Russian Journal of Biological Physics and Chemisrty Russian Journal of Biological Physics and Chemisrty АКТУАЛЬНЫЕ ВОПРОСЫ БИОЛОГИЧЕСКОЙ ФИЗИКИ И ХИМИИ 2499-9962 54255 МЕДИЦИНСКАЯ БИОФИЗИКА И БИОФИЗИЧЕСКАЯ ХИМИЯ MEDICAL BIOPHYSICS AND BIOPHYSICAL CHEMISTRY МЕДИЦИНСКАЯ БИОФИЗИКА И БИОФИЗИЧЕСКАЯ ХИМИЯ MECHANISMS OF THE OXIDATIVE STRESS DEVELOPMENT UNDER ZINC IONS DEFICIENT STATE MODELING IN HUMAN ERYTHROCYTES IN VITRO МЕХАНИЗМЫ РАЗВИТИЯ ОКИСЛИТЕЛЬНОГО СТРЕССА ПРИ МОДЕЛИРОВАНИИ СОСТОЯНИЯ ДЕФИЦИТА ИОНОВ ЦИНКАВ ЭРИТРОЦИТАХ ЧЕЛОВЕКА INVITRO Гармаза Ю М Harmaza Y M garmaza@yandex.ru Тамашевский А В Tamashevski A V Институт биофизики и клеточной инженерии НАН Беларуси ru Institute of biophysics and cell engineering of National academy of sciences of Belarus ru Институт биофизики и клеточной инженерии НАН Беларуси ru Institute of biophysics and cell engineering of National academy of sciences of Belarus ru 25 03 2018 25 03 2018 3 1 115 122 20 03 2018 20 03 2018 https://rusjbpc.ru/en/nauka/article/54255/view

Установлено, что инкубация эритроцитов человека с внутриклеточным хелатором - N’,N’-тетракис-(2-пиридил-метил)-этилендиамином (TPEN), в субгемолитических концентрациях приводит к достоверному снижению внутриклеточного пула Zn2+ и увеличению эстеразной активности клеток. Продемонстрировано, что одним из возможных механизмов развития окислительного стресса в эритроцитах человека в условиях дефицита цинка является ингибирование активности основных ферментов антиоксидантной защиты - каталазы и глутатионпероксидазы и изменение концентрации восстановленного глутатиона. Выявлено, что именно ингибирование фермента глутатионпероксидаза вносит вклад в активацию эритроцитарных эстераз в условиях дефицита ионов цинка. Более того, обнаруженное усиление экспрессии цистеин-обогащенных белков металлотионеинов в эритроцитах человека при моделировании Zn-дефицитного состояния in vitro подтверждает предположение о функционировании данных белков в качестве вспомогательного антиоксиданта в защитной системе клеток.

It was established that incubation of human erythrocytes with an intracellular chelator N’,N’-tetrakis(2-methyl-pyridyl)ethylenediamine (TPEN) in subhemolytic concentrations leads to a significant decrease in the Zn2+ intracellular pool and cells esterase activity rise. It was demonstrated that one of the possible mechanisms of an oxidative stress development in zinc deficient human erythrocytes is the inhibition of the main antioxidant enzymes activity - catalase and glutathione peroxidase as well as changes in the reduced glutathione concentration. It was found that decreased glutathione peroxidase activity exactly contributes to the erythrocyte esterases activation under zinc ions deficiency. An increased expression of the cysteine-rich proteins metallothioneins in human erythrocytes under simulation of the Zn-deficient state in vitro was revealed. It confirms the hypothesis about the functioning of these proteins as an auxiliary antioxidant in a protective cell system.

 эритроциты человека дефицит цинка редокс-статус лабильный пул цинка эстеразная активность антиоксидантная система металлотионеины human erythrocytes zinc deficiency redox-state zinc labile pool esterase activity antioxidant system metallothioneins

Maret W., Li Y. Coordination dynamics of zinc in proteins. Chem. Rev., 2009, vol. 109, pp. 4682-4707. Maret W., Li Y. Coordination dynamics of zinc in proteins. Chem. Rev., 2009, vol. 109, pp. 4682-4707. Andreini C., Banci L., Bertini I., Rosato A. Counting the zinc-proteins encoded in the human genome. J. Proteome. Res., 2006, vol. 5, no. 1, pp. 196-201. Andreini C., Banci L., Bertini I., Rosato A. Counting the zinc-proteins encoded in the human genome. J. Proteome. Res., 2006, vol. 5, no. 1, pp. 196-201. Гармаза Ю.М., Слобожанина Е.И. Эccенциальноcть и токcичноcть цинка. Биофизичеcкие аcпекты. Биофизика, 2014, т. 59, вып. 2, с. 322-337. [Garmaza Yu.M., Slobozhanina E.I. Zinc essentiality and toxicity. Biophysical aspects. Biophysics, vol. 59, iss. 2, pp. 322-337. (In Russ.)] Garmaza Yu.M., Slobozhanina E.I. Eccencial'noct' i tokcichnoct' cinka. Biofizicheckie acpekty. Biofizika, 2014, t. 59, vyp. 2, s. 322-337. [Garmaza Yu.M., Slobozhanina E.I. Zinc essentiality and toxicity. Biophysical aspects. Biophysics, vol. 59, iss. 2, pp. 322-337. (In Russ.)] Prasad A.S., Halsted J.A., Nadimi M. Syndrome of iron deficiency anemia, hepatosplenomegaly, hypogonadism, dwarfism and geophagia. Am. J. Med., 1961. vol. 31, pp. 532-546. Prasad A.S., Halsted J.A., Nadimi M. Syndrome of iron deficiency anemia, hepatosplenomegaly, hypogonadism, dwarfism and geophagia. Am. J. Med., 1961. vol. 31, pp. 532-546. Moynahan E.J. Letter: Acrodermatitis enteropathica: A lethal inherited human zinc-deficiency disorder. Lancet, 1974, vol. 2, pp. 399-400. Moynahan E.J. Letter: Acrodermatitis enteropathica: A lethal inherited human zinc-deficiency disorder. Lancet, 1974, vol. 2, pp. 399-400. Wang K., Zhou B., Kuo Y.M., Zemansky J., Gitschier J. A novel member of a zinc transporter family is defective in acrodermatitis enteropathica. Am. J. Hum. Genet., 2002, vol. 71, pp. 66-73. Wang K., Zhou B., Kuo Y.M., Zemansky J., Gitschier J. A novel member of a zinc transporter family is defective in acrodermatitis enteropathica. Am. J. Hum. Genet., 2002, vol. 71, pp. 66-73. Prasad A.S. Impact of the discovery of human zinc deficiency on health. J. Trace Elem. Med. Biol., 2014, vol. 28, no. 4, pp. 357-363. Prasad A.S. Impact of the discovery of human zinc deficiency on health. J. Trace Elem. Med. Biol., 2014, vol. 28, no. 4, pp. 357-363. Zago M.P., Oteiza P.I. The antioxidant properties of zinc: interactions with iron and antioxidants. Free Radic. Biol. Med., 2001, vol. 31, no. 2, pp. 266-274. Zago M.P., Oteiza P.I. The antioxidant properties of zinc: interactions with iron and antioxidants. Free Radic. Biol. Med., 2001, vol. 31, no. 2, pp. 266-274. Oteiza P.I., Clegg M.S., Zago M.P., Keen C.L. Zinc deficiency induces oxidative stress and AP-1 activation in 3T3 cells. Free Radical Biol. Med., 2000, vol. 28, pp. 1091-1099. Oteiza P.I., Clegg M.S., Zago M.P., Keen C.L. Zinc deficiency induces oxidative stress and AP-1 activation in 3T3 cells. Free Radical Biol. Med., 2000, vol. 28, pp. 1091-1099. Ho E., Ames B.N. Low intracellular zinc induces oxidative DNA damage, disrupts p53, Nfkappa B, and AP1 DNA binding, and affects DNA repair in a rat glioma cell line. Proc. Natl. Acad. Sci. U.S.A., 2002, vol. 99, pp. 16770-16775. Ho E., Ames B.N. Low intracellular zinc induces oxidative DNA damage, disrupts p53, Nfkappa B, and AP1 DNA binding, and affects DNA repair in a rat glioma cell line. Proc. Natl. Acad. Sci. U.S.A., 2002, vol. 99, pp. 16770-16775. Song Y., Leonard S.W., Traber M.G., Ho E. Zinc deficiency affects DNA damage, oxidative stress, antioxidant defenses, and DNA repair in rats. J. Nutr., 2009, vol. 139, pp. 1626-1631. Song Y., Leonard S.W., Traber M.G., Ho E. Zinc deficiency affects DNA damage, oxidative stress, antioxidant defenses, and DNA repair in rats. J. Nutr., 2009, vol. 139, pp. 1626-1631. Brennan A.M., Suh S.W., Won S.J., Narasimhan P., Kauppinen T.M., Lee H., Edling Y., Chan P.H., Swanson R.A. NADPH oxidase is the primary source of superoxide induced by NMDA receptor activation. Nat. Neurosci., 2009, vol. 12, pp. 857-863. Brennan A.M., Suh S.W., Won S.J., Narasimhan P., Kauppinen T.M., Lee H., Edling Y., Chan P.H., Swanson R.A. NADPH oxidase is the primary source of superoxide induced by NMDA receptor activation. Nat. Neurosci., 2009, vol. 12, pp. 857-863. Paoletti A.M., Vergnano A.M., Barbour B., Casado M. Zinc at glutamatergic synapses. Neuroscience, 2009, vol. 158, pp. 126-136. Paoletti A.M., Vergnano A.M., Barbour B., Casado M. Zinc at glutamatergic synapses. Neuroscience, 2009, vol. 158, pp. 126-136. Gee K.R., Zhou Z.L., Ton-That D., Sensi S.L., Weiss J.H. Measuring zinc in living cells. A new generation of sensitive and selective fluorescent probes. Cell Calcium, 2002, vol. 31, no. 5, pp. 245-251. Gee K.R., Zhou Z.L., Ton-That D., Sensi S.L., Weiss J.H. Measuring zinc in living cells. A new generation of sensitive and selective fluorescent probes. Cell Calcium, 2002, vol. 31, no. 5, pp. 245-251. Bratosin D., Mitrofan L., Palli C., Estaquier J. Novel fluorescence assay using Calcein-AM for the determination of human erythrocyte viability and aging. Cytometry A, 2005, vol. 66A, pp. 78-84. Bratosin D., Mitrofan L., Palli C., Estaquier J. Novel fluorescence assay using Calcein-AM for the determination of human erythrocyte viability and aging. Cytometry A, 2005, vol. 66A, pp. 78-84. Ellman G.L. Tissue sulfhydryl groups. Arch Biochem Biophys., 1959, vol. 82, no. 1, pp. 70-77. Ellman G.L. Tissue sulfhydryl groups. Arch Biochem Biophys., 1959, vol. 82, no. 1, pp. 70-77. Моин В.М. Простой и специфический метод определения активности глутатионпероксидазы в эритроцитах. Лаб. Дело, 1986, № 12, с. 724-727. [Moin V.M. A simple and specific method for determining of the glutathione peroxidase activity in erythrocytes. Laboratory diagnostics, 1986, no. 12, pp. 724-727. (In Russ)] Moin V.M. Prostoy i specificheskiy metod opredeleniya aktivnosti glutationperoksidazy v eritrocitah. Lab. Delo, 1986, № 12, s. 724-727. [Moin V.M. A simple and specific method for determining of the glutathione peroxidase activity in erythrocytes. Laboratory diagnostics, 1986, no. 12, pp. 724-727. (In Russ)] Королюк М.А., Иванова Л.И., Майорова И.Г., Токарева В.Е. Метод определения каталазной активности. Лаб. дело, 1988. № 1. c. 16-19. [Koroluk М.А., Ivanova L.I., Majorova I.G., Tokareva V.Е. Method for the determination of catalase activity [Metod opredeleniya katalaznoy aktivnosti]. Laboratory diagnostics, 1988, no. 1, pp. 16-19. (in Russ)] Korolyuk M.A., Ivanova L.I., Mayorova I.G., Tokareva V.E. Metod opredeleniya katalaznoy aktivnosti. Lab. delo, 1988. № 1. c. 16-19. [Koroluk M.A., Ivanova L.I., Majorova I.G., Tokareva V.E. Method for the determination of catalase activity [Metod opredeleniya katalaznoy aktivnosti]. Laboratory diagnostics, 1988, no. 1, pp. 16-19. (in Russ)] Anti-Metallothionein antibody [UC1MT]. 04.10.2015, URL: abcam.com/metallothionein-antibody-uc1mt-ab12228.html. Anti-Metallothionein antibody [UC1MT]. 04.10.2015, URL: abcam.com/metallothionein-antibody-uc1mt-ab12228.html. McCabe M.J., Jiang S.A., Orrenius S. Chelation of intracellular zinc triggers apoptosis in mature thymocytes. Lab. Invest., 1993, vol. 69, pp. 101-110. McCabe M.J., Jiang S.A., Orrenius S. Chelation of intracellular zinc triggers apoptosis in mature thymocytes. Lab. Invest., 1993, vol. 69, pp. 101-110. Harmaza Y., Slobozhanina E. Zinc homeostasis and eryptosis. FEBS J., 2013, vol. 280, suppl. 1, p. 218. Harmaza Y., Slobozhanina E. Zinc homeostasis and eryptosis. FEBS J., 2013, vol. 280, suppl. 1, p. 218. Гармаза Ю.М., Тамашевский А.В., Гончарова Н.В., Слобожанина Е.И. Влияние внутриклеточного уровня цинка в эритроцитах человека на перераспределение фосфатидилсерина и их жизнеспособность. Новости медико-биологических наук, 2011, т. 3, № 1, с. 90-95. [Harmaza Y.M., Tamashevski A.V., Goncharova N.V., Slobozhanina E.I. News of biomedical sciences, 2011, vol, 3, no. 1, pp. 90-95. (in Russ)] Garmaza Yu.M., Tamashevskiy A.V., Goncharova N.V., Slobozhanina E.I. Vliyanie vnutrikletochnogo urovnya cinka v eritrocitah cheloveka na pereraspredelenie fosfatidilserina i ih zhiznesposobnost'. Novosti mediko-biologicheskih nauk, 2011, t. 3, № 1, s. 90-95. [Harmaza Y.M., Tamashevski A.V., Goncharova N.V., Slobozhanina E.I. News of biomedical sciences, 2011, vol, 3, no. 1, pp. 90-95. (in Russ)] Гармаза Ю.М., Тамашевский А.В., Канаш Ю.С., Зубрицкая Г.П., Кутько А.Г., Слобожанина Е.И. Внутриклеточный цинк: роль в Н2О2-индуцированном окислительном стрессе в эритроцитах человека. Биофизика, 2016, т. 61, вып. 6, с. 1149-1158. [Harmaza Y.M., Tamashevski A.V., Kanash J.S., Zubritskaya G.P., Kutko A.G., Slobozhanina E.I. Intracellular Zinc: a Role in H2O2-Induced Oxidative Stress in Human Erythrocytes. Biophysics, 2016, vol. 61, iss. 6, pp. 1149-1158. (in Russ)] Garmaza Yu.M., Tamashevskiy A.V., Kanash Yu.S., Zubrickaya G.P., Kut'ko A.G., Slobozhanina E.I. Vnutrikletochnyy cink: rol' v N2O2-inducirovannom okislitel'nom stresse v eritrocitah cheloveka. Biofizika, 2016, t. 61, vyp. 6, s. 1149-1158. [Harmaza Y.M., Tamashevski A.V., Kanash J.S., Zubritskaya G.P., Kutko A.G., Slobozhanina E.I. Intracellular Zinc: a Role in H2O2-Induced Oxidative Stress in Human Erythrocytes. Biophysics, 2016, vol. 61, iss. 6, pp. 1149-1158. (in Russ)] Гармаза Ю.М., Тамашевский А.В., Слобожанина Е.И. Металлотионеины млекопитающих: структура и биологическая роль. Известия НАН Беларуси. Серия биол. наук, 2016, no. 1, с. 107-116. [Harmaza Y.M., Tamashevski A.V., Slobozhanina E.I. Mammalian metalothioneins: structure and biological role. Proceedings of the National academy of sciences of Belarus, 2016, no. 1, pp. 107-116. (in Russ.)] Garmaza Yu.M., Tamashevskiy A.V., Slobozhanina E.I. Metallotioneiny mlekopitayuschih: struktura i biologicheskaya rol'. Izvestiya NAN Belarusi. Seriya biol. nauk, 2016, no. 1, s. 107-116. [Harmaza Y.M., Tamashevski A.V., Slobozhanina E.I. Mammalian metalothioneins: structure and biological role. Proceedings of the National academy of sciences of Belarus, 2016, no. 1, pp. 107-116. (in Russ.)] Min K.S., Tanaka N., Horie T., Kawano H., Tetsuchikawahara N., Onosaka S. Metalothionein-enriched hepatocytes are resistant to ferric nitriloacetate toxicity during conditions of glutathione depletion. Toxicol. Lett., 2005, vol. 158, pp. 108-115. Min K.S., Tanaka N., Horie T., Kawano H., Tetsuchikawahara N., Onosaka S. Metalothionein-enriched hepatocytes are resistant to ferric nitriloacetate toxicity during conditions of glutathione depletion. Toxicol. Lett., 2005, vol. 158, pp. 108-115. Gumulec J., Masarik M., Krizkova S., Adam V., Hubalek J., Hrabeta J., Eckschlager T., Stiborova M., Kizek R. Insight to physiology and pathology of zinc(II) ions and their actions in breast and prostate carcinoma. Curr. Med. Chem., 2011, vol. 18, pp. 5041-5051. Gumulec J., Masarik M., Krizkova S., Adam V., Hubalek J., Hrabeta J., Eckschlager T., Stiborova M., Kizek R. Insight to physiology and pathology of zinc(II) ions and their actions in breast and prostate carcinoma. Curr. Med. Chem., 2011, vol. 18, pp. 5041-5051. Formigari A., Santon A., Irato P. Efficacy of zinc treatment against iron-induced toxicity in rat hepatoma cell line H4-II-E-C3. Liver Int., 2007, vol. 27, pp. 120-127. Formigari A., Santon A., Irato P. Efficacy of zinc treatment against iron-induced toxicity in rat hepatoma cell line H4-II-E-C3. Liver Int., 2007, vol. 27, pp. 120-127. Eide D.J. The oxidative stress of zinc deficiency. Metallomics, 2011, vol. 3. pp. 1124-1129. Eide D.J. The oxidative stress of zinc deficiency. Metallomics, 2011, vol. 3. pp. 1124-1129.