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Evaluation of Apoptotic and Antileishmanial Activities of Artemisinin on Promastigotes and BALB/C Mice Infected with Leishmania major.

Ghaffarifar F, Esavand Heydari F, Dalimi A, Hassan ZM, Delavari M, Mikaeiloo H - Iran J Parasitol (2015 Apr-Jun)

Bottom Line: The in vivo results showed that ointment compounds healed the lesions more effectively rather than intraperitoneal injection method (P<0.05).Interestingly, according to these results it can be concluded that this drug has antileishmanial effects in vitro and in vivo conditions.Artemisinin induces cytotoxic effect on L. major via apoptosis-related mechanism.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.

ABSTRACT

Background: In leishmaniasis, some drugs prescribed for treatment have toxic effects and there are reports about drug resistance in some countries. Due to this fact, using herbal drugs such as artemisinin with good efficacy and low toxic effect might be suitable.

Methods: We evaluated the apoptotic effect of artemisinin on Leishmania major in vitro and the antileishmanial activities of artemisinin on leishmaniasis in BALB/c mice and at the end INF-γ and IL-4 cytokines levels were detected by ELISA in spleen cell culture supernatants. During treatment the lesion size and survival rate were measured each four and ten days, respectively.

Results: Percentage of early and late apoptosis in promastigotes of control group and promastigotes treated with 10, 25, 50 and 100 μg/ml of artemisinin after 48 h were 0.13, 16.04, 41.23, 49.03 and 81.83, respectively. The IFN-γ in ointment treated group were higher than those of other groups (P<0.05). The in vivo results showed that ointment compounds healed the lesions more effectively rather than intraperitoneal injection method (P<0.05). The survival rate of mice 150 days after challenge in treated group with ointment of artemisinin was 66% while all mice in control groups were died.

Conclusion: All of in vitro results represented that this drug had antileishmanial effects and these results were confirmed by evaluation effects in vivo condition of leishmaniasis. Interestingly, according to these results it can be concluded that this drug has antileishmanial effects in vitro and in vivo conditions. Artemisinin induces cytotoxic effect on L. major via apoptosis-related mechanism.

No MeSH data available.


Related in: MedlinePlus

Flow cytometry results of the effect of artemisinin with 10, 25, 50, and 100 μg/ml concentrations on the promastigotes (2×106 promastigotes /ml) of Leishmania major (MRHO/IR/75/ER) viability and comparing them with control group (promastigotes of Leishmania major without any treatment) and macrophages treated with 25 μg/ml of artemisinin (IC50 dose on promastigotes) after 48 h. Regions of quadrate show percentage necrosis, late apoptosis, apoptosis and live cells
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Figure 3: Flow cytometry results of the effect of artemisinin with 10, 25, 50, and 100 μg/ml concentrations on the promastigotes (2×106 promastigotes /ml) of Leishmania major (MRHO/IR/75/ER) viability and comparing them with control group (promastigotes of Leishmania major without any treatment) and macrophages treated with 25 μg/ml of artemisinin (IC50 dose on promastigotes) after 48 h. Regions of quadrate show percentage necrosis, late apoptosis, apoptosis and live cells

Mentions: Induction of apoptosis in promastigotes of L. major was analyzed by flow cytometry FACS Calibor system after staining with Annexin-V and PI. The percentages of apoptotic, necrotic and viable cells were determined: UL as necrotic cells in the upper left region (positive for PI), UR as the cells were banded with Annexin-V and PI in upper right region, LL as viable promastigotes in lower left region and LR as a marker of apoptosis in the lower right region of figures. The results were shown that artemisinin were induced apoptosis in promastigotes of L. major at all concentrations of artemisinin. The most effects of artemisinin on promastigotes of L. major took place in apoptotic areas. The percentages of apoptosis in promastigotes stimulated with 10, 25, 50 and 100 μg/ml of artemisinin were calculated 12, 25.90, 40.52 and 68.16 respectively in comparison with control group that was 0.02. The results of necrosis, late apoptosis, apoptosis and live promastigotes are shown in Fig. 3.


Evaluation of Apoptotic and Antileishmanial Activities of Artemisinin on Promastigotes and BALB/C Mice Infected with Leishmania major.

Ghaffarifar F, Esavand Heydari F, Dalimi A, Hassan ZM, Delavari M, Mikaeiloo H - Iran J Parasitol (2015 Apr-Jun)

Flow cytometry results of the effect of artemisinin with 10, 25, 50, and 100 μg/ml concentrations on the promastigotes (2×106 promastigotes /ml) of Leishmania major (MRHO/IR/75/ER) viability and comparing them with control group (promastigotes of Leishmania major without any treatment) and macrophages treated with 25 μg/ml of artemisinin (IC50 dose on promastigotes) after 48 h. Regions of quadrate show percentage necrosis, late apoptosis, apoptosis and live cells
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4522302&req=5

Figure 3: Flow cytometry results of the effect of artemisinin with 10, 25, 50, and 100 μg/ml concentrations on the promastigotes (2×106 promastigotes /ml) of Leishmania major (MRHO/IR/75/ER) viability and comparing them with control group (promastigotes of Leishmania major without any treatment) and macrophages treated with 25 μg/ml of artemisinin (IC50 dose on promastigotes) after 48 h. Regions of quadrate show percentage necrosis, late apoptosis, apoptosis and live cells
Mentions: Induction of apoptosis in promastigotes of L. major was analyzed by flow cytometry FACS Calibor system after staining with Annexin-V and PI. The percentages of apoptotic, necrotic and viable cells were determined: UL as necrotic cells in the upper left region (positive for PI), UR as the cells were banded with Annexin-V and PI in upper right region, LL as viable promastigotes in lower left region and LR as a marker of apoptosis in the lower right region of figures. The results were shown that artemisinin were induced apoptosis in promastigotes of L. major at all concentrations of artemisinin. The most effects of artemisinin on promastigotes of L. major took place in apoptotic areas. The percentages of apoptosis in promastigotes stimulated with 10, 25, 50 and 100 μg/ml of artemisinin were calculated 12, 25.90, 40.52 and 68.16 respectively in comparison with control group that was 0.02. The results of necrosis, late apoptosis, apoptosis and live promastigotes are shown in Fig. 3.

Bottom Line: The in vivo results showed that ointment compounds healed the lesions more effectively rather than intraperitoneal injection method (P<0.05).Interestingly, according to these results it can be concluded that this drug has antileishmanial effects in vitro and in vivo conditions.Artemisinin induces cytotoxic effect on L. major via apoptosis-related mechanism.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.

ABSTRACT

Background: In leishmaniasis, some drugs prescribed for treatment have toxic effects and there are reports about drug resistance in some countries. Due to this fact, using herbal drugs such as artemisinin with good efficacy and low toxic effect might be suitable.

Methods: We evaluated the apoptotic effect of artemisinin on Leishmania major in vitro and the antileishmanial activities of artemisinin on leishmaniasis in BALB/c mice and at the end INF-γ and IL-4 cytokines levels were detected by ELISA in spleen cell culture supernatants. During treatment the lesion size and survival rate were measured each four and ten days, respectively.

Results: Percentage of early and late apoptosis in promastigotes of control group and promastigotes treated with 10, 25, 50 and 100 μg/ml of artemisinin after 48 h were 0.13, 16.04, 41.23, 49.03 and 81.83, respectively. The IFN-γ in ointment treated group were higher than those of other groups (P<0.05). The in vivo results showed that ointment compounds healed the lesions more effectively rather than intraperitoneal injection method (P<0.05). The survival rate of mice 150 days after challenge in treated group with ointment of artemisinin was 66% while all mice in control groups were died.

Conclusion: All of in vitro results represented that this drug had antileishmanial effects and these results were confirmed by evaluation effects in vivo condition of leishmaniasis. Interestingly, according to these results it can be concluded that this drug has antileishmanial effects in vitro and in vivo conditions. Artemisinin induces cytotoxic effect on L. major via apoptosis-related mechanism.

No MeSH data available.


Related in: MedlinePlus