Evaluation of curcumin and CM11 peptide alone and in combination against amastigote form of Iranian strain of L. major (MRHO/IR75/ER) in vitro

Abstract

Curcumin (diferuloylmethane) is the main phytochemical of Curcuma longa Linn, an extract of the rhizome turmeric. For thousands of years, turmeric among other natural products has been used as a dietary spice and as a medicinal plant in Asian countries. The present study reports the leishmanicidal activity of curcumin in different concentrations (10 μM, 20 μM, 40 μM). It is also showing the effect of CM11 peptide (8 μM) alone and in combination with curcumin (10 and 20 μM) as a leishmanicidal drug. The experiments were performed with the amastigote form of Leishmania major (MRHO/IR/75/ER) in vitro and the leishmanicidal activity was analyzed after 12 and 24 h of incubation by Giemsa and DAPI staining. Further investigation was done by using semi-quantitative PCR with new designed common primer pair derived from an 18S rRNA gene belonging to the L. major and mouse, which amplified the above-mentioned gene segments simultaneously with different PCR product size. Our findings showed that curcumin had leishmanicidal activity in a dose and time-dependent manner and its lowest effective dose was at concentrations of 40 μM afetr12 h and 10 μM after 24 h. The IC50 value of curcumin against amastigote forms of L. major was 21.12 μM and 11.77 μM after 12 and 24 h, respectively. Treatment of amastigote form with CM11 (8 μM) alone and in combination with curcumin (10 μM and 20 μM) showed less leishmanicidal activity. Interestingly, CM11 in combination with curcumin (10 μM and 20 μM) had even less leishmanicidal effect compared to curcumin alone in the same concentrations (10 μM and 20 μM). The semi-quantitative PCR analysis confirmed the data achieved by Giemsa and DAPI staining and showed that curcumin reduced the PCR product derived from amastigote form in concentration and time-dependent manner compared to the genome of the host cells.