همبستگی اینترلوکین-۶ به عنوان یک آزمون ایمنی‌شناختی با دیروفیلاریازیس نهفته در سگ (COD)

نوع مقاله : مقاله پژوهشی

نویسنده

دانشگاه فنی میانه، مؤسسه فناوری پزشکی/ بغداد، رشته فناوری تولید پروتز و ارتز، بغداد، عراق.

چکیده

هدف: دیروفیلاریازیس نهفته در سگ (COD) یک شکل تحت‌بالینی از عفونت Dirofilaria immitis است که در آن سگ‌ها انگل را بدون نشان دادن علائم بالینی حمل می‌کنند. هنوز مشخص نیست که سیتوکین‌های ایمنی، به‌ویژه IL-6 و IL-10، چگونه بر پاسخ ایمنی در طی COD تأثیر می‌گذارند. اینترلوکین ۶ (IL-6) یک سیتوکین پیش‌التهابی است. هدف این مطالعه بررسی همبستگی اینترلوکین ۶ به عنوان یک آزمون ایمنی‌شناختی در طی COD بود.
مواد و روش‌ها: در مجموع ۵۶ سگ نر و ۴۴ سگ ماده (۱۰۰ حیوان) با دامنه سنی ۱ تا ۱۰ سال مورد بررسی قرار گرفتند. اصول آسپتیک در زمان جمع‌آوری خون و نمونه‌برداری به‌طور دقیق رعایت شد. برای بررسی وجود لاروهای میکروفیلاریا، اسمیر خون محیطی از خون تازه تهیه گردید. از ۱۰۰ نمونه آزمایش‌شده، ۵۰ سگ سالم به عنوان گروه کنترل و ۵۰ سگ به‌طور طبیعی به Dirofilaria immitis آلوده بودند. برای بررسی بیان ژن IL-6 از qPCR استفاده شد. همچنین ژن TBP به عنوان ژن مرجع به کار رفت. برای تجزیه و تحلیل داده‌ها از نرم‌افزار SPSS نسخه ۲۹ استفاده شد.
نتایج: میانگین سطح پلاسمایی IL-6 در گروه کنترل و گروه دارای دیروفیلاریازیس نهفته به ترتیب 6/7 ± 8/56 پیکوگرم بر میلی‌لیتر و 2/10 ± 5/124 پیکوگرم بر میلی‌لیتر بود. نتایج منحنی‌های PCR در زمان واقعی و الکتروفورز ژل آگارز محصولات PCR نشان داد که ژن IL-6 در سگ‌های مورد بررسی تکثیر و بیان می‌شود. یک باند ۱۱۴ جفت‌بازی برای این ژن و یک باند ۱۸۴ جفت‌بازی برای ژن TBP در تمامی نمونه‌ها مشاهده شد. میانگین تغییر بیان (fold-change) ژن IL-6 در گروه کنترل 2/0 ± 1 و در سگ‌های بیمار 9/0 ± 2/4 بود. در نمونه‌های مورد مطالعه، یک ضریب همبستگی منفی (r = − 0.34) برای همبستگی پیرسون بین سطوح IL-6 و بار انگل مشاهده شد (p < 0.05).
نتیجه‌گیری: نتایج این مطالعه نشان داد که هم سطح پلاسمایی و هم بیان ژنی IL-6 در سگ‌های مبتلا به دیروفیلاریازیس نهفته افزایش می‌یابد. علاوه‌براین، این افزایش باعث تغییر در بار انگل شده و با آن همبستگی داشت. بنابراین، IL-6 می‌تواند به عنوان یک نشانگر التهابی بالقوه در مدیریت و بررسی بالینی این عفونت مورد استفاده قرار گیرد. با این حال، پیش از هرگونه اقدام، مطالعات طولی با حجم نمونه بزرگ‌تر و با استفاده همزمان از مجموعه گسترده‌تری از نشانگرهای زیستی برای تأیید این نقش لازم است.

کلیدواژه‌ها


عنوان مقاله [English]

The correlation of interleukin-6 as an immunological test with canine occult dirofilariasis

نویسنده [English]

  • Amal Hasan Atiyah
Middle Technical University, Institute of Medical Technology/ Baghdad Prosthesis and Orthosis Production Techniques, Baghdad, Iraq.
چکیده [English]

Objective
Canine occult dirofilariasis (COD) is a subclinical form of Dirofilaria immitis infection, where dogs carry the parasite without exhibiting clinical symptoms. It is unclear how immune cytokines, especially IL-6 and IL-10, affect the immunological response during COD. IL-6 is pro-inflammatory. This study aimed to investigate the correlation of interleukin-6 as an immunological test during COD.

Materials and methods
A total of 56 male and 44 female dogs (total number 100) ranging in age from 1 to 10 years were studied. Aseptic principles were strictly observed for blood collection and sample collection. To examine the presence of microfilaria larvae, peripheral blood smears were prepared from fresh blood. Of the 100 samples tested, 50 were healthy dogs and were used as a control group, and 50 dogs were naturally infected with Dirofilaria immitis. qPCR was used to examine IL-6 gene expression. In addition, the TBP gene was used as a reference gene. SPSS version 29 software was used to analyze the data.

Results
The mean plasma IL-6 levels for the control group and the group with occult dirofilariasis were 56.8 ± 7.6 pg/mL and 124.5 ± 10.2 pg/mL, respectively. The results of the real-time PCR curves and the agarose gel electrophoresis of the PCR products showed that the IL-6 gene is amplified and expressed in the studied dogs. A 114 bp band was observed for this gene and a 184 bp band was observed for the TBP gene in all samples. The mean fold-change of IL-6 gene for the control group was 1.0 ± 0.2 and for the diseased dogs was 4.2 ± 0.9. In the studied samples, a negative correlation coefficient (r = −0.34) was observed for the Pearson correlation between IL-6 levels and parasite load (p < 0.05).

Conclusion
The results of this study showed that both plasma levels and gene expression of IL-6 are increased in dogs with latent dirofilariasis. In addition, this increase caused changes in parasite load and was correlated with them. Therefore, IL-6 can be used as a potential inflammatory marker in the management and clinical investigation of this infection. However, before any action, longitudinal studies with larger samples are needed to confirm this role, while simultaneously benefiting from a broader set of biomarkers in the study.

کلیدواژه‌ها [English]

  • canine occult dirofilariasis (COD)
  • dirofilaria immitis infection
  • interleukin-6
  • immunological
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