medlitГигиена и санитарияHygiene and Sanitation0016-99002412-0650Federal Scientific Center of Hygiene named after F.F. Erisman10.47470/0016-9900-2017-96-7-690-694medlit-876Research ArticleПРОФИЛАКТИЧЕСКАЯ ТОКСИКОЛОГИЯ И ГИГИЕНИЧЕСКОЕ НОРМИРОВАНИЕPREVENTIVE TOXICOLOGY AND HYGIENIC STANDARTIZATIONСоотношение процессов клеточной пролиферации и апоптоза в коже при воздействии солей тяжелых металлов и хелаторов эссенциальных металловRatio of processes of cell proliferation and apoptosis in the skin under exposure to heavy metal salts and chelators of essential metalshttps://orcid.org/0000-0003-2514-6332НиколаеваТатьяна ВладимировнаNikolaevaTatyana V.

Кандидат медицинских наук, доцент кафедры дерматовенерологии ГБОУ ВПО «Оренбургский государственный медицинский университет» Минздрава России, 460000, Оренбург.

e-mail: orenderma@yandex.ru

MD, PhD, аssociate Professor of the Department of dermatology of the Orenburg State Medical UniversityUniversity, Orenburg, 460000, Russian Federation.

e-mail: orenderma@yandex.ru

orenderma@yandex.ruhttps://orcid.org/0000-0003-4100-3630ПоляковаВ. С.PolyakovaV. S.noemail@neicon.ruhttps://orcid.org/0000-0001-6698-2164СеткоН. П.SetkoN. P.noemail@neicon.ruhttps://orcid.org/0000-0003-4834-8324ВоронинаЛ. Г.VoroninaL. G.noemail@neicon.ruГБОУ ВПО «Оренбургский государственный медицинский университет» Министерства здравоохранения РоссииOrenburg State Medical University201721102020967690694Copyright © Николаева Т.В., Полякова В.С., Сетко Н.П., Воронина Л.Г., 20202020Николаева Т.В., Полякова В.С., Сетко Н.П., Воронина Л.Г.Nikolaeva T.V., Polyakova V.S., Setko N.P., Voronina L.G.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.rjhas.ru/jour/article/view/876

В модельном эксперименте на мышах линии C57BL/6 установлены особенности влияния тяжелых металлов и хелаторов эссенциальных металлов на пролиферацию и апоптоз эпителиальных клеток кожи (кератиноцитов). В рамках исследования 40 экспериментальных животных были распределены в семь опытных и контрольную группу по пять особей в каждой. Пролиферативную и апоптотическую активность кератиноцитов определяли иммуногистохимическим методом и оценивали путем расчета индекса пролиферации и индекса апоптоза в клетках покровного эпителия и эпителиальных клетках волосяных фолликулов в стадию, соответствовавшую позднему анагену. Введение животным сульфата цинка, бихромата натрия и хелатора цинка (N,N,N`,N`-тетракис(2-пиридилметил)-этилендиамин — ТПЭН) приводило к статистически значимому увеличению пролиферативной активности кератиноцитов. Снижение индекса пролиферации выявлено у животных, получавших ацетат свинца и хелатор меди (тетратиомолибдат аммония — ТТМ). Введение сульфата никеля и хелатора железа (дефероксамина) не повлияло на пролиферативную активность кератиноцитов. Индукция апоптоза эпителиальных клеток отмечена при поступлении в организм животных сульфата никеля, бихромата натрия, ацетата свинца, хелатора меди (ТТМ) и хелатора цинка (ТПЭН). У мышей, получавших сульфат цинка и дефероксамин, апоптотическая активность кератиноцитов не изменилась. Применение кластерного анализа позволило классифицировать вводившиеся экспериментальным животным вещества с учетом их одновременного влияния на исследуемые клеточные процессы. Показано, что ацетат свинца, хелатор железа (дефероксамин) и хелатор меди (тетратиомолибдат аммония) снижают пролиферативную активность кератиноцитов и незначительно влияют на апоптоз эпителиальных клеток кожи. Сульфат цинка, сульфат никеля, бихромат натрия и хелатор цинка (ТПЭН) активируют клеточную пролиферацию и индуцируют апоптоз кератиноцитов.

In the model experiment on C57BL /6 mice there were established features of the impact of heavy metals and chelators of essential metals on proliferation and apoptosis of epithelial skin cells (keratinocytes). For the execution of a study 40 test animals were divided into seven experimental and 1 control groups, each consisted of five animals. The proliferative and apoptotic activity of keratinocytes was determined by the immunohistochemical method and evaluated by calculating the proliferation index and the index of apoptosis in the cells of the surface epithelium and the epithelial cells of hair follicles in the late anagen stage. Comparative analysis of the proliferation index of the control group and experimental groups showed administration of zinc sulfate, sodium dichromate and zinc chelator (N, N, N`, N`-tetrakis (2-pyridylmethyl) ethylenediamine) to animals to give rise in a statistically significant increase in the proliferative activity of keratinocytes. The decline of proliferation index was detected in animals treated with lead acetate and copper chelator (ammonium tetrathiomolybdate). Introduction of an iron chelator (deferoxamine) had no effect on the proliferative activity of keratinocytes in experimental animals. Induction of apoptosis of epithelial cell was noted under the administration of nickel sulfate, sodium dichromate, lead acetate and zinc chelator (N, N, N`, N`-tetrakis (2-pyridylmethyl) ethylenediamine) to animals. In mice received deferoxamine zinc sulfate and apoptotic activity of keratinocytes has not changed. The use of cluster analysis allowed to classify substances administered to experimental animals, taking into account their simultaneous effect on the studied cellular processes. Lead acetate, iron chelator (deferoxamine) and copper chelator (ammonium tetrathiomolybdate) were shown to reduce the proliferative activity of keratinocytes and have little effect on apoptosis of the epithelial cells of the skin. Zinc sulfate, nickel sulfate, sodium dichromate and zinc chelator (N, N, N`, N`-tetrakis (2-pyridylmethyl) ethylenediamine) activate cell proliferation and induce apoptosis of keratinocytes.

тяжелые металлыхелаторы ессенциальных металловкожакератиноцитыиндекс пролиферациииндекс апоптозаheavy metalschelators of essentsial metalsskinkeratinocytesproliferation indexapoptosis indexReferencesКлишов А.А. Гистогенез и регенерация тканей. Ленинград: Медицина; 1984.Klishov A.A. Histogenesis and Tissue Regeneration [Gistogenez i regeneratsiya tkaney]. Leningrad: Meditsina; 1984. (in Russian)Ярилин А.А. Апоптоз. Природа феномена и его роль в целостном организме. Патологическая физиология и экспериментальная терапия. 1998; (2): 38-48.Yarilin A.A. Apoptosis. The nature of the phenomenon and its role in the whole organism. Patologicheskaya fiziologiya i eksperimental’naya terapiya. 1998; (2): 38–48. (in Russian)Peled T., Landau E., Prus E., Treves A.J., Nagler A., Fibach E. Cellular copper content modulates differentiation and self-renewal in cultures of cord blood-derived CD34+ cells. Br. J. Haematol. 2002; 116(3): 655-61.Peled T., Landau E., Prus E., Treves A.J., Nagler A., Fibach E. Cellular copper content modulates differentiation and self-renewal in cultures of cord blood-derived CD34+ cells. Br. J. Haematol. 2002; 116(3): 655–61.García-Rodríguez Mdel C., Carvente-Juárez M.M., Altamirano-Lozano M.A. Antigenotoxic and apoptotic activity of green tea polyphenol extracts on hexavalent chromium-induced DNA damage in peripheral blood of CD-1 mice: analysis with differential acridine orange/ethidium bromide staining. Oxid. Med. Cell Longev. 2013; 2013: 486419.García-Rodríguez Mdel C., Carvente-Juárez M.M., Altamirano-Lozano M.A. Antigenotoxic and apoptotic activity of green tea polyphenol extracts on hexavalent chromium-induced DNA damage in peripheral blood of CD-1 mice: analysis with differential acridine orange/ethidium bromide staining. Oxid. Med. Cell Longev. 2013; 2013: 486419.Su L., Deng Y., Zhang Y., Li C., Zhang R., Sun Y., et al. Protective effects of grape seed procyanidin extract against nickel sulfate-induced apoptosis and oxidative stress in rat testes. Toxicol. Mech. Methods. 2011; 21(6): 487-94.Su L., Deng Y., Zhang Y., Li C., Zhang R., Sun Y., et al. Protective effects of grape seed procyanidin extract against nickel sulfate-induced apoptosis and oxidative stress in rat testes. Toxicol. Mech. Methods. 2011; 21(6): 487–94.Tang K., Guo H., Deng J., Cui H., Peng X., Fang J., et al. Inhibitive effects of nickel chloride (NiCl2) on thymocytes. Biol. Trace Elem. Res. 2015; 164(2): 242-2.Tang K., Guo H., Deng J., Cui H., Peng X., Fang J., et al. Inhibitive effects of nickel chloride (NiCl2) on thymocytes. Biol. Trace Elem. Res. 2015; 164(2): 242–2.Quan F.S., Yu X.F., Gao Y., Ren W.Z. Protective effects of folic acid against central nervous system neurotoxicity induced by lead exposure in rat pups. Genet. Mol. Res. 2015; 14(4): 12466-71.Quan F.S., Yu X.F., Gao Y., Ren W.Z. Protective effects of folic acid against central nervous system neurotoxicity induced by lead exposure in rat pups. Genet. Mol. Res. 2015; 14(4): 12466–71.Müller-Röver S., Handjiski B., van der Veen C., Eichmüller S., Foitzik K., McKay I.A., et al. A comprehensive guide for the accurate classification of murine hair follicles in distinct hair cycle stages. J. Invest. Dermatol. 2001; 117(1): 3-15.Müller-Röver S., Handjiski B., van der Veen C., Eichmüller S., Foitzik K., McKay I.A., et al. A comprehensive guide for the accurate classification of murine hair follicles in distinct hair cycle stages. J. Invest. Dermatol. 2001; 117(1): 3–15.Paus R., Handjiski B., Eichmüller S., Czarnetzki B.M. Chemotherapy-induced alopecia in mice. Induction by cyclophosphamide, inhibition by cyclosporine A, and modulation by dexamethasone. Am. J. Pathol. 1994; 144(4): 719-34.Paus R., Handjiski B., Eichmüller S., Czarnetzki B.M. Chemotherapy-induced alopecia in mice. Induction by cyclophosphamide, inhibition by cyclosporine A, and modulation by dexamethasone. Am. J. Pathol. 1994; 144(4): 719–34.Zhang Y., Zhang Y., Xie Y., Gao Y., Ma J., Yuan J., et al. Multitargeted inhibition of hepatic fibrosis in chronic iron-overloaded mice by Salvia miltiorrhiza. J. Ethnopharmacol. 2013; 148(2): 671-81.Zhang Y., Zhang Y., Xie Y., Gao Y., Ma J., Yuan J., et al. Multitargeted inhibition of hepatic fibrosis in chronic iron-overloaded mice by Salvia miltiorrhiza. J. Ethnopharmacol. 2013; 148(2): 671–81.Wei H., Frei B., Beckman J.S., Zhang W.J. Copper chelation by tetrathiomolybdate inhibits lipopolysaccharide-induced inflammatory responses in vivo. Am. J. Physiol. Heart Circ. Physiol. 2011; 301(3): H712-20.Wei H., Frei B., Beckman J.S., Zhang W.J. Copper chelation by tetrathiomolybdate inhibits lipopolysaccharide-induced inflammatory responses in vivo. Am. J. Physiol. Heart Circ. Physiol. 2011; 301(3): H712–20.Fukuyama S., Matsunaga Y., Zhanghui W., Noda N., Asai Y., Moriwaki A., et al. A zinc chelator TPEN attenuates airway hyperresponsiveness and airway inflammation in mice in vivo. Allergol. Int. 2011; 60(3): 259-66.Fukuyama S., Matsunaga Y., Zhanghui W., Noda N., Asai Y., Moriwaki A., et al. A zinc chelator TPEN attenuates airway hyperresponsiveness and airway inflammation in mice in vivo. Allergol. Int. 2011; 60(3): 259–66.Бабиченко И.И., Ковязин В.А. Новые методы иммуногистохимической диагностики опухолевого роста: Учебное пособие. М.: РУДН; 2008Babichenko I.I., Kovyazin V.A. New Methods of Immunohistochemical Diagnostic of Tumor Growth: Textbook [Novye metody immunogistokhimicheskoy diagnostiki opukholevogo rosta: Uchebnoe posobie]. Moscow: RUDN; 2008. (in Russian)Earnshaw W.C., Martins L.M., Kaufmann S.H. Mammalian caspases: structure, activation, substrates and functions during apoptosis. Annu. Rev. Biochem. 1999; 68: 383-424Earnshaw W.C., Martins L.M., Kaufmann S.H. Mammalian caspases: structure, activation, substrates and functions during apoptosis. Annu. Rev. Biochem. 1999; 68: 383–424.Яровая Г.А., Нешкова Е.А., Мартынова Е.А., Блохина Т.Б. Роль протеолитических ферментов в контроле различных стадий апоптоза. Лабораторная медицина. 2011; (11): 39-53Yarovaya G.A., Neshkova E.A., Martynova E.A., Blokhina T.B. The role of the proteolytic enzyme in the control of different stages of apoptosis. Laboratornaya meditsina. 2011; (11): 39–53. (in Russian)Glantz S.A. Primer of Biostatistics. 5th edition. New York: McGraw-Hill; 2002Glantz S.A. Primer of Biostatistics. 5th edition. New York: McGraw-Hill; 2002.Айвазян С.А., Мхитарян С.А. Прикладная статистика. Основы эконометрики: учебник для вузов: в 2 т. Т. 1: Теория вероятностей и прикладная статистика. М.: ЮНИТИ-ДАНА; 2001.Ayvazyan S.A., Mkhitaryan S.A. Applied Statistics. Basics of Econometrics: Textbook for Institutes of Higher Education: in 2 Vol. Vol. 1: Probability Theory and Applied Statistics [Prikladnaya statistika. Osnovy ekonometriki: uchebnik dlya vuzov: v 2 t. T. 1: Teoriya veroyatnostey i prikladnaya statistika]. Moscow: YuNITI-DANA; 2001. (in Russian)Боев В.М., Борщук Е.Л., Екимов А.К., Бегун Д.Н. Руководство по обеспечению решения медико-биологических задач с применением программы Statistica 10.0. Оренбург: ИПК «Южный Урал»; 2014.Boev V.M., Borshchuk E.L., Ekimov A.K., Begun D.N. Guidelines for solving medical-biological problems using the program Statistica 10.0. Orenburg: IPK «Yuzhnyy Ural»; 2014. (in Russian)Kang X., Song Z., McClain C.J., Kang Y.J., Zhou Z. Zinc supplementation enhances hepatic regeneration by preserving hepatocyte nuclear factor-4alpha in mice subjected to long-term ethanol administration. Am. J. Pathol. 2008; 172(4): 916-25.Kang X., Song Z., McClain C.J., Kang Y.J., Zhou Z. Zinc supplementation enhances hepatic regeneration by preserving hepatocyte nuclear factor-4alpha in mice subjected to long-term ethanol administration. Am. J. Pathol. 2008; 172(4): 916–25.Azman M.S., Wan Saudi W.S., Ilhami M., Mutalib M.S., Rahman M.T. Zinc intake during pregnancy increases the proliferation at ventricular zone of the newborn brain. Nutr. Neurosci. 2009; 12(1): 9-12.Azman M.S., Wan Saudi W.S., Ilhami M., Mutalib M.S., Rahman M.T. Zinc intake during pregnancy increases the proliferation at ventricular zone of the newborn brain. Nutr. Neurosci. 2009; 12(1): 9–12.Kang M., Zhao L., Ren M., Deng M., Li C. Reduced metallothionein expression induced by Zinc deficiency results in apoptosis in hepatic stellate cell line LX-2. Int. J. Clin. Exp. Med. 2015; 8(11): 20603-9.Kang M., Zhao L., Ren M., Deng M., Li C. Reduced metallothionein expression induced by Zinc deficiency results in apoptosis in hepatic stellate cell line LX-2. Int. J. Clin. Exp. Med. 2015; 8(11): 20603–9.Shen H., Qin H., Guo J. Cooperation of metallothionein and zinc transporters for regulating zinc homeostasis in human intestinal Caco-2 cells. Nutr. Res. 2008; 28(6): 406-13.Shen H., Qin H., Guo J. Cooperation of metallothionein and zinc transporters for regulating zinc homeostasis in human intestinal Caco-2 cells. Nutr. Res. 2008; 28(6): 406–13.Hashemi M., Ghavami S., Eshraghi M., Booy E.P., Los M. Cytotoxic effects of intra and extracellular zinc chelation on human breast cancer cells. Eur. J. Pharmacol. 2007; 557(1): 9-19.Hashemi M., Ghavami S., Eshraghi M., Booy E.P., Los M. Cytotoxic effects of intra and extracellular zinc chelation on human breast cancer cells. Eur. J. Pharmacol. 2007; 557(1): 9–19.Itoh S., Kim H.W., Nakagawa O., Ozumi K., Lessner S.M., Aoki H., et al. Novel role of antioxidant-1 (Atox1) as a copper-dependent transcription factor involved in cell proliferation. J. Biol. Chem. 2008; 283(14): 9157-67.Itoh S., Kim H.W., Nakagawa O., Ozumi K., Lessner S.M., Aoki H., et al. Novel role of antioxidant-1 (Atox1) as a copper-dependent transcription factor involved in cell proliferation. J. Biol. Chem. 2008; 283(14): 9157–67.Beaver L.M., Stemmy E.J., Schwartz A.M., Damsker J.M., Constant S.L., Ceryak S.M., et al. Lung inflammation, injury, and proliferative response after repetitive particulate hexavalent chromium exposure. Environ. Health Perspect. 2009; 117(12): 1896-902.Beaver L.M., Stemmy E.J., Schwartz A.M., Damsker J.M., Constant S.L., Ceryak S.M., et al. Lung inflammation, injury, and proliferative response after repetitive particulate hexavalent chromium exposure. Environ. Health Perspect. 2009; 117(12): 1896–902.Sarkar A., Chattopadhyay S., Kaul R., Pal J.K. Lead exposure and heat shock inhibit cell proliferation in human HeLa and K562 cells by inducing expression and activity of the heme-regulated eIF-2alpha kinase. J. Biochem. Mol. Biol. Biophys. 2002; 6(6): 391-6.Sarkar A., Chattopadhyay S., Kaul R., Pal J.K. Lead exposure and heat shock inhibit cell proliferation in human HeLa and K562 cells by inducing expression and activity of the heme-regulated eIF-2alpha kinase. J. Biochem. Mol. Biol. Biophys. 2002; 6(6): 391–6.Zhang J., Cao H., Zhang Y., Zhang Y., Ma J., Wang J., et al. Nephroprotective effect of calcium channel blockers against toxicity of lead exposure in mice. Toxicol. Lett. 2013; 218(3): 273-80.Zhang J., Cao H., Zhang Y., Zhang Y., Ma J., Wang J., et al. Nephroprotective effect of calcium channel blockers against toxicity of lead exposure in mice. Toxicol. Lett. 2013; 218(3): 273–80.Rana S.V. Metals and apoptosis: recent developments. J. Trace Elem. Med. Biol. 2008; 22(4): 262-84.Rana S.V. Metals and apoptosis: recent developments. J. Trace Elem. Med. Biol. 2008; 22(4): 262–84.Bustos R.I., Jensen E.L., Ruiz L.M., Rivera S., Ruiz S., Simon F., et al. Copper deficiency alters cell bioenergetics and induces mitochondrial fusion through up-regulation of MFN2 and OPA1 in erythropoietic cells. Biochem. Biophys. Res. Commun. 2013; 437(3): 426-32.Bustos R.I., Jensen E.L., Ruiz L.M., Rivera S., Ruiz S., Simon F., et al. Copper deficiency alters cell bioenergetics and induces mitochondrial fusion through up-regulation of MFN2 and OPA1 in erythropoietic cells. Biochem. Biophys. Res. Commun. 2013; 437(3): 426–32.Jimenez-Cervantes C., Martinez-Esparza M., Perez C., Daum N., Solano F., Garcia-Borron J.C. Inhibition of melanogenesis in response to oxidative stress: transient downregulation of melanocyte differentiation markers and possible involvement of microphthalmia transcription factor. J. Cell Sci. 2001; 114(12): 2335-44.Jimenez-Cervantes C., Martinez-Esparza M., Perez C., Daum N., Solano F., Garcia-Borron J.C. Inhibition of melanogenesis in response to oxidative stress: transient downregulation of melanocyte differentiation markers and possible involvement of microphthalmia transcription factor. J. Cell Sci. 2001; 114(12): 2335–44.Osredkar J., Sustar N. Copper and Zinc, Biological Role and Significance of Copper/Zinc Imbalance. J. Clin. Toxicol. 2011; S3.Osredkar J., Sustar N. Copper and Zinc, Biological Role and Significance of Copper/Zinc Imbalance. J. Clin. Toxicol. 2011; S3.Chai F., Truong-Tran A.Q., Evdokiou A., Young G.P., Zalewski P.D. Intracellular zinc depletion induces caspase activation and p21 Waf1/Cip1 cleavage in human epithelial cell lines. J. Infect. Dis. 2000; 182(1): S85-92.Chai F., Truong-Tran A.Q., Evdokiou A., Young G.P., Zalewski P.D. Intracellular zinc depletion induces caspase activation and p21 Waf1/Cip1 cleavage in human epithelial cell lines. J. Infect. Dis. 2000; 182(1): S85–92.

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