in the Meloidae beetles hemolymph. Protein phosphatase 1 and 2A is the primary target of cantharidin. Cantharidin induce apoptosis in variant cancer cells. In this study, the effect of various concentrations of cantharidin on the Leishmania major viability was investigated in vitro by flow cytometry and MTT assay. Cantharidin also was applied as ointment by three doses (0.05, 0.5 and 0.1%) on leishmaniasis lesion in BALB/c for 4 weeks, quantity of parasite assessed by Real-Time PCR. MTT test results indicated that 0.5 micro g mL-1 and 50 micro g mL-1 cantharidin had 14.26 and 49.86% cytotoxicity in leishmania promastigotes after 72 h, respectively. But, flow cytometry data showed 14.29% cytotoxicity (as 13.12% apoptosis, 1.12% late apoptosis and 0.05% necrosis) and 68.51% cytotoxicity (as 5.28% apoptosis, 63.23% late apoptosis and 0% necrosis) in promastigotes due to 0.5 and 50 micro g mL-1 cantharidin after 72 h, respectively. Mean of parasite load in control, 0.05, 0.1 and 0.5% cantharidin-treated groups were 85000, 2400, 2200 and 250 parasite/ micro L, respectively. It can be concluded that cantharidin induces apoptosis in the Leishmania major promastigotes and infected macrophages in the time and dose-dependent manner; also it can decrease parasite load in infected mice.