فراوانی ژن پمپ های افلاکس PGP و MRPA در ایجاد مقاومت دارویی در ایزوله های بالینی لیشمانیا تروپیکا و لیشمانیا ماژور

نویسندگان

1 بیوتکنولوژی میکروبی، دانشگاه پیام نور تهران شرق، تهران، ایران

2 دانشکده بهداشت، دانشگاه علوم پزشکی قم، قم، ایران

چکیده
هدف‌ این تحقیق‌ شناسایی ‌ژن‌ پمپ‌های MRPA و PGP در ‌ایزوله‌های‌ بالینی ‌لیشمانیا است. از جمله‌ی ‌این ژن‌های‌ دخیل‌ در‌ مقاومت (MRPA) pgpa و (PGP) mdr1 هستند، که‌ محصول‌ آنها به‌ عنوان‌ انتقال‌دهنده وابسته ‌به ((ABC TransporterATP در ‌افلاکس ‌دارو‌ از ‌سیتوزول ‌به خارج غشا نقش دارند.‌ در این‌ راستا 40 داوطلب‌ مبتلا به‌ سالک ‌‌به طور‌ اتفاقی انتخاب گردیدند.‌ ابتدا آماستیگوت ها‌ با‌‌ میکروسکوپ نوری ‌بررسی گردیدند، سپس به‌ محیط ‌کشت دو فازی اختصاصیNNNتلقیح شدند.‌‎‌ DNA‌‌ ها به‌ روش فنل-کلروفرم ‌استخراج شدند و‌ با‌ پرایمرهای ‌اختصاصی ناحیه ITS عامل‌ ایجاد‌ زخم ‌تعیین ‌هویت‌ گردیدند. سپس فراوانی ‌دو ‌پمپ‌ دخیل ‌در‌ مقاومت‌ دارویی‌ به ‌کمک PCR بررسی‌ شد. در ‌این‌ مطالعه‌ ژن mdr1 که ‌پیش ‌از‌ این ‌در‌ سویه‌های ‌آزمایشگاهی ‌مقاوم‌‌‎‌ شده‌ به ‌ دارو‌ نشان ‌داده ‌شده بود، ‌نسبت به pgpa فراوانی‌ بیشتری ‌را دارا‌ بودند ‌که‌ یکی ‌از ‌دلایل ‌افزایش را می توان به‌ علت‌ حضور‌ پمپ MDR در‌سطح غشای‌ پلاسمایی اشاره‌ کرد‌، که‌ مواد‌‌ و دارو‌ را از‌‌ لایه های ‌‌داخلی‌ غشای دو ‌لایه ‌لیپیدی به ‌لایه های‌ خارجی انتقال‌ می‌دهد و‌ غلظت ‌دارو را‌ در‌ داخل‌ سلول ‌کم می‌کند و باعث مقاومت‌ دارویی ‌می‌شود در حالی‌که‌ پمپ MRPA در‌ غشا اندامک‌‌ سلولی است.


کلیدواژه‌ها


عنوان مقاله English

Frequency of PGP and MRPA efflux pump genes in drug resistance in clinical isolates of Leishmania tropica and L. major

نویسندگان English

Maede Parishan 1
Mahmoud Nateghi 2
1 Microbial Biotechnology, Payame Noor University of East Tehran, Tehran, Iran
2 Qom University of medical science, Health college, Qom, Iran
چکیده English

This study aimed to identify PGP and MRPA genes in clinical isolates of Leishmania. The genes of pgpa (MRPA) and mdr1 (PGP) are involved in the drug resistance, their products act as dependent transporters of ATP (ABC Transporter) in the reflux of drugs from the cytosol to the outer space of the cell. Hence, 40 volunteers with leishmaniasis were randomly selected. Firstly, Amastigotes were examined under a light microscope, then inoculated into NNN-specific biphasic culture medium. Deoxy ribonucleic acids were extracted by phenol-chloroform method and were determined by ITS-specific primers. Then the frequency of two pumps involved in "drug resistance" was investigated by PCR. In this study, the mdr1 gene, which had previously been shown to be present in the in vitro resistant strains, was shown to have a higher frequency of pgpas, which could be due to the presence of MDR. It transports the drug from the inner layers of the lipid bilayer membrane to the outer layers, reducing the concentration of the drug inside the cell and causing drug resistance, while the MRPA pump is in the membrane of the cell organelles.


کلیدواژه‌ها English

ABC transporter
cutaneous leishmaniasis
glucantim
mdr1 gene
pgpa gene
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  • تاریخ دریافت 18 خرداد 1405
  • تاریخ انتشار 18 خرداد 1405