Влияние пептидного блокатора кальциевых каналов ω-hexatoxin-Hv1a на клеточную гибель при ишемии/реперфузии in vitro

Апоптоз и некроз при реперфузии после ишемии являются ключевыми механизмами на клеточном уровне, приводящими к повреждению. Развитию патологических состояний предшествует внутриклеточная перегрузка ионами кальция как на стадии ишемии, так и на стадии реперфузии. В связи с этим одна из стратегий, направленных на уменьшение повреждений при ишемии/реперфузии, связана с использованием блокаторов кальциевых каналов.

Цель исследования — изучение влияния пептидного токсина — блокатора кальциевого канала ω-hexatoxin-Hv1a — на разные типы клеточной гибели эпителиальных клеток при воспроизведении in vitro условий ишемии/реперфузии, характерных при трансплантации органа.

Материалы и методы. В работе использовали эпителиальную клеточную культуру CHO-K1. Оценивали изменение апоптоза, некроза, клеточного индекса и концентрации ионов кальция при моделировании процессов ишемии/реперфузии in vitro с добавлением токсина — блокатора кальциевого канала. Ишемического и реперфузионного повреждения достигали за счет депривации кислорода и питательных веществ с последующей реперфузией в полной питательной среде. Измерения проводили с использованием мультимодального планшетного ридера-флуориметра.

Результаты. При моделировании процессов ишемии/реперфузии зафиксировано повышение апоптоза, некроза и концентрации ионов кальция. При добавлении токсина в концентрации 50 нМ на стадии реперфузии отмечено снижение апоптоза и некроза, а также концентрации ионов кальция до физиологического уровня или уровня, близкого к физиологическому. В присутствии токсина клеточный индекс показал более быстрое восстановление.

Заключение. Данные эксперимента подтверждают гипотезу о благотворном влиянии пептидных блокаторов кальциевых каналов на состояние эпителиальных клеток при реперфузии после ишемии и могут рассматриваться для дальнейшего изучения как стратегия адаптации органа перед реперфузией.

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