Knockdown of Host Antioxidant Defense Genes Enhances the Effect of Glucantime on Intracellular Leishmania braziliensis in Human Macrophages
| Autor: | Ibeth Romero, Maurilio J. Soares, Jair Téllez, Mário Steindel, Alvaro J. Romanha |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Small interfering RNA ATP Binding Cassette Transporter Subfamily B Glutamate-Cysteine Ligase 030106 microbiology Polymerase Chain Reaction Antioxidants Glutathione Synthase Leishmania braziliensis 03 medical and health sciences GSTP1 Meglumine Mechanisms of Resistance Organometallic Compounds Humans Pharmacology (medical) ATP Binding Cassette Transporter Subfamily B Member 1 Pharmacology Gene knockdown Meglumine Antimoniate biology GCLM Macrophages ABCB5 biology.organism_classification Molecular biology Glutathione synthetase Cell biology Infectious Diseases Glutathione Reductase Glutathione S-Transferase pi Host-Pathogen Interactions Thioredoxin |
| Zdroj: | Antimicrobial agents and chemotherapy. 61(7) |
| ISSN: | 1098-6596 |
| Popis: | Leishmaniasis is a neglected tropical disease that affects millions of people worldwide and represents a major public health problem. Information on protein expression patterns and functional roles within the context of Leishmania -infected human monocyte-derived macrophages (MDMs) under drug treatment conditions is essential for understanding the role of these cells in leishmaniasis treatment. We analyzed functional changes in the expression of human MDM genes and proteins during in vitro infection by Leishmania braziliensis and treatment with Glucantime (Sb V ), using quantitative PCR (qPCR) arrays, Western blotting, confocal microscopy, and small interfering RNA (siRNA) human gene inhibition assays. Comparison of the results from gene transcription and protein expression analyses revealed that glutathione S -transferase π1 (GSTP1), glutamate-cysteine ligase modifier subunit (GCLM), glutathione reductase (GSR), glutathione synthetase (GSS), thioredoxin (TRX), and ATP-binding cassette, subfamily B, member 5 (ABCB5), were strongly upregulated at both the mRNA and protein levels in human MDMs that were infected and treated, compared to the control group. Subcellular localization studies showed a primarily phagolysosomal location for the ABCB5 transporter, indicating that this protein may be involved in the transport of Sb V . By inducing a decrease in L. braziliensis intracellular survival in THP-1 macrophages, siRNA silencing of GSTP1 , GSS , and ABCB5 resulted in an increased leishmanicidal effect of Sb V exposure in vitro . Our results suggest that human MDMs infected with L. braziliensis and treated with Sb V express increased levels of genes participating in antioxidant defense, whereas our functional analyses provide evidence for the involvement of human MDMs in drug detoxification. Therefore, we conclude that GSS, GSTP1, and ABCB5 proteins represent potential targets for enhancing the leishmanicidal activity of Glucantime. |
| Databáze: | OpenAIRE |
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