The In Vitro Effect of Hydroalcoholic Extract of Artemisia absinthium on the Growth of Leishmania major (MRHO/IR/75/ER) in Peritoneal Macrophages from BALB/c Mice

Background: Leishmaniasis is a protozoan disease ranging from simple to mucocutaneous, diffuse, and visceral lesions. Current chemical medicines, including Glucantime, have high toxicity, side effects, and drug resistance. Herbal medicines are unlimited sources for discovering new medications to treat infectious diseases. Objectives: This study aimed to examine the in vitro effect of hydroalcoholic extract of Artemisia absinthium on the growth of Leish- mania major in peritoneal macrophages from BALB/c mice. Methods: In this experimental study, the hydroalcoholic extract of A. absinthium was prepared and dissolved in 2% dimethyl sul- foxide (DMSO). Leishmania major (MRHO/IR/75/ER) was cultured in RPMI1640 medium containing 15% fetal bovine serum (FBS). Peri- toneal macrophages were extracted from BALB/c mice and were infected with promastigotes at a ratio of 1:10. The extract was added to Leishmania-infected macrophages and promastigotes at different concentrations. After 24, 48, and 72 hours, the effectiveness of A. absinthium on promastigotes was assessed using Trypan blue test and methyl thiazole tetrazolium (MTT) colorimetric assay, and the half maximal inhibitory concentration (IC50 ) of A. absinthium extract was obtained. Giemsa staining was used to evaluate the effect of A. absinthium extract on amastigotes. Also, inducible nitric oxide synthase (iNOs) level was measured from Leishmania-infected macrophages. The data were analyzed by ANOVA in SPSS software. Results: The hydroalcoholic extract of A. absinthium showed a significant anti-Leishmania activity against promastigotes and amastigotes at various concentrations after 48 and 72 hours. The IC50 values of the alcoholic extract of A. absinthium were obtained at 56 mg/mL and 51 mg/mL concentrations of promastigotes and amastigotes, respectively. The lowest levels of iNOs were seen at 100 and 50 mg/mL concentrations of A. absinthium in treated macrophages. The former was obtained at 60.8 and 58.8 ng/mL after 24 and 48 hours and the latter at 55.5 ng/mL and 50 mg/mL of A. absinthium, respectively, after 72 hours. Conclusions: Findings of the present study showed the anti-parasitic effects of A. absinthium on both promastigotes and amastig- otes. Further studies are recommended to be conducted on animal models and the active ingredients of A. absinthium extract to accurately assess the effects of this extract on the growth of L. major.