شناسایی مولکولی ژن پروتئین فعال کننده نوتروفیل هلیکوباکتر پیلوری موثر در کنترل آسم آلرژیک به روش PCR
محورهای موضوعی : ایمنی شناسی
علیرضا خالقی خرمی
1
,
ساناز مهمازی
2
,
رسول شکری
3
,
سید شمس الدین اطهاری
4
1 - گروه میکروبیولوژی، دانشکده علوم پایه، واحد زنجان، دانشگاه آزاد اسلامی، زنجان، ایران
2 - گروه میکروبیولوژی، دانشکده علوم پایه، واحد زنجان، دانشگاه آزاد اسلامی، زنجان، ایران
3 - گروه میکروبیولوژی، دانشکده علوم پایه، واحد زنجان، دانشگاه آزاد اسلامی، زنجان، ایران
4 - گروه ایمونولوژی، دانشکده پزشکی، دانشگاه علوم پزشکی زنجان، زنجان، ایران
کلید واژه: هلیکوباکتر پیلوری, آسم آلرژیک, پروتئین فعال کننده نوتروفیل,
چکیده مقاله :
پروتئین فعال کننده نوتروفیل هلیکوباکتر پیلوری (HP-NAP) یکی از پروتئین های مهم هلیکوباکتر پیلوری (H. pylori) است HP-NAP.دارای خواص تعدیل کننده ایمنی است و پاسخ های آلرژیک را از Th2 به Th1 هدایت می کند. هدف از این مطالعه شناسایی مولکولی ژن HP-NAP بومی ایران است. سویه بومی هلیکوباکتر پیلوری از انستیتو پاستور خریداری شد. آزمایش اوره آز اختصاصی، رنگ‌آمیزی گرم و مشاهده میکروسکوپی انجام شد. برای شناسایی HP-NAP در باکتری های بومی از روش PCR استفاده شد. در نهایت، محصول PCR برای تشخیص ژن HP-NAP الکتروفورز شد. پس از رنگ‌آمیزی باکتریایی و نیز مشاهده میکروسکوپی، باکتری‌های گرم منفی خمیده مشاهده شدند. آزمایش اوره آز تغییر در معرف اوره (فنل قرمز) را نشان داد که نشان دهنده هلیکوباکتر پیلوری است. آزمایش PCR و الکتروفورز یک باند 260 جفت باز مربوط به ژن HP-NAP را در مقایسه با نردبان (لدر) 100 نشان داد. با توجه به مطالعه انجام شده و تایید وجود این پروتئین در ساختار باکتری های بومی و همچنین مطالعات قبلی در در مناطق مختلف جهان، با مشاهده اثرات این پروتئین به عنوان تعدیل کننده ایمنی سیستم ایمنی، می توان امیدوار بود که در آینده از این پروتئین به عنوان وسیله ای بالقوه برای کنترل و درمان بیماری های آلرژیک به ویژه آسم آلرژیک استفاده شود.
Helicobacter pylori neutrophil activating protein (HP-NAP) is one of the important proteins of Helicobacter pylori (H. pylori). HP-NAP has immunomodulatory properties and directs allergic responses from Th2 to Th1. The aim of this study is the molecular identification of the HP-NAP gene native to Iran. The native strain of H. pylori was purchased from the Pasteur Institute. A specific urease test was performed, along with gram staining and microscopic observation. To identify HP-NAP in native bacteria, the PCR method was used. Finally, the PCR product was electrophoresed to detect the HP-NAP gene. After bacterial staining and microscopic observation, bent gram-negative bacteria were observed. The urease test showed a change in the urea reagent (phenol red), indicating H. pylori. The PCR test and electrophoresis revealed a 260 bp band related to the HP-NAP gene, compared with ladder 100. According to the study conducted and the confirmation of the presence of this protein in the structure of native bacteria, as well as previous studies in different regions of the world, observing the effects of this protein as an immunomodulator of the immune system, we can hope that in the future this protein will be used as a potential means to control and treat allergic diseases, especially allergic asthma.
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