vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/VJ18.379vavilov-1546Research ArticleМОЛЕКУЛЯРНАЯ И КЛЕТОЧНАЯ БИОЛОГИЯMOLECULAR AND CELL BIOLOGYИССЛЕДОВАНИЕ ВЛИЯНИЯ ГИПОТЕРМИЧЕСКОЙ КОНСЕРВАЦИИ НА УРОВЕНЬ НАТРИЯ В КЛЕТКАХ ЭНДОТЕЛИЯ ТРАНСПЛАНТАТА РОГОВИЦЫSTUDY OF THE EFFECT OF HYPOTHERMIC CONSERVATION ON THE INTRACELLULAR SODIUM CONCENTRATION IN THE ENDOTHELIUM OF CORNEAL TRANSPLANTSБатуринаГ. С.BaturinaG. S.

Новосибирск

Novosibirsk

ПальчиковаИ. Г.PalchikovaI. G.

Новосибирск

Novosibirsk

КоневА. А.KonevA. A.

Новосибирск

Novosibirsk

СмирновЕ. С.SmirnovE. S.

Новосибирск

Novosibirsk

КатковаЛ. Е.KatkovaL. E.

Новосибирск

Novosibirsk

СоленовЕ. И.SolenovE. I.

Novosibirsk

eugsol@bionet.nsc.ruИскаковИ. А.IskakovI. А.Федеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наукРоссияInstitute of Cytology and Genetics SB RASRussian FederationКонструкторско-технологический институт научного приборостроения Сибирского отделения Российской академии наукРоссияTechnological Design Institute of Scientific Instrument Engineering SB RASRussian FederationФедеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наук; Новосибирский национальный исследовательский государственный университет; Новосибирский государственный технический университетРоссияInstitute of Cytology and Genetics SB RAS; Novosibirsk State University; Novosibirsk State Technical UniversityRussian FederationМежотраслевой научно-технический комплекс «Микрохирургия глаза» им. академика С.Н. Федорова Минздрава России, Новосибирский филиалРоссияS. Fyodorov Eye Microsurgery Federal State Institution, Novosibirsk BranchRussian Federation201804072018224433437Copyright © Батурина Г.С., Пальчикова И.Г., Конев А.А., Смирнов Е.С., Каткова Л.Е., Соленов Е.И., Искаков И.А., 20182018Батурина Г.С., Пальчикова И.Г., Конев А.А., Смирнов Е.С., Каткова Л.Е., Соленов Е.И., Искаков И.А.Baturina G.S., Palchikova I.G., Konev A.A., Smirnov E.S., Katkova L.E., Solenov E.I., Iskakov I.А.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/1546

Транспорт воды и ионов клетками эндотелия роговицы определяет ее жизнеспособность и оптические свойства. Исследовали влияние гипотермической консервации роговицы глаза на концентрацию натрия в клетках эндотелия роговицы. С этой целью определяли внутриклеточную кон центрацию натрия в клетках эндотелия роговицы глаза свиньи после гипотермической консервации при 4 °С в течение 1 и 10 суток и трансплантатов роговицы человека после 10 суток консервации. Концентрацию внутриклеточного натрия определяли флуориметрическим методом с помощью флуоресцентного красителя SodiumGreen в препаратах клеток эндотелия. Анализ флуоресцентных изображений клеток проводили с применением оригинальной программы CytoDynamics. Расчет концентраций натрия в клетках эндотелия роговицы свиньи выявил значительное повышение уровня внутриклеточного натрия после гипотермической консервации. Показано статистически значимое снижение проницаемости для натрия плазматических мембран клеток эндотелия после консервации. Уровень внутриклеточного натрия в клетках эндотелия препаратов роговицы человека после гипотермической консервации был выше, чем в аналогичных образцах эндотелия роговицы свиньи. Концентрация внутриклеточного натрия – перспективный интегральный показатель функциональной компетентности клеток эндотелия исследуемого образца роговицы.

Endothelial keratoplasty has become the treatment of choice for corneal endothelial dysfunction. Advancements in the surgical treatment of corneal endothelial diseases depend on progress in graft conservation and its related advantages in assessing the suitability of grafts for transplantation. Transport of water and ions by cornea endothelium is important for the optic properties of cornea. In this work, we study the intracellular sodium concentration in cornea endothelial cells in samples of pig cornea that underwent hypothermic conservation for 1 and 10 days and endothelial cells of human cornea grafts after 10-day conservation. The concentration of intracellular sodium in preparations of endothelial cells was assayed using fluorescent dye SodiumGreen. The fluorescent images were analyzed with the custom-made computer program CytoDynamics. An increased level of intracellular sodium was shown in the endothelium after 10-day conservation in comparison with one-day conservation (pig samples). Sodium permeability of pig endothelial cell plasma membranes significantly decreased in these samples. Assessment of intracellular sodium in human cornea endothelium showed a higher level – as was in analogues pig samples of the corneal endothelium. The assay of the intracellular sodium balance concentration established in endothelial cells after hypothermic conservation in mediums L-15 and Optisol-GS showed a significant advantage of specialized me dium Optisol-GS. The balanced intracellular concentration after 10 days of hypothermic conservation was significantly lower in cells incubated at 4 °C in Optisol-GS (L-15, 128 ± 14,  n = 15; Optisol-GS, 108 ± 14, n = 11; mM, p < 0.001). Intracellular sodium concentration could be a useful parameter for assessing cornea endothelium cell viability.

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