fa تغییر بیان ژن mexB در جدایه‌های سودوموناس آئروژینوزای مقاوم به سیپروفلوکساسین تحت تیمار با سیلیبینین انکپسوله در نانوذرات میکروبیولوژی Microbiology تحقیقی Research <p dir="RTL" style="text-align: justify;"><span style="font-family:times new roman;"><strong><span style="color:blue;">سابقه و هدف:</span></strong> <span style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">سیلیبینین (سیلیبین) یک</span></span></span> <span style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">ترکیب فعال سیلیمارین با فعالیت‌های ضدباکتریایی است. در جدایه‌های <em>سودوموناس آئروژینوزا</em> مقاوم به سیپروفلوکساسین افزایش بیان ژن </span></span></span><em><span dir="LTR" style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">mexB</span></span></span></em><span style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;"> گزارش شده است. هدف از این مطالعه، ارزیابی بیان ژن </span></span></span><em><span dir="LTR" style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">mexB</span></span></span></em> <span style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">در جدایه‌های <em>سودوموناس آئروژینوزای</em> تیمار شده و تیمار نشده با سیلیبینین بود.</span></span></span><br> <strong><span style="color:blue;"><span style="font-size:10.0pt;">مواد و روش&shy;ها:</span></span></strong> <span style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">در این مطالعه توصیفی- تحلیلی، جدایه های <em>سودوموناس آئروژینوزا </em>از بیمارستان ها و آزمایشگاههای استان گیلان تهیه شد. بعد از تعیین حساسیت به چند آنتی بیوتیک (دیسک دیفیوژن و </span></span></span><span dir="LTR" style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">MIC</span></span></span><span style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">)، 5 جدایه <em>سودوموناس آئروژینوزای </em>مقاوم تحت تیمار با سیپروفلوکساسین (1/2</span></span></span><span dir="LTR" style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">MIC</span></span></span><span style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">) به تنهایی (گروه 1) و در ترکیب با سیلیبینین انکپسوله در میسل (نانوذرات) (گروه 2) قرار گرفت. بعد از 24 ساعت، استخراج </span></span></span><span dir="LTR" style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">RNA</span></span></span><span style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;"> و سنتز </span></span></span><span dir="LTR" style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">cDNA</span></span></span><span style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;"> در گروه 1 و 2 صورت گرفت و بیان ژن </span></span></span><em><span dir="LTR" style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">mexB</span></span></span></em><span style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;"> به روش </span></span></span><span dir="LTR" style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">Realtime-PCR</span></span></span><span style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;"> کمی (</span></span></span><span dir="LTR" style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">Q-RT-PCR</span></span></span><span style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">) و معادله </span></span></span><span dir="LTR" style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">2^{-&Delta;&Delta;CT}</span></span></span><span style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;"> ارزیابی گردید.</span></span></span><br> <strong><span style="color:blue;"><span style="font-size:10.0pt;">یافته&shy; ها:</span></span></strong> <span style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">در این مطالعه از 69 جدایه <em>سودوموناس آئروژینوزا</em>، 33/33% به روش دیسک دیفیوژن و 37/68%&nbsp; به روش </span></span></span><span dir="LTR" style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">MIC</span></span></span><span style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;"> مقاوم به سیپروفلوکساسین تشخیص داده شدند. نتایج نشان داد که سیلیبینین انکپسوله در نانوذرات (</span></span></span><span dir="LTR" style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">&micro;g/ml</span></span></span><span style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">400)، مرگ را در جدایه‌های مقاوم تحت تیمار با سیپروفلوکساسین (1/2</span></span></span><span dir="LTR" style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">MIC</span></span></span><span style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">) تا 50% القاء می‌کند. در سلولهای تحت تیمار با سیلیبینین و سیپروفلوکساسین کاهش بیان ژن </span></span></span><em><span dir="LTR" style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">mexB</span></span></span></em> <span style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">در مقایسه با سلولهای تحت تیمار با سیپروفلوکساسین به تنهایی آشکار شد.</span></span></span><br> <strong><span style="color:blue;"><span style="font-size:10.0pt;">نتیجه&shy; گیری:</span></span></strong> <span style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">به نظر می رسد سیلیبینین از طریق کاهش بیان ژنهای درگیر در سیستم های افلاکس پمپ نظیر </span></span></span><span dir="LTR" style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;">mexB</span></span></span><span style="background-image:initial;background-position:initial;background-size:initial;background-repeat:initial;background-attachment:initial;background-origin:initial;background-clip:initial;"><span style="color:black;"><span style="font-size:10pt;"> باعث افزایش تأثیر سیپروفلوکساسین در مهار رشد سودوموناس آئروژینوزا شود</span></span></span><span style="font-size:10.0pt;">.</span></span><br> &nbsp;</p> <p dir="RTL" style="text-align: justify;"></p> <p style="text-align: justify;"><span style="font-family:times new roman;"><span style="color:#0000FF;"><strong>BACKGROUND AND AIM:</strong></span> Silibinin (<a href="https://www.ncbi.nlm.nih.gov/mesh/67440975">silybin</a>) is an active component of silymarin with anti-bacterial activities. In drug resistant isolates of <em>Pseudomonas aeruginosa</em> has been reported overexpression of <em>mexB</em> gene. The aim of this study was evaluation of <em>mexB</em> gene expression in silibinin treated and untreated isolates of <em>Pseudomonas aeruginosa</em>.<br> <span style="color:#0000FF;"><strong>METHODS:</strong> </span>In this descriptive analytical study,<em> Pseudomonas aeruginosa</em> isolates were obtained from hospitals and laboratories of Guilan province. After determining several antibiotics susceptibility (disc diffusion and MIC), 5 ciprofloxacin resistant isolates of <em>Pseudomonas aeruginosa</em> were treated by ciprofloxacin (1/2MIC) only (group1) and in the combination with silibinin-encapsulated&nbsp;micelles (nanoparticles) (group2). After 24h, RNA extraction and cDNA synthesis was performed in group1 and group2 and <em>mexB</em> gene expression was evaluated by quantitative-realtime PCR (Q-RT-PCR) and 2^{-&Delta;&Delta;CT} equation.<br> <span style="color:#0000FF;"><strong>FINDINGS:</strong></span> In this study from 69 isolates, 33.33% by disc diffusion test and 37.86% by MIC test were determined ciprofloxacin resistant. Our analysis showed that silibinin -encapsulated&nbsp;nanoparticles (400&micro;g/ml) induced death up t0 50% in ciprofloxacin (1/2MIC) treated resistant isolates during 24h. In treated cells with silibinin and ciprofloxacin revealed downregulation of <em>mexB</em> gene compared to treated cells with ciprofloxacin alone.<br> <span style="color:#0000FF;"><strong>CONCLUSION:</strong></span> It seems that silibinin is cause of increasing ciprofloxacin effect on inhibition of growth of <em>Pseudomonas aeruginosa</em> through decreasing expression of genes implicated in efflux pomp systems such as <em>mexB</em>.</span><br> &nbsp;</p> <p style="text-align: justify;"></p><p style="text-align: justify;"></p> سیپروفلوکساسین, mexB, نانوذرات, سودوموناس آئروژینوزا, سیلیبین Ciprofloxacin, mexB, Nanoparticles, Pseudomonas aeruginosa, Realtime PCR, Silybin 42 49 http://jbums.org/browse.php?a_code=A-10-3191-1&slc_lang=fa&sid=1 Z Ahmadi Roudbaraki زهرا احمدی رودبارکی zahra.ahmadi1212@gmail.com 100319475328460025281 100319475328460025281 No Islamic Azad University, Rasht دانشگاه آزاد اسلامی ، رشت N Ranji نجمه رنجی najmehranji@gmail.com 100319475328460025282 100319475328460025282 Yes Islamic Azad University دانشگاه آزاد اسلامی ، رشت B Soltani Tehrani بهرام سلطانی تهرانی soltani@gums.ac.ir 100319475328460025283 100319475328460025283 No Guilan University of Medical Sciences, دانشگاه علوم پزشکی گیلان، رشت، ایران.