morphoМорфологические ведомостиMorphological newsletter1812-31712686-8741Private Medical University REAVIZ10.20340/mv-mn.2023.31(1).724morpho-724Research ArticleОРИГИНАЛЬНЫЕ ИССЛЕДОВАНИЯRESEARCH ARTICLESИММУНОГИСТОХИМИЧЕСКАЯ ОЦЕНКА ШАПЕРОН-ИНДУЦИРУЕМОЙ АУТОФАГИИ В РАЗЛИЧНЫХ ОТДЕЛАХ ГОЛОВНОГО МОЗГА ЧЕЛОВЕКА ПРИ СТАРЕНИИIMMUNOHISTOCHEMICAL EVALUATION OF CHAPERONE-INDUCED AUTOPHAGY IN VARIOUS PARTS OF THE HUMAN BRAIN DURING AGINGhttps://orcid.org/0000-0001-5223-9767МагнаеваАлина СтаниславовнаMagnaevaAlina S.

аспирантка лаборатории нейроморфологии

Aspirantin of the Laboratory of Neuromorphology

alinamagnaeva03@gmail.comhttps://orcid.org/0000-0002-8999-9986БараничТатьяна ИвановнаBaranichTat'yana I.

кандидат медицинских наук, научный сотрудник лаборатории нейроморфологии

Candidate of Medical Sciences, Researcher of the Laboratory of Neuromorphology

baranich_tatyana@mail.ruhttps://orcid.org/0000-0001-5222-5322ВоронковДмитрий НиколаевичVoronkovDmitry N.

кандидат медицинских наук, старший научный сотрудник лаборатории нейроморфологии

Candidate of Medical Sciences, Senior Researcher of the Laboratory of Neuromorphology

voronkovdm@gmail.comhttps://orcid.org/0000-0002-8635-0786ГофманАнна АндреевнаGofmanAnna A.

студентка

Studentin

makabi2806@gmail.comhttps://orcid.org/0000-0003-1787-775XГулевскаяТатьяна СергеевнаGulevskayaTat'yana S.

доктор медицинских наук, профессор, главный научный сотрудник лаборатории нейроморфологии

Doctor of Medical Sciences, Professor, Chief Researcher at the Laboratory of Neuromorphology

tsgulevskaya077@gmail.comhttps://orcid.org/0000-0001-8708-6940ГлинкинаВалерия ВладимировнаGlinkinaValeriya V.

доктор медицинских наук, профессор, заведующая кафедрой гистологии, эмбриологии и цитологии лечебного факультета

Doctor of Medical Sciences, Professor, Head of the Department of Histology, Embryology and Cytology

vglinkina@mail.ruhttps://orcid.org/0000-0002-0552-6939СухоруковВладимир СергеевичSukhorukovVladimir S.

доктор медицинских наук, профессор, заведующий лабораторией нейроморфологии

Doctor of Medical Sciences, Professor, Head of the Laboratory of Neuromorphology

sukhorukov@neurology.ruНаучный центр неврологии, МоскваРоссияScientific Center of Neurology, MoscowRussian FederationРоссийский национальный исследовательский медицинский университет имени Н.И. Пирогова, МоскваРоссияPirogov Russian National Research Medical University, MoscowRussian Federation2023300120233112733Copyright © Магнаева А.С., Баранич Т.И., Воронков Д.Н., Гофман А.А., Гулевская Т.С., Глинкина В.В., Сухоруков В.С., 20222022Магнаева А.С., Баранич Т.И., Воронков Д.Н., Гофман А.А., Гулевская Т.С., Глинкина В.В., Сухоруков В.С.Magnaeva A.S., Baranich T.I., Voronkov D.N., Gofman A.A., Gulevskaya T.S., Glinkina V.V., Sukhorukov V.S.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.morpholetter.com/jour/article/view/724

Поддержание функционального пула нейронов при возрастной инволюции головного мозга тесно связано с шаперон-опосредованной аутофагией, основной функцией которой является обеспечение клеточного протеостаза и утилизация нейротоксичных белков. Нарушение этого типа аутофагии лежит в основе патогенеза многих возраст-ассоциированных нейродегенеративных заболеваний. В этом аспекте актуальным представляется изучение взаимосвязи между ключевыми маркерами шаперон-опосредованной аутофагии и их относительным вкладом в старение и нейродегенерацию для определения фармацевтических мишеней. Целью исследования явилось определение уровней экспрессии маркеров шаперон-опосредованной аутофагии - белка теплового шока 70 (HSP70) и лизосом-ассоциированного мембранного белка типа 2А (LAMP2) в нейронах различных зон головного мозга человека при старении. Работа была выполнена на аутопсийном материале пациентов, причины смерти которых не были связаны с неврологическими заболеваниями. Исследовали препараты коры прецентральной извилины, стриатума и гиппокампа в группе случаев молодого возраста (35-45 лет, n=5) и старческого возраста (>75 лет, n=10). Иммуногистохимическое окрашивание выполнялось с использованием первичных антител к HSP70 и LAMP2. Оптическая плотность оценивалась в перикарионе 150 крупных нейронов V слоя коры прецентральной извилины, стриатума, пирамидного слоя гиппокампа. Результаты проведенного исследования продемонстрировали увеличение экспрессии как HSP70, так и LAMP2A в препаратах головного мозга случаев старческого возраста. Выявленные более высокие показатели оптической плотности HSP70 по сравнению с LAMP2A в препаратах головного мозга случаев старческого возраста, вероятно, обусловлены диссоциацией между стадией связывания субстрата и стадией его транслокации. Можно предположить, что связывание нефункционирующих белков с HSP70 и кошаперонами происходит быстрее, чем их перемещение в просвет лизосом посредством LAMP2A. Полученные результаты дают основание предполагать, что повышенные уровни маркеров шаперон-опосредованной аутофагии могут носить компенсаторно-приспособительный характер и обеспечивать выживаемость нейронов в условиях хронического стресса при физиологическом старении.

Supporting of the functional pool of neurons during age-related brain involution is closely related to chaperone-mediated autophagy, the main function of which is to provide cellular proteostasis and utilize neurotoxic proteins. Impairment of this type of autophagy underlies the pathogenesis of many age-associated neurodegenerative diseases. In this aspect, it seems relevant to study the relationship between key markers of chaperone-mediated autophagy and their relative contribution to aging and neurodegeneration in order to determine pharmaceutical targets. The aim of the study was to determine the levels of expression of markers of chaperone-mediated autophagy - heat shock protein 70 (HSP70) and lysosome-associated membrane protein type 2A (LAMP2) in neurons of various areas of the human brain during aging. The study was performed on autopsy material of patients whose causes of death were not associated with neurological diseases. Preparations of the cortex of the precentral gyrus, striatum and hippocampus were studied in the group of cases of young age (35-45 years, n=5) and senile age (>75 years, n=10). Immunohistochemical staining was performed using primary antibodies to HSP70 and LAMP2. Optical density was assessed in the perikaryon of 150 large neurons in layer V of the cortex of the precentral gyrus, striatum, and pyramidal layer of the hippocampus. The results of the study demonstrated an increase in the expression of both HSP70 and LAMP2A in brain preparations from cases of senile age. The higher values of the optical density of HSP70 compared to LAMP2A in brain preparations from senile cases were probably due to dissociation between the stage of substrate binding and the stage of its translocation. It can be assumed that the binding of nonfunctioning proteins to HSP70 and cochaperones occurs faster than their movement into the lumen of lysosomes via LAMP2A. The results obtained suggest that the increasing of levels of markers of chaperone-mediated autophagy may be of a compensatory-adaptive nature and ensure the survival of neurons under conditions of chronic stress during physiological aging.

аутофагияголовной мозгстарениеиммуногистохимияшапероныautophagybrainagingimmunohistochemistrychaperonesReferencesLópez-Otín C, Blasco MA, Partridge L et al. The hallmarks of aging. Cell. 2013;153(6):1194-1217. https://doi.org/10.1016/j.cell.2013.05.039López-Otín C, Blasco MA, Partridge L et al. The hallmarks of aging. Cell. 2013;153(6):1194-1217. https://doi.org/10.1016/j.cell.2013.05.039Farr JN, Almeida M. The Spectrum of Fundamental Basic Science Discoveries Contributing to Organismal Aging. J Bone Miner Res. 2018;33(9):1568-1584. https://doi.org/10.1002/jbmr.3564Farr JN, Almeida M. The Spectrum of Fundamental Basic Science Discoveries Contributing to Organismal Aging. J Bone Miner Res. 2018;33(9):1568-1584. https://doi.org/10.1002/jbmr.3564Zhe Li, Zhenkun Zhang, Yikun Ren et al. Aging and age‐related diseases: from mechanisms to therapeutic strategies. Biogerontology. 2021;22(2):165-187. https://doi.org/ 10.1007/s10522-021-09910-5Zhe Li, Zhenkun Zhang, Yikun Ren et al. Aging and age‐related diseases: from mechanisms to therapeutic strategies. Biogerontology. 2021;22(2):165-187. https://doi.org/ 10.1007/s10522-021-09910-5Loeffler DA. Influence of Normal Aging on Brain Autophagy: A Complex Scenario. Front Aging Neurosci. 2019;11:49. https://doi.org/ 10.3389/fnagi.2019.00049Loeffler DA. Influence of Normal Aging on Brain Autophagy: A Complex Scenario. Front Aging Neurosci. 2019;11:49. https://doi.org/ 10.3389/fnagi.2019.00049Loos B, Klionsky DJ, Wong E. Augmenting brain metabolism to increase macro- and chaperone-mediated autophagy for decreasing neuronal proteotoxicity and aging. Prog Neurobiol. 2017;156:90-106. https://doi.org/ 10.1016/j.pneurobio.2017.05.001Loos B, Klionsky DJ, Wong E. Augmenting brain metabolism to increase macro- and chaperone-mediated autophagy for decreasing neuronal proteotoxicity and aging. Prog Neurobiol. 2017;156:90-106. https://doi.org/ 10.1016/j.pneurobio.2017.05.001Kallergi E, Nikoletopoulou V. Macroautophagy and normal aging of the nervous system: Lessons from animal models. Cell Stress. 2021;5(10):146-166. https://doi.org/10.15698/cst2021.10.257Kallergi E, Nikoletopoulou V. Macroautophagy and normal aging of the nervous system: Lessons from animal models. Cell Stress. 2021;5(10):146-166. https://doi.org/10.15698/cst2021.10.257Auzmendi-Iriarte J, Matheu A. Impact of Chaperone-Mediated Autophagy in Brain Aging: Neurodegenerative Diseases and Glioblastoma. Front Aging Neurosci. 2021;12:630743. https://doi.org/10.3389/fnagi.2020.630743Auzmendi-Iriarte J, Matheu A. Impact of Chaperone-Mediated Autophagy in Brain Aging: Neurodegenerative Diseases and Glioblastoma. Front Aging Neurosci. 2021;12:630743. https://doi.org/10.3389/fnagi.2020.630743Maksimovich NE, Bon' EI. Belki teplovogo shoka. Svoistva. Rol' v adaptatsii. Metodicheskie podkhody k opredeleniyu. Biomeditsina. 2020;16(2):60-67. In Russian. https://doi.org/10.33647/2074-5982- 16-2-60-67Maksimovich NE, Bon' EI. Belki teplovogo shoka. Svoistva. Rol' v adaptatsii. Metodicheskie podkhody k opredeleniyu. Biomeditsina. 2020;16(2):60-67. In Russian. https://doi.org/10.33647/2074-5982- 16-2-60-67Pokrovskiy VM, Patrakhanov EA, Antsiferov OV i dr. Belok teplovogo shoka HSP70 - predposylki ispol'zovaniya v kachestve lekarstvennogo sredstva. Farmatsiya i farmakologiya. In Russian. 2021;9(5):346-355. https://doi.org/10.19163/2307-9266-2021-9-5-346-355Pokrovskiy VM, Patrakhanov EA, Antsiferov OV i dr. Belok teplovogo shoka HSP70 - predposylki ispol'zovaniya v kachestve lekarstvennogo sredstva. Farmatsiya i farmakologiya. In Russian. 2021;9(5):346-355. https://doi.org/10.19163/2307-9266-2021-9-5-346-355Kanno H, Handa K, Murakami T et al. Chaperone-Mediated Autophagy in Neurodegenerative Diseases and Acute Neurological Insults in the Central Nervous System. Cells. 2022;11(7):1205. https://doi.org/10.3390/cells11071205Kanno H, Handa K, Murakami T et al. Chaperone-Mediated Autophagy in Neurodegenerative Diseases and Acute Neurological Insults in the Central Nervous System. Cells. 2022;11(7):1205. https://doi.org/10.3390/cells11071205Sukhorukov V, Voronkov D, Baranich T et al. Impaired Mitophagy in Neurons and Glial Cells during Aging and Age-Related Disorders. Int J Mol Sci. 2021;22(19):10251. https://doi.org/10.3390/ijms221910251Sukhorukov V, Voronkov D, Baranich T et al. Impaired Mitophagy in Neurons and Glial Cells during Aging and Age-Related Disorders. Int J Mol Sci. 2021;22(19):10251. https://doi.org/10.3390/ijms221910251Franklin TB, Krueger-Naug AM, Clarke DB et al. The role of heat shock proteins Hsp70 and Hsp27 in cellular protection of the central nervous system. Int J Hyperthermia. 2005;21(5):379-392. https://doi.org/10.1080/02656730500069955Franklin TB, Krueger-Naug AM, Clarke DB et al. The role of heat shock proteins Hsp70 and Hsp27 in cellular protection of the central nervous system. Int J Hyperthermia. 2005;21(5):379-392. https://doi.org/10.1080/02656730500069955Bourdenx M, Martín-Segura A, Scrivo A et al. Chaperone-mediated autophagy prevents collapse of the neuronal metastable proteome. Cell. 2021;184(10):2696-2714.e25. https://doi.org/10.1016/j.cell.2021.03.048Bourdenx M, Martín-Segura A, Scrivo A et al. Chaperone-mediated autophagy prevents collapse of the neuronal metastable proteome. Cell. 2021;184(10):2696-2714.e25. https://doi.org/10.1016/j.cell.2021.03.048Ye W, Xu K, Huang D et al. Age-related increases of macroautophagy and chaperone-mediated autophagy in rat nucleus pulposus. Connect Tissue Res. 2011;52(6):472-478. https://doi.org/ 10.3109/03008207.2011.564336Ye W, Xu K, Huang D et al. Age-related increases of macroautophagy and chaperone-mediated autophagy in rat nucleus pulposus. Connect Tissue Res. 2011;52(6):472-478. https://doi.org/ 10.3109/03008207.2011.564336Salway KD, Gallagher EJ, Page MM, Stuart JA. Higher levels of heat shock proteins in longer-lived mammals and birds. Mech Ageing Dev. 2011;132(6-7):287-297. https://doi.org/10.1016/j.mad.2011.06.002Salway KD, Gallagher EJ, Page MM, Stuart JA. Higher levels of heat shock proteins in longer-lived mammals and birds. Mech Ageing Dev. 2011;132(6-7):287-297. https://doi.org/10.1016/j.mad.2011.06.002Gleixner AM, Pulugulla SH, Pant DB et al. Impact of aging on heat shock protein expression in the substantia nigra and striatum of the female rat. Cell Tissue Res. 2014;357:43–54. https://doi.org/10.1007/s00441-014-1852-6Gleixner AM, Pulugulla SH, Pant DB et al. Impact of aging on heat shock protein expression in the substantia nigra and striatum of the female rat. Cell Tissue Res. 2014;357:43–54. https://doi.org/10.1007/s00441-014-1852-6Joseph AM, Adhihetty PJ, Buford TW et al. The impact of aging on mitochondrial function and biogenesis pathways in skeletal muscle of sedentary high and low-functioning elderly individuals. Aging Cell. 2012;11(5):801–809. https://doi.org/10.1111/j.1474-9726.2012.00844.xJoseph AM, Adhihetty PJ, Buford TW et al. The impact of aging on mitochondrial function and biogenesis pathways in skeletal muscle of sedentary high and low-functioning elderly individuals. Aging Cell. 2012;11(5):801–809. https://doi.org/10.1111/j.1474-9726.2012.00844.xShilova V, Zatsepina O, Zakluta A et al. Age-dependent expression profiles of two adaptogenic systems and thermotolerance in Drosophila melanogaster. Cell Stress Chaperones. 2020;25(2):305-315. https://doi.org/10.1007/s12192-020-01074-4Shilova V, Zatsepina O, Zakluta A et al. Age-dependent expression profiles of two adaptogenic systems and thermotolerance in Drosophila melanogaster. Cell Stress Chaperones. 2020;25(2):305-315. https://doi.org/10.1007/s12192-020-01074-4Cumming KT, Kvamme NH, Schaad L et al. Muscular HSP70 content is higher in elderly compared to young, but is normalized after 12 weeks of strength training. Eur J Appl Physiol. 2021;121(6):1689-1699. https://doi.org/10.1007/s00421-021-04633-4Cumming KT, Kvamme NH, Schaad L et al. Muscular HSP70 content is higher in elderly compared to young, but is normalized after 12 weeks of strength training. Eur J Appl Physiol. 2021;121(6):1689-1699. https://doi.org/10.1007/s00421-021-04633-4Calabrese V, Scapagnini G, Ravagna A, Colombrita C, Spadaro F, Butterfield DA, Giuffrida Stella AM. Increased expression of heat shock proteins in rat brain during aging: relationship with mitochondrial function and glutathione redox state. Mech Ageing Dev. 2004;125(4):325-335. https://doi.org/10.1016/j.mad.2004.01.003Calabrese V, Scapagnini G, Ravagna A, Colombrita C, Spadaro F, Butterfield DA, Giuffrida Stella AM. Increased expression of heat shock proteins in rat brain during aging: relationship with mitochondrial function and glutathione redox state. Mech Ageing Dev. 2004;125(4):325-335. https://doi.org/10.1016/j.mad.2004.01.003Tower J. Hsps and aging. Trends in Endocrinology&Metabolism. 2009;20(5):216-222. https://doi.org/10.1016/j.tem.2008.12.005Tower J. Hsps and aging. Trends in Endocrinology&Metabolism. 2009;20(5):216-222. https://doi.org/10.1016/j.tem.2008.12.005Kikis EA, Gidalevitz T, Morimoto RI. Protein homeostasis in models of aging and age-related conformational disease. Adv Exp Med Biol. 2010;694:138-159. https://doi.org/10.1007/978-1-4419-7002-2_11.Kikis EA, Gidalevitz T, Morimoto RI. Protein homeostasis in models of aging and age-related conformational disease. Adv Exp Med Biol. 2010;694:138-159. https://doi.org/10.1007/978-1-4419-7002-2_11.Njemini R, Lambert M, Demanet C et al. Basal and infection-induced levels of heat shock proteins in human aging. Biogerontology. 2007;8(3):353-364. https://doi.org/ 10.1007/s10522-006-9078-yNjemini R, Lambert M, Demanet C et al. Basal and infection-induced levels of heat shock proteins in human aging. Biogerontology. 2007;8(3):353-364. https://doi.org/ 10.1007/s10522-006-9078-y

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