RESUMEN
Leishmaniasis is a major health problem worldwide with different clinical forms that depend on the parasite, the host's immune system, and immune-inflammatory responses. This study aimed to evaluate the secondary metabolites from Artemisia kermanensis Podlech by bioguided fractionation against Leishmania major. The chemical structures of the isolated compounds were determined based on analysis of mass and nuclear magnetic resonance spectra. Antileishmanial activity were determined on promastigotes and amastigotes. Chemical structures of the isolated compound were as 1-Acetoxy-3,7-dimethyl-7-hydroxy-octa-2E,5E-dien-4-one for compound 1 and 5,7-dihydroxy-3',4',6-trimethoxyflavone (Eupatilin) for compound 2, and 5,7,3'-Trihydroxy-6,4',5'-trimethoxyflavone for compound 3. Compound 2 were confirmed by significant activity with IC50 of less than 50 µg/ml for 24 and 48 h in clinical form (amastigotes). Compound 3 demonstrated high susceptibility with an IC50 of less than 30 µg/ml for promastigotes for 24 h. The bioguided fractionation of A. kermanensis resulted the isolation of potent antileishmanial agents with a low toxicity effect on macrophages. These plant metabolites can be a candidate as a drug for treating cutaneous leishmaniasis.
RESUMEN
BACKGROUND: Leishmaniasis is a skin disease caused by Leishmania parasite. Despite being self-limiting, must be treated. Available drugs have side effects and drug resistance has also been seen. MATERIALS AND METHODS: Maggot debridement therapy (MDT) is using sterile fly larvae (maggots) of blow flies (Lucilia sericata) for the treatment of different types of tissue wounds. Larvae have excreted and secreted substances that have been proved to have antimicrobial effects, in addition to the some other specifications. RESULTS: In this study, the anti-leishmanial effects of extracts and secretions of sterile second- and third-instar larvae of L. sericata on the growth of Leishmania major promastigotes and amastigotes in the J774 macrophages have been evaluated in vitro. CONCLUSION: The results showed that extracts and secretions had almost the same leishmaniocidal effect on promastigotes and intracellular amastigotes without cytotoxic effect on macrophages.
RESUMEN
BACKGROUND: Resistance of Leishmania species to antimonial drugs has increased. Hence, in the present study Leishmania major isolates were collected from patients with resistance phenotype and the presence/absence of resistance to Glucantime was investigated. MATERIALS AND METHODS: Samples were taken from 10 cutaneous leishmaniasis (CL) patients who had not responded to chemotherapy with Glucantime. Nested polymerase chain reaction (PCR) was performed to identify the isolated species. Stationary phase promastigotes were added to the grown, adhesive J774 macrophages. Values obtained from standard strain were compared with the test cultures after exposure to the medicine. In vivo, the effects of Glucantime were assessed by comparing the sizes and the parasite burden of the lesions on mouse model. RESULTS: The results of amplified band on agarose gel demonstrated all samples were L. major. After exposure to medicine, a reduction of intracellular amastigotes to half was detected. In vivo, the parasite was eliminated in 90% of mice with lesions caused by both isolates of patients and standard L. major, and their lesions became smaller significantly. CONCLUSION: Pentavalent antimonial (SbV) salts are the main component of chemotherapy against leishmaniasis. However, the medicine has been found ineffective. In the present study, isolates from patients with no response to treatment had no significant difference from the standard L. major strain (as the sensitive strain). Therefore, in patients with resistance phenotype to Glucantime, the parasites did not actually have intrinsic resistance, i.e., environmental and host factors prevented the successful treatment of the disease.
RESUMEN
BACKGROUND: Leishmaniasis is a major health problem in some endemic areas of tropical and subtropical areas of the world. Interleukin-12 (IL-12) and interferon gamma (IFN-γ) are essential cytokines associated with initiation of Th1 response. The main objective of this study was to evaluate of the type of immune response to L. major isolates from patients with no clinical response to antimonite (Glucantime). MATERIALS AND METHODS: This experimental study was carried out during 2013-2014. In the current study Leishmania major were isolated from 10 CL patients with a history of at least one course of treatment with Meglumine antimonate (Sb5). The isolates were used to evaluate in vitro and in vivo response to Sb5. J774 murine macrophage cell line was used for in vitro tests and Balb/c mice was used for in vivo studies. IL-12 gene expression was evaluated using Real-time PCR and IFN-γ serum level was quantified using ELISA technique. SPSS (version: 20), analysis of Covariance (ANCOVA) was used for statistical analysis. RESULTS: PCR results confirmed that all 10 isolates were L. major. The mean of IL-12 gene expression in vitro, in vivo and IFN-γ serum levels (pg/ml) after 2 and 3 weeks treatment in vivo, increased significantly following the treatment with Glucantime in the two groups of Balb/c mice infected either with patients' isolates or standard L. major. No significant difference was seen between the patients' isolates and standard species. CONCLUSIONS: Although the L. major were isolated from patients with active lesion and no clinical response to Glucantime after at least one courses of Glucantime treatment but in vivo and in vitro immune response of L. major isolates showed no difference between the patients' isolates and standard L. major.
RESUMEN
BACKGROUND: Leishmaniasis is a parasitic disease caused by different species of Leishmania parasites with a wide range of clinical manifestations. Antimonial compounds such as meglumine antimoniate (glucantime) are the first line drugs for the treatment of leishmaniasis. However, according to reports of the drug resistance of parasites, the efficacy of antimonial compounds is low. The ATP-binding cassette (ABC) proteins are present in all organisms and mediate the transport of vital elements through biological membranes. One of the important mechanisms of resistance in Leishmania parasites is the overexpression of ABC efflux pumps. P-glycoprotein A (pgpA) is a related gene for ABC transporter in Leishmania species. The aim of this study was to compare the pgpA expression in laboratory-induced resistant L. major (MRHO/IR/75/ER) and sensitive parasites. METHODS: RNA extraction of promastigotes of sensitive and resistant clones was performed and total RNA was reverse transcribed. The real-time quantitative polymerase chain reaction (PCR) was used to assess RNA expression profiles and the expression levels were calculated using 2(-ΔCt) method. RESULTS: The mean expression level of pgpA mRNA was 2.70 ± 0.51 in in sensitive Leishmania clone and 6.08 ± 1.50 in resistant Leishmania clone (P = 0.021). CONCLUSION: The expression of pgpA gene in resistant strains of L. major was almost fivefold higher than those in susceptible strains. Therefore, this can be used in field isolates, i.e. overexpression of the gene can prove resistance in wild type field isolates.