Abstract
Background . Ulcerative colitis is a colon disease accompanied by a systemic inflammatory response with the development of oxidative stress, which leads to structural and functional changes in cell membranes. There are no data on the qualitative and quantitative changes in the protein components of the cytoplasmic membrane in ulcerative colitis under the influence of the antioxidant, membrane-stabilizing drug Mexidol. The article presents the data of our own research, the purpose of which is to study the nature of changes in protein components in the erythrocyte membrane in vitro when exposed to Mexidol (the active substance is oxymethylethyl peridine succinate) in patients with ulcerative colitis during an acute attack. Materials and methods . The study involved 51 patients with ulcerative colitis during the exacerbation period and 30 clinically healthy individuals comparable by sex and age. Ten major protein components of the erythrocyte membrane were evaluated. Results . As a result of the regression analysis, it was shown that in the group of clinically healthy individuals, almost all structural proteins are to varying degrees linked to proteins by glucose transporters and the enzyme glutathione-S-transferase and in patients with ulcerative colitis, the connection of protein components of membrane channels with spectrines is lost. Incubation of red blood cells of patients with ulcerative colitis in a solution of Mexidol in vitro did not affect the protein components of the erythrocyte cytoplasmic membrane. At the same time, in ulcerative colitis patients there was a restoration of spectrin connections with anion transport proteins and a glucose transporter, which were not detected in the initial state. Conclusion . The effect of Mexidol in vitro on the erythrocyte membranes of patients with ulcerative colitis was accompanied by the restoration of structural and functional protein bonds, which can be considered as a beneficial effect of this drug on the erythrocyte membrane under the conditions of the studied pathology.
Highlights
a colon disease accompanied by a systemic inflammatory response with the development
The article presents the data of our own research
the purpose of which is to study the nature of changes in protein components
Summary
Исследование выполнено с соблюдением этических принципов медицинских исследований с участием человека, изложенных в Хельсинкской декларации Всемирной медицинской ассоциации, согласно протоколу, одобренному комитетом по этике ФГБНУ «Иркутский научный центр хирургии и травматологии» (протокол заседания No 9 от 9.11.2012 г.). Для получения цитоплазматических мембран отмытые эритроциты у каждого исследуемого разрушали осмотическим шоком по методу Dodge [6] после их 30-минутной инкубации в физиологическом растворе и 0,05%-ном «Мексидоле». По окончании гемолиза «тени» эритроцитов осаждали центрифугированием при 20000 g. Все операции по выделению и очистке водорастворимой фракции белков проводили в холодной комнате (+5 °С). Концентрацию общего белка определяли с использованием набора Qubit Protein Assay Kit (Invitrogen, США) на приборе Qubit согласно инструкции фирмы изготовителя. Электрофорез проводили по методу Лэммли [7] в полиакриламидном геле (ПААГ) с концентрацией разделяющего геля 7,5 и 15 % в присутствии SDS с использованием аппаратуры и реактивов фирмы Bio-Rad. На дорожки наносили по 10 мкг суммарного белка. Для определения вида белка на электрофореграмме набор маркеров фирмы Thermoscientific (#26614). Для оценки характера взаимосвязи между переменными применяли метод нелинейного регрессионного анализа
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