Journal of Mazandaran University of Medical Sciences
مجله دانشگاه علوم پزشکی مازندران
J Mazandaran Univ Med Sci
Medical Sciences
http://jmums.mazums.ac.ir
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1393
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2014
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بررسی اثر فاکتور رشد انسولینی در روند ترمیم زخم های حاد پوستی در رت های دیابتی شده به وسیله استرپتوزوتوسین
Effect of IGF-1 Gene Therapy in Acute Cutaneous Wound Healing in Streptozotocin-Induced Diabetic Rats
پژوهشي-کامل
Research(Original)
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<p style="text-align: justify line-height: 200% direction: rtl unicode-bidi: embed" dir="rtl" class="MsoNormal"><span lang="AR-SA" style="font-size: 16pt line-height: 200% font-family: "B Koodak" color: black">چکیده</span></p>
<p style="text-align: justify text-indent: 17pt line-height: 200% direction: rtl unicode-bidi: embed" dir="rtl" class="MsoNormal"><span lang="FA" style="line-height: 200% font-family: "B Koodak" color: black">سابقه</span><span lang="AR-SA" style="line-height: 200% font-family: "B Koodak" color: black"> و هدف:</span><span lang="AR-SA" style="line-height: 200% font-family: "B Zar""> </span><span lang="FA" style="line-height: 200% font-family: "B Zar"">تاخیر در ترمیم زخم افراد دیابتی یک پدیده شایع است که
پاتولوژی آن کم</span><span lang="AR-SA" style="line-height: 200% font-family: "B Zar""></span><span lang="FA" style="line-height: 200% font-family: "B Zar"">تر شناخته شده
است. انواع سلول</span><span lang="AR-SA" style="line-height: 200% font-family: "B Zar""></span><span lang="FA" style="line-height: 200% font-family: "B Zar"">های پوستی،
ماتریکس خارج سلولی و انواع فاکتورهای رشد در ترمیم زخم دخالت دارند. تحقیقات
زیادی در خصوص اثرات ژن درمانی و سلول درمانی در بهبود زخم انجام شده است. در این
مطالعه تجربی، اثرات انتقال سلول فیبروبلاست ترنسفکته شده با ژن </span><span style="font-size: 10pt line-height: 200%" dir="ltr">IGF-1</span><span dir="rtl"></span><span lang="FA" style="line-height: 200% font-family: "B Zar""><span dir="rtl"></span> بر روی زخم</span><span lang="AR-SA" style="line-height: 200% font-family: "B Zar""></span><span lang="FA" style="line-height: 200% font-family: "B Zar"">های دیابتیک در رت مورد بررسی قرار گرفت.</span></p>
<p style="text-align: justify text-indent: 17pt line-height: 200% direction: rtl unicode-bidi: embed" dir="rtl" class="MsoNormal"><span lang="AR-SA" style="line-height: 200% font-family: "B Koodak" color: black">مواد و روش</span><span lang="FA" style="line-height: 200% font-family: "B Zar" letter-spacing: -0.2pt"></span><span lang="AR-SA" style="line-height: 200% font-family: "B Koodak" color: black">ها:</span><span lang="AR-SA" style="line-height: 200%"> </span><span lang="FA" style="line-height: 200% font-family: "B Zar"">در مطالعه
تجربی حاضر، دیابت در موش</span><span lang="AR-SA" style="line-height: 200% font-family: "B Zar""></span><span lang="FA" style="line-height: 200% font-family: "B Zar"">های صحرایی با
</span><span lang="AR-SA" style="line-height: 200% font-family: "B Zar"">استرپتوزوتوسین</span><span lang="FA" style="line-height: 200% font-family: "B Zar""> القا شد.
فیبروبلاست</span><span lang="AR-SA" style="line-height: 200% font-family: "B Zar""></span><span lang="FA" style="line-height: 200% font-family: "B Zar"">ها کشت داده شدند و
با </span><span style="font-size: 10pt line-height: 200%" dir="ltr">IGF-1</span><span dir="rtl"></span><span lang="FA" style="line-height: 200% font-family: "B Zar""><span dir="rtl"></span> ترنسفکته شدند.
از لیپوفکتامین 2000 به عنوان معرف ترنسفکشن استفاده شد. میزان بیان </span><span style="font-size: 10pt line-height: 200%" dir="ltr">IGF-1</span><span dir="rtl"></span><span style="line-height: 200% font-family: "B Zar""><span dir="rtl"></span> <span lang="FA">ترنس</span></span><span lang="AR-SA" style="line-height: 200% font-family: "B Zar""></span><span lang="FA" style="line-height: 200% font-family: "B Zar"">ژن شده به وسیله الایزا اندازه</span><span lang="AR-SA" style="line-height: 200% font-family: "B Zar""></span><span lang="FA" style="line-height: 200% font-family: "B Zar"">گیری شد. جهت مطالعه در محیط </span><span style="font-size: 10pt line-height: 200%" dir="ltr">In Vivo</span><span dir="rtl"></span><span style="line-height: 200% font-family: "B Zar""><span dir="rtl"></span> <span lang="FA">حیوانات (با وزن
200-170 گرم) به طور تصادفی به سه گروه (5 رت/گروه) تقسیم شدند و زخم</span></span><span lang="AR-SA" style="line-height: 200% font-family: "B Zar""></span><span lang="FA" style="line-height: 200% font-family: "B Zar"">هایی با ضخامت کامل پوست در ناحیه پشت گردن ایجاد شد.
سوسپانسیونی از سلول</span><span lang="AR-SA" style="line-height: 200% font-family: "B Zar""></span><span lang="FA" style="line-height: 200% font-family: "B Zar"">های
فیبروبلاست ترنسفکته و ترنسفکته نشده به لبه زخم تزریق شد و با زخم</span><span lang="AR-SA" style="line-height: 200% font-family: "B Zar""></span><span lang="FA" style="line-height: 200% font-family: "B Zar"">های درمان شده با نرمال سالین مقایسه گردید. برای آزمایشات
میکروسکوپی، بیوپسی در روز 8 انجام شد.<span style="color: black"></span></span></p>
<p style="text-align: justify text-indent: 17pt line-height: 200% direction: rtl unicode-bidi: embed" dir="rtl" class="MsoNormal"><span lang="AR-SA" style="line-height: 200% font-family: "B Koodak" color: black">یافته</span><span lang="FA" style="line-height: 200% font-family: "B Zar" letter-spacing: -0.2pt"></span><span lang="AR-SA" style="line-height: 200% font-family: "B Koodak" color: black">ها:</span><span lang="AR-SA" style="font-size: 14pt line-height: 200% font-family: "B Lotus""> </span><span lang="FA" style="line-height: 200% font-family: "B Zar"">در محیط آزمایشگاهی، حداکثر بیان</span><span style="font-size: 10pt line-height: 200%" dir="ltr">IGF-1</span><span dir="rtl"></span><span style="line-height: 200% font-family: "B Zar""><span dir="rtl"></span> <span lang="FA">در سلول</span></span><span lang="AR-SA" style="line-height: 200% font-family: "B Zar""></span><span lang="FA" style="line-height: 200% font-family: "B Zar"">های فیبروبلاست ترنسفکته (95/96 پیکوگرم/ میکرولیتر)، 24
ساعت بعد از ترنسفر ژن بود. همچنین در محیط </span><span style="font-size: 10pt line-height: 200%" dir="ltr">In Vivo</span><span dir="rtl"></span><span lang="FA" style="line-height: 200% font-family: "B Zar""><span dir="rtl"></span>، ژن درمانی باعث افزایش فیبروبلاست</span><span lang="AR-SA" style="line-height: 200% font-family: "B Zar""></span><span lang="FA" style="line-height: 200% font-family: "B Zar"">ها و کراتینوسیت</span><span lang="AR-SA" style="line-height: 200% font-family: "B Zar""></span><span lang="FA" style="line-height: 200% font-family: "B Zar"">ها در فرایند ترمیم زخم شد. بافت گرانوله تشکیل شده در زخم</span><span lang="AR-SA" style="line-height: 200% font-family: "B Zar""></span><span lang="FA" style="line-height: 200% font-family: "B Zar"">های ترمیم شده با فیبروبلاست ترنسفکته و فیبروبلاست
ترنسفکته نشده در مقایسه با گروه کنترل سازمان یافته تر بود.<span style="color: black"></span></span></p>
<p style="text-align: justify text-indent: 17pt line-height: 200% direction: rtl unicode-bidi: embed" dir="rtl" class="MsoNormal"><span lang="AR-SA" style="line-height: 200% font-family: "B Koodak" color: black">استنتاج:</span><span lang="AR-SA" style="line-height: 200%"> </span><span class="hps"><span lang="FA" style="line-height: 200% font-family: "B Zar"">این مطالعه نشان داد که با بهینه</span></span><span lang="AR-SA" style="line-height: 200% font-family: "B Zar""></span><span class="hps"><span lang="FA" style="line-height: 200% font-family: "B Zar"">سازی انتقال ژن</span></span><span lang="FA" style="line-height: 200% font-family: "B Zar"">، <span class="hps">بیان</span></span><span class="hps"><span style="font-size: 10pt line-height: 200%" dir="ltr">IGF-1</span></span><span dir="rtl"></span><span class="hps"><span style="line-height: 200% font-family: "B Zar""><span dir="rtl"></span>
<span lang="FA">افزایش یافت و انتقال سلول فیبروبلاست ترنسفکته شده به محیط زخم</span></span></span><span lang="FA" style="line-height: 200% font-family: "B Zar"">، <span class="hps">دارای اثرات قابل توجهی در بهبود زخم</span></span><span lang="AR-SA" style="line-height: 200% font-family: "B Zar""></span><span class="hps"><span lang="FA" style="line-height: 200% font-family: "B Zar"">های حاد دیابتی در موش</span></span><span lang="AR-SA" style="line-height: 200% font-family: "B Zar""></span><span class="hps"><span lang="FA" style="line-height: 200% font-family: "B Zar"">های صحرایی بود.</span></span><span lang="FA" style="font-family: "B Zar" color: black"></span></p>
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Abstract
Background and purpose: Delayed wound healing is a common complication of diabetes mellitus. However, less is known about the cause of this pathology. Types of skin cells, extracellular matrix and variety of growth factors are involved in wound healing. The use of recombinant growth factors in researches and production of skin substitutes are still a challenge. Much research has been done on the effects of gene therapy and cell therapy in wound healing. This experimental study assessed the effect of insulin-like growth factor (IGF-1) over expression in combination with fibroblast cells therapy to diabetic wounds in rats.
Material and methods: Diabetes was induced in rats using Streptozotocin. Fibroblasts were cultured and transfected with IGF-1. Lipofectamine 2000 was used as reagent transfection and transgene expression levels were measured by ELIZA. To study the in vivo, rats (weighing 170-200g) were randomly divided into three groups (5/group) and full-thickness wounds were created on the dorsum region. Suspensions of transfected fibroblast and native fibroblast cells were injected into the wound and were compared with the wounds treated with normal saline. For microscopic examination, biopsy was performed on day 8.
Results: In in vitro, the maximum expression of IGF-1 in transfected fibroblast cells
(96.95 pg/ml) was 24 h after gene transfer. In vivo, IGF-1 gene therapy increased the number of fibroblast and keratinocyte cells in wound healing process. Granulation tissue formation in transfected fibroblast and native fibroblast cells groups compared with the normal saline group were found to be more organized.
Conclusion: The expression of IGF-1 increased by optimization of gene transfer. High concentrations of IGF-1, in combination with cell therapy, had a significant effect on delayed wound healing in diabetic rats.
فاکتور رشد انسولینی, ژن درمانی, دیابتی, ترمیم زخم
IGF-1, gene therapy, Diabetic, wound healing
1
11
http://jmums.mazums.ac.ir/browse.php?a_code=A-10-1269-8&slc_lang=fa&sid=1
Fereshteh Beygom
Talebpour Amiri
فرشته بیگم
طالب پور امیری
100319475328460038420
100319475328460038420
No
Assistant Professor, Department of Anatomy, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran
استادیار، گروه آناتومی، دانشکده پزشکی، دانشگاه علوم پزشکی مازندران، ساری، ایران
Mahnaz
Mahmoudirad
مهناز
محمودی راد
mahnazrad@gmail.com
100319475328460038421
100319475328460038421
Yes
Associate Professor, Skin Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
دانشیار، مرکز تحقیقات پوست، دانشگاه علوم پزشکی شهید بهشتی، تهران، ایران
Mehri
Mirhoseini
مهری
میر حسینی
100319475328460038422
100319475328460038422
No
Assistant Professor, Amol Faculty of Nursing & Midwifery, Mazandaran University of Medical Sciences, Sari, Iran
استادیار، دانشکده پرستاری و مامایی حضرت زینب آمل، دانشگاه علوم پزشکی مازندران، ساری، ایران
Mansoureh
Mirzaei
منصوره
میرزایی
100319475328460038423
100319475328460038423
No
MSc in Anatomy, Mazandaran University of Medical Sciences, Sari, Iran
کارشناسی ارشد آناتومی، دانشگاه علوم پزشکی مازندران، ساری، ایران