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Современные технологии визуализации тучных клеток для биологии и медицины (обзор)

Современные технологии визуализации тучных клеток для биологии и медицины (обзор)

И.П. Григорьев, Д.Э. Коржевский
Ключевые слова: тучные клетки; гистохимия; иммуногистохимия; толуидиновый синий; сафранин; альциановый синий; триптаза; химаза.
2021, том 13, номер 4, стр. 93.
DOI: https://doi.org/10.17691/stm2021.13.4.10

Полный текст статьи

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Тучные клетки играют важную роль в защите организма от аллергенов, патогенов и паразитов, участвуя в развитии воспаления. В то же время имеются данные об их причастности к патогенезу ряда атопических, аутоиммунных, а также сердечно-сосудистых, онкологических, нервных и других заболеваний (аллергии, астмы, экземы, ринита, анафилаксии, мастоцитоза, рассеянного склероза, ревматоидного артрита, воспалительных заболеваний желудочно-кишечного тракта и легких, мигрени и т.д.). Диагностика многих заболеваний и изучение функций тучных клеток в норме и патологии требуют их выявления, вследствие чего особую важность приобретает знание о методах адекватной визуализации этих клеток у разных видов животных и человека.

В настоящем обзоре представлены данные об основных методах визуализации тучных клеток: ферментно-гистохимическом, иммуногистохимическом, а также гистохимии с использованием гистологических красителей. Основные гистологические красители соединяются с гепарином и другими кислыми мукополисахаридами, содержащимися в тучных клетках, и окрашивают их метахроматически. К их числу относятся толуидиновый синий, метиленовый синий (в том числе в составе краски Май-Грюнвальда–Гимзы), тионин, пинацианол и др. Сафранин и флуоресцентные красители берберин и авидин также соединяются с гепарином. Для маркирования мукозных и незрелых тучных клеток необходимо более продолжительное окрашивание гистологическими красителями или окраска альциановым синим.

Более современные методы — ферментно-гистохимический и особенно иммуногистохимический — позволяют высокоизбирательно выявлять тучные клетки с помощью реакции на триптазу или химазу (специфичные для данных клеток протеазы). При иммуногистохимическом исследовании триптаз и химаз следует учитывать видовые различия распределения этих протеаз в тучных клетках человека и животных для адекватного их выявления. Иммуногистохимическая реакция на рецептор к иммуноглобулину Е (FcεRI) и рецептор c-kit не специфична для тучных клеток, но последняя важна для демонстрации их пролиферации при нормальном и злокачественном росте.

В обзоре обсуждается также правильная фиксация биологического материала, что имеет большое значение для гистохимического и иммуногистохимического выявления тучных клеток.

Отмечено, что в качестве новых технологических подходов для анализа различных популяций тучных клеток применяются флуоресцентные методы иммуногистохимии и многомаркерный анализ в сочетании с конфокальной микроскопией.

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