vspВопросы современной педиатрииCurrent Pediatrics1682-55271682-5535Издательство «ПедиатрЪ»10.15690/vsp.v12i6.886vsp-268Research ArticleВ ПОМОЩЬ ВРАЧУA DOCTOR’S AIDСПЕКТР БЕЛКОВ, ПРОДУЦИРУЕМЫЙ ДЕРМАЛЬНЫМИ ФИБРОБЛАСТАМИ, У ДЕТЕЙ С БОЛЕЗНЬЮ КРОНАPROTEIN SPECTRUM PRODUCED BY THE DERMAL FIBROBLASTS IN CHILDREN WITH CROHN’S DISEASEКропотовВ. С.KropotovV. S.

Kropotov Vasiliy Sergeevich, Candidate of Biological Science, Senior Researcher at the Laboratory of Clinical Immunology and Cell Technologies

niidg@bk.ruКолесовС. А.KolesovS. A.niidg@bk.ruВасильеваЕ. А.Vasil'evaE. А.niidg@bk.ruМаянскаяИ. В.MayanskayaI. V.niidg@bk.ruАшкиназиВ. И.AshkinaziV. I.niidg@bk.ruНижегородский НИИ детской гастроэнтерологииРоссияNizhny Novgorod Scientific Research Institute of Pediatric GastroenterologyRussian FederationНижегородский НИИ детской гастроэнтерологиияРоссияNizhny Novgorod Scientific Research Institute of Pediatric GastroenterologyRussian Federation201307122013126Вопросы современной педиатрии120122Copyright © Кропотов В.С., Колесов С.А., Васильева Е.А., Маянская И.В., Ашкинази В.И., 20132013Кропотов В.С., Колесов С.А., Васильева Е.А., Маянская И.В., Ашкинази В.И.Kropotov V.S., Kolesov S.A., Vasil'eva E.А., Mayanskaya I.V., Ashkinazi V.I.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vsp.spr-journal.ru/jour/article/view/268

Представлены данные по исследованию продукции дермальными фибробластами спектра протеинов у условно здоровых детей и детей с болезнью Крона в ходе последовательных пассажей. Цель исследования: установить особенности белкового спектра супернатантов, полученных при многократном культивировании фибробластов в норме и при патологии. Пациенты и методы: объектом исследования были супернатанты 4 культур фибробластов, полученные из биоптатов кожи двух условно здоровых детей и двух детей с болезнью Крона в возрасте 14–17 лет. Оценку белкового спектра проводили на времяпролетном спектрометре Bruker Autoflex (США) методом масс-спектрометрического анализа MALDI-TOF, фенотипирование — на проточном цитофлуориметре FACS Cantu II (США). Результаты: методами статистического анализа в протеограммах были выделены 5 кластеров распределения протеинов по молекулярной массе — от 0,8 до 3,3 кДа, от 3,3 до 6,00 кДа, от 6 до 11 кДа, от 11 до 16,6 кДа и более 16,6 кДа. При сравнении протеограмм больных и условно здоровых детей обнаружены различия. Выводы: установлены определенные особенности протеомного профиля, характерные для супернатантов культур фибробластов детей с болезнью Крона. Зарегистрированные показатели протеомного спектра секреторных компонентов культур фибробластов могут быть использованы для характеристики их активности на различных сроках культивирования.

The results of the study on production of the protein spectrum by the dermal fibroblasts in relatively healthy children and children with Crohn’s disease during consecutive passages are shown in this article. Aim: to establish characteristics of protein spectrum of supernatants derived from the repeated cultivation of fibroblasts under normal and pathological conditions. Patients and methods: supernatants of 4 fibroblast cultures obtained from cutaneous tissue samples of 2 relatively healthy and 2 children with Crohn’s diseases aged from 14 to 17 years old were examined. The assessment of protein spectrum was performed by time-of-flight mass spectrometry MALDI-TOF (Bruker Autoflex, USA); phenotyping was performed by flaw cytometry assay (FACS Cantu II, USA). Results: 5 clusters of protein distribution according to the molecular weight were distinguished by the methods of statistical analysis:  from 0,8 to 3,3 kDa, from 3,3 to 6,0 kDa, from 6,0 to 11,0 kDa, from 11,0 to 16,6 kDa and >16,6 kDa. Proteinograms of patients with Crohn’s disease and relatively healthy children differ from each other. Conclusions: certain special features of proteomic profile, characterizing supernatants of fibroblastic cultures of children with Crohn’s disease were found. The established characteristics of proteomic spectrum of fibroblast secretory components can be used as marker of their activity at different stages of cultivation.

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