Современные методы исследования апоптотической гибели клеток (обзор)

Рассмотрены современные методы изучения апоптотической гибели клеток in vitro и in vivo. Показано, что наблюдение апоптоза основано на регистрации характерных клеточных изменений: потери целостности и деполяризации плазматической мембраны, экспозиции фосфатидилсерина и фосфатидилэтаноламина на поверхности клеточной мембраны, активации каспаз, деполяризации митохондриальной мембраны, фрагментации ДНК, изменении уровня биохимических маркеров. Основными методами исследования апоптоза in vitro являются: световая и электронная микроскопия, проточная цитометрия, флюоресцентная микроскопия, иммуногистохимия, иммуноферментный анализ, вестерн-блот, электрофорез и TUNEL-метод. Важную роль в наше время играют неинвазивные методы изучения апоптоза in vivo с возможностью регистрации этого процесса в тканях, органах и организме в целом: позитронно-эмиссионная томография (ПЭТ), однофотонная эмиссионная компьютерная томография (ОФЭКТ), магнитно-резонансная томография (МРТ), флюоресцентная спектроскопия, FLIM/FRET-имиджинг. Большое внимание уделено поиску контрастных агентов, специфичных к различным молекулам апоптотического каскада. Показано, что характер апоптотических реакций может служить маркером эффективности рациональной химиотерапии и помогать в ее выборе. Отмечено, что все бльшую популярность приобретает комбинация нескольких методов исследования апоптотической гибели клеток, например ОФЭКТ и оптический имиджинг, ПЭТ и оптический имиджинг, ПЭТ и МРТ. В перспективе с помощью этих методов можно будет не только определять тяжесть заболевания, но и оценивать эффективность лечения.

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