mimmunМедицинская иммунологияMedical Immunology (Russia)1563-06252313-741XSPb RAACI10.15789/1563-0625-2013-6-525-534mimmun-676Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИORIGINAL ARTICLESОПРЕДЕЛЕНИЕ мРНК АНГИОГЕННЫХ И АНГИОСТАТИЧЕСКИХ ХЕМОКИНОВ И ИХ РЕЦЕПТОРОВ В СИНОВИАЛЬНОЙ ОБОЛОЧКЕ МЕТОДОМ КОЛИЧЕСТВЕННОЙ ПЦР В РЕАЛЬНОМ ВРЕМЕНИANALYSIS OF EXPRESSION OF ANGIOGENIC AND ANGIOSTATIC CHEMOKINES AND THEIR RECEPTORS IN SYNOVIAL TISSUE BY QUANTITATIVE REAL-TIME PCRЖебрунД. А.ZhebrunD. A.

младший научный сотрудник

197341, ул. Аккуратова, 2. Тел.: 8 (812) 702-37-77

Research Associate, Laboratory of Immunology

197341, Russian Federation, St. Petersburg, Akkuratova str., 2. Phone: 7 (812) 702-37-77

zhebrun@almazovcentre.ruМаслянскийА. Л.MaslyanskyA. L.

к.м.н., старший научный сотрудник НИЛ ревматологии

PhD (Medicine), Senior Research Associate, Laboratory of Rheumatology Department

zhebrun@almazovcentre.ruТитовА. Г.TitovA. G.

к.м.н., врач-травматолог отделения ревматологии

PhD (Medicine), Rheumatology Department

zhebrun@almazovcentre.ruПатрухинА. П.PatruhinA. P.

врач-травматолог

Clinical Traumatologist

zhebrun@almazovcentre.ruКостареваА. А.KostarevaA. A.

к.м.н., директор института Молекулярной биологии и генетики

PhD (Medicine) Chief

zhebrun@almazovcentre.ruГольцеваИ. С.GoltsevaI. S.

студенка-магистр

Master Student

zhebrun@almazovcentre.ruТотолянАрег А.TotolianAreg A.

д.м.н., профессор, член-корр. РАМН, заместитель директора по научной работе; заведующий НИЛ иммунологии

PhD, MD (Medicine), Professor, Corresponding Member, Deputy Director, Head, Laboratory of Immunology

zhebrun@almazovcentre.ruФБУН «НИИ эпидемиологии и микробиологии имени Пастера», Санкт-Петербург; ФГБУ «ФЦСКЭ имени В.А. Алмазова» Минздрава России, Санкт-ПетербургРоссияSt. Petersburg Pasteur Institute of Epidemiology and Microbiology, St. Petersburg; Almazov Federal Heart, Blood and Endocrinology Centre, St. PetersburgRussian FederationФГБУ «ФЦСКЭ имени В.А. Алмазова» Минздрава России, Санкт-ПетербургРоссияAlmazov Federal Heart, Blood and Endocrinology Centre, St. PetersburgRussian FederationФБУН «НИИ эпидемиологии и микробиологии имени Пастера», Санкт-ПетербургРоссияSt. Petersburg Pasteur Institute of Epidemiology and Microbiology, St. PetersburgRussian FederationГОУ ВПО «Санкт-Петербургский государственный технологический институт», Санкт-ПетербургРоссияSt. Petersburg State Technological Institute, St. PetersburgRussian Federation201326072014156525534Copyright © Жебрун Д.А., Маслянский А.Л., Титов А.Г., Патрухин А.П., Костарева А.А., Гольцева И.С., Тотолян А.А., 20142014Жебрун Д.А., Маслянский А.Л., Титов А.Г., Патрухин А.П., Костарева А.А., Гольцева И.С., Тотолян А.А.Zhebrun D.A., Maslyansky A.L., Titov A.G., Patruhin A.P., Kostareva A.A., Goltseva I.S., Totolian A.A.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.mimmun.ru/mimmun/article/view/676

Резюме. В настоящее время в иммунопатогенезе различных заболеваний суставов большая роль отводится хемокинам. При обследовании больных с патологией суставов (ревматоидный артрит, остеоартроз, псориатический артрит и др.), помимо периферической крови, основным биоматериалом является синовиальная жидкость. В то же время не меньший интерес представляет исследование синовиальной оболочки (СО), в которой развиваются основные патологические процессы, и, соответственно, анализ хемокинов и их рецепторов в СО может предоставить более полную информацию об иммунопатогенезе заболеваний суставов различной этиологии. В данной работе была проведена оптимизация методики количественного определения экспрессии мРНК ангиогенных и ангиостатических хемокинов и их рецепторов в СО с помощью ПЦР в реальном времени с обратной транскрипцией. Подробно описаны все этапы проведения исследования, начиная от взятия биоматериала, заканчивая анализом результатов ПЦР. Приведены границы нормальных значений изучаемых показателей по результатам обследования лиц с травмой суставов в анамнезе и с отсутствием на момент обследования каких-либо признаков внутри- и внесуставного воспаления. Показано, что данный метод может быть информативен для сравнения внутрисуставной экспрессии хемокинов и рецепторов между группами и может использоваться для изучения роли хемокинов в развитии соответствующих иммунопатологических процессов.

Abstract. Chemokines are generally believed to play an important role in immunopathogenesis of different joint diseases. Synovial fluid is the primary bio-material, along with peripheral blood, for evaluation of patients with joint pathology (rheumatoid arthritis, osteoarthritis, psoriatic arthritis, etc). However, analysis of synovial membranes is also of importance, since the main pathologic processes evolve at this site. Analysis of chemokines and their receptors in synovium may provide more complete information about immunopathogenesis of articular disease of different etiologies. We carried out optimization procedures for real-time reverse-transcription PCR analysis, in order to quantify specific mRNA expression of angiogenic and angiostatic chemokines and their receptors in synovium. All the stages, from sampling the biological material and until analysis of results are described in details. Normal ranges for mRNA expression rates are provided, as based on examination of subjects with previous traumatic joint injury, without any clinical signs of current systemic or local inflammation. We demonstrate that this method can be informative for comparing intra-articular expression of chemokines and their receptors between different clinical groups. It can also be used to investigate the role of chemokines in appropriate immunopathological events.

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The authors declare that there are no conflicts of interest present.