P-glycoprotein A Gene Expression in Glucantime-Resistant and Sensitive Leishmania major (MRHO/IR/75/ER)
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.
Alvar J, Vélez ID, Bern C, Herrero M, Desjeux P, et al. Leishmaniasis Worldwide and Global
Estimates of Its Incidence. PLoS ONE. 2012; 7(5):e35671.
Firooz A, Khamesipour A, Ghoorchi MH, Nassiri-Kashani M, Eskandari SE, Khatami AR, Hooshmand B, Gorouhi F, Rashighi- Firoozabadi M, Dowlati Y. Imiquimod in combination with meglumine antimoniate for cutaneous leishmaniasis: A randomized assessor-blind controlled trial. Arch Dermatol. 2006; 142:1575-1579.
Shazad B, Abbaszadeh B, Khamesipour A. Comparison of topical paromomycin sulfate (twice/day) with intralesional meglumine antimoniate for the treatment of cutaneous leishmaniasis caused by L. major. European J Dermatol. 2005; 15(2):85-7.
Hadighi R, Mohebali M, Boucher P, Hajjaran H, Khamesipour A, Ouellette M. Unresponsiveness to Glucantime treatment in Iranian cutaneous leishmaniasis due to drug-resistant Leishmania tropica parasites. PLoS Med. 2006; 3:162.
Légaré D, Cayer S, Singh A K, Richard D, Papadopoulou B, Ouellette M. ABC Proteins of Leishmania. J Bioener Biomem. 2001; 33:469-74.
Haldar, Kumar Sen A, Roy P, Syamal Use of Antimony in the Treatment of Leishmaniasis: Current Status and Future Directions. Mol Biol Int. 2011; 10:1-23.
Higgins CF. ABC transporters: from microorganisms to man. Annu Rev Cell Biol. 1992; 8:67-/113.
Leandro C, Campino L. Leishmaniasis: efflux pumps and chemo resistance. Int J Antimicrob Agents. 2003; 22:352-/57.
Ouellette M, Fase-Fowler F, Borst P. The amplified H circle of methotrexate-resistant Leishmania tarentolae contains a novel Pglycoprotein gene. EMBO J. 1990; 9:1027-/33.
Fairlamb AH, Blackburn P, Ulrich P, Chait BT, Cerami A. Trypanothione: a novel bis (glutathionyl) spermidine cofactor for glutathione reductase in trypanosomatids. Science. 1985; 227:1485–7.
Mukhopadhyay R, Dey S, Xu N. Trypanothione overproduction and resistance to antimonials and arsenicals in Leishmania. Proc Natl Acad Sci USA. 1996; 93:10383–7.
Le´gare´ D, Papadopoulou B, Roy G. Efflux systems and increased trypanothione levels in arsenite-resistant Leishmania. Exp Parasitol. 1997; 87:275–82.
Esmaeili J, Mohebali M, Edrissian G. H, Rezayat S. M, Ghazi-Khansari M,. Charehdar S. Evaluation of miltefosine against Leishmana major (MRHO/IR/75/ER): in vitro and in vivo
studies. Acta Med Iranica. 2008; 46(3):191-196.
Pourmohammadi B, Motazedian MH, Handjani F, Hatam GH, Habibi S, Sarkari B. Glucantime efficacy in the treatment of zoonotic cutaneous leishmaniasis. Southeast Asian J Trop Med Public Health. 2011; 42:502-8.
Croft SL. Monitoring drug resistance in leishmaniasis. Trop Med Int Health. 2001; 6:899– 905.
Sundar S. Drug resistance in Indian visceral leishmaniasis. Trop Med Int Health. 2001; 6:849-854.
Lira R, Sundar S, Makharia A, Kenney R, Gam A, et al. Evidence that the high incidence of treatment failures in Indian Kala-Azar is due to the emergence of antimony-resistant strains of Leishmania donovani. J Infect Dis. 1999; 180:564– 567.
Mary C, Faraut F, Deniau M, Dereure J, Aoun K, Ranque S, et al. Frequency of Drug Resistance Gene Amplification in Clinical Leishmania Strains. Int Microbiol. 2010; 5:1-8.
Ouellette M, Papadopoulou B. Mechanisms of Drug Resistance in Leishmania. Parasitol Today. 1993; 9:150-53.
Alizadeh R, Hooshyar H, Bandehpor M, Arbabi M, Kazemi F, Talari A et al. Detection
of Drug Resistance Gene in Cutaneous Leishmaniasis by PCR in Some Endemic Areas of Iran. Iran Red Crescent Med J. 2011; 13(12):863-867.
Bagher Khadem E M, Mohebali M, Kazemirad E, Hajjaran H, Edrissian G, Mamishi S et
al. Downregulation of Calcineurin Gene Is Associated with Glucan-time®Resiatance in
Leishmania infantum. Iranian J Parasitol. 2013; 8(3):359-366.
Heather L. Callahan A, William L. Roberts B, Petrie M, Rainey b, Stephen M, Beverley A, The PGPA gene of Leishmania major mediates antimony (SblII) resistance by decreasing influx and not by increasing efflux. Mol Biochem Parasitol. 1994; 68:145-49.
Maharjan M, Singh S, Chatterjee M, Rentala Madhubala. Role of Aquaglyceroporin (AQP1) Gene and Drug Uptake in Antimony-resistant Clinical Isolates of Leishmania donovani. Am J Trop Med Hyg. 2008; 79:69–75.
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