Investigation of apoptotic and cytotoxic effects of Hypericin in Leishmania tropica infected macrophage cell line

Objective: In our study, after investigating the in-vitro and ex-vivo antileishmanial effects of hypericin on Leishmania tropica, which is the causative agent of cutaneous leishmaniasis, it was aimed to investigate the effects of active gene regions on the expression levels of active gene regions in the apoptosis mechanism of L.tropica infected THP-1 macrophage cells by qPCR. Materials and Methods: In-vitro, antileishmanial activity (IC50promastigot) of hypericin against L.tropica promastigotes was calculated using CelltTiterglo and hemocytometry methods. The ex-vivo antileishmanial activity of hypericin against L.tropica amastigotes was determined by Giemsa staining method (EC50amastigote). Selectivity index values were calculated with the determined IC50promastigot, EC50amastigote, CC50 values. Expression levels of 88 genes involved in the apoptosis mechanism of THP-1 cells infected with L.tropica were investigated by apoptosis assay, and their effects on gene levels were investigated by qPCR. Results: The CC50 values of hypericin at 24., 48., 72. hours were found to be 40.37-37.69-30.03 µmol, respectively. The in-vitro antileishmanial activity (IC50promastigote) on L.tropica promastigotes was 9.761-9.576-6.994 µmol, respectively, and the ex-vivo antileishmanial activity (EC50amastigote) on L.tropica amastigotes was 25.23-23.12 µmol, respectively. The SI value of hypericin on the two forms of the L.tropica parasite was calculated to be greater than ≥1. After qPCR, genes that were up- and down-regulated in gene expression levels were determined. Conclusions: As a result, it was determined that hypericin showed antileishmanial activity on promastigotes and amastigotes of the L.tropica parasite, and after the apoptosis experiment, hypericin could prevent the escape of L.tropica parasite from the apoptosis mechanism of the host cell. Keywords: Hypericin, Leishmania, Antileishmanial, Cytotoxicity, Apoptosis.