Abstract
Background and aims: Regarding the increasing numbers of clinical antimonial-resistant Leishmaniasis, understanding the reasons for drug resistance is helpful. This study aimed to find the expression level of the genes related to resistance, P-glycoprotein A (PgpA), G-glutamylcysteine synthetase 1 (Gsh1), and aquaglyceroporin 1 (Aqp1) in antimonial-resistant clinical isolates. Methods: Samples were isolated from leishmaniasis ulcers of 10 non-healing patients and the species were identified by the nested-polymerase chain reaction (PCR) method. In vitro experiments were performed using the amastigote-macrophage model by J774 cell line, and in vivo studies were conducted by animal model, the Balb/c mice. Finally, the values of genes expression were determined by quantitative-reverse transcription (q-RT) real-time PCR method and then compared with non-resistant Leishmania major (MRHO/IR/75/ER). Results: Molecular identification showed that all isolated protozoa were L. major. The isolated samples from clinical resistant patients represented no increase in expression in the tested resistance genes (P>0.05). Finally, it was found that the lack of improvements in patients was not associated with the increased expression of resistance genes. Conclusion: In general, no inherent resistance was observed in the tested samples neither a correlation between the healing of lesions and the level of genes expression.
Highlights
Leishmaniasis is a parasitic disease caused by Leishmania genus of the Trypanosomatidae family in the form of cutaneous (CL), mucocutaneous (MCL), visceral (VL), and post-kala-azar dermal leishmaniasis
The molecular product of the nested-polymerase chain reaction (PCR) on agarose gel 1% showed an amplified band of 560 bp, and in comparison with the standards, L. major (MRHO/IR/75/ ER) (560 bp) and L. tropica (MHOM/IR/02/Mash 10, 750 bp), all samples were suggested to be L. major (Figure 1)
The gene expression of P-glycoprotein A (PgpA), glutamylcysteine synthetase 1 (Gsh1), and aquaglyceroporin 1 (Aqp1) was studied for all isolates
Summary
Leishmaniasis is a parasitic disease caused by Leishmania genus of the Trypanosomatidae family in the form of cutaneous (CL), mucocutaneous (MCL), visceral (VL), and post-kala-azar dermal leishmaniasis. Iran has been identified as an important focus of zoonotic cutaneous leishmaniasis (ZCL) caused by L. major [1,2]. This study aimed to find the expression level of the genes related to resistance, P-glycoprotein A (PgpA), G-glutamylcysteine synthetase 1 (Gsh1), and aquaglyceroporin 1 (Aqp1) in antimonial-resistant clinical isolates. Methods: Samples were isolated from leishmaniasis ulcers of 10 non-healing patients and the species were identified by the nestedpolymerase chain reaction (PCR) method. The values of genes expression were determined by quantitative-reverse transcription (q-RT) real-time PCR method and compared with non-resistant Leishmania major (MRHO/IR/75/ER). The isolated samples from clinical resistant patients represented no increase in expression in the tested resistance genes (P > 0.05).
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