КЛИНИЧЕСКАЯ ЦИТОЛОГИЯ ПЕЧЕНИ: МИТОХОНДРИИ
Ключевые слова:
печень, митохондрии, фиксация, изменения, визуализация
Аннотация
Введение. В последние годы изменились представления о митохондриях (МХ), ранее рассматриваемых только в качестве «энергетических блоков» клетки, вырабатывающих АТФ. МХ являются одной из мишеней атаки HCV при хроническом гепатите С (ХГС). Цель исследования – представить морфологическую характеристику МХ печени при ХГС с применением оригинальной методики фиксации ткани. Материал и методы. Использованы методы световой микроскопии полутонких срезов по усовершенствованной методике фиксации и электронной микроскопии ультратонких срезов. Для получения снимков применены комплекс из цифровой камеры и программа для обработки изображений Olympus Mega View III (Германия). Результаты. В результате исследований выделены следующие варианты изменений МХ в гепатоцитах: набухание, конденсация (уплотнение) матрикса, нарушение структуры крист; нарушение митохондриального слияния; митохондриального деления и фрагментация; разрушение МХ путем образования митофаголизосомы. Приведены иллюстрации макроаутофагии, при которой происходит деградация крупных органелл клетки и крупных белковых агрегатов с образованием аутофагосомы. Изучены инициирующие звенья запуска митохондриального пути апоптоза у ВИЧ-инфицированных пациентов с ХГС, находящихся на антиретровирусной терапии. Заключение. Визуализация МХ в биоптате печени, распределение их в цитоплазме гепатоцита, количество, вариабельность и мониторинг изменений – важные критерии оценки функционального состояния печени в условиях вирусного поражения. Осмиевый фиксатор имеет преимущества перед существующими реагентами.Литература
1. Scheffler IE. Mitochondria. New York: Wiley-Liss; 1999. 480 р.
2. Yoshimitsu K, Hiromi N. Intra-and Intercellular Quality Control Mechanisms of Mitochondria. Cells. 2017;7(1):1. doi: 10.3390/cells7010001.
3. Nelson N, Schatz G. Energy-dependent processing of cytoplasmically made precursors to mitochondrial proteins. Proc. Natl. Asad. Sci. USA. 1979;76(9):4365-4369.
4. Van der Bliek AM, Sedensky MM, Morgan PG. Cell Biology of the Mitochondrion. Genetics. 2017;207(3):843-871. doi: 10.1534/genetics.117.300262.
5. Vakifahmetoglu-Norberg H, Ouchida AT, Norberg E. The role of mitochondria in metabolism and cell death. Biochem. Biophys. Res. Commun. 2017;482:426-431. doi: 10.1016/j.bbrc.2016.11.088.
6. Kiriyma Y, Nochi H. Intra- and Intercellular Quality Control Mechanisms of Mitochondria. Cells. 2018;7(1):1. 7. Lyu BN, Lyu MB, Ismailov BI. Rol mitohondrij v razvitii i regulyacii urovnya okislitelnogo stressa v norme, pri kletochnyh patologiyah i reversii opuholevyh kletok. Advances in Modern Biology. 2006;126(4):388-398. (Russian).
8. Yoshimitsu K, Hiromi N. The Function of Autophagy in Neurodegenerative Diseases. Int. J. Mol. Sci. 2015;16(11):26797-26812.
9. Pan D, Lindau C, Lagies S, Wiedemann N, Kammerer B. Metabolic profiling of isolated mitochondria and cytoplasm reveals compartment-specific metabolic responses. Metabolomics. 2018;14(5):59. doi: 10.1007/s11306-018-1352-x.
10. Kim SJ, Syed GH, Khan M, Chiu WW, Sohail MA, Gish RG, Siddigui A. Hepatitis C virus triggers mitochondrial fission and attenuates apoptosis to promote viral persistence. Proc. Natl. Acad. Sci. USA. 2014;111(17):6413-6418. doi: 10.1073/pnas.1321114111.
11. Chan DC. Mitochondria: Dynamic organelles in disease, aging, and development. Cell. 006;125:1241-1252. doi: 10.1016/j.cell.2006.06.010.
12. Kurbat MN, Tsyrkunov VM, Kondratovich IA. Gepatotoksichnost startovoj skhemy antiretrovirusnoj terapii VICh-infekcii. Medicinskaja panorama [Medical panorama]. 2015;1:3-6. (Russian).
13. Piccoli C, Scrima R, Quarato G, D’Aprile A, Ripoli M, Lecce L, Boffoli D, Moradpour D, Capitanio N. Hepatitis C virus protein expression causes calcium-mediated mitochondrial bioenergetic dysfunction and nitro-oxidative stress. Hepatology. 2007;46:58-65. doi: 10.1002/hep.21679.
14. Quarato G, Scrima R, Agriesti F, Moradpour D, Capitanio N, Piccoli C. Targeting mitochondria in the infection strategy of the hepatitis C virus. Int. J. Biochem. Cell Biol. 2013;45:156-166. doi: 10.1016/j.biocel.2012.06.008.
15. Wang T, Weinman SA. Interactions between Hepatitis C Virus and Mitochondria: Impact on Pathogenesis and Innate Immunity. Curr. Pathobiol. Rep. 2013;1(3):179-187.
16. Sato T, Takagi I. An electron microscopic study of specimen-fixed for longer periods in phosphate buffered formalin. Journal of Electron Microscopy. 1982;31(4):423-428. doi: 10.1093/oxfordjournals.jmicro.a050388.
17. Andreev VP, Matievskaya NV, Tsyrkunov VM, Khombak VV, inventors. Method for fixing liver biopsy specimens. BY рatent 20209. 2016 Ijun 30. (Russian).
18. Glauert RH. Araldite as embedding medium for electron microscopy. Journal of Biophysical and Biochemical Cytology. 1958;4:409-414.
19. Millonig GA. Advantages of a phosphate buffer for osmium tetroxide solutions in fixation. Journal of Applied Physics. 1961;32:1637-1643.
20. Watson ML. Staining of tissue sections for electron microscopy with heavy metals. Journal of Biophysical and Biochemical Cytology. 1958;4:475-478.
21. Glauert AM, editor. Practical Methods in Electron Microscopy. Vol. 3, pt. 1, Glauert AM. Fixation, degydratation and embedding of biological specimens. New York: American Elsevier; 1975. 207 p.
22. Reynolds ES. The use of lead citrate at high pH as an electronopaque stain in electron microscopy. J. Cell Biol. 1963;17(1):208-212.
23. Schaff Z, Lapis K, Andre J. Study of the tridimensional structure of intramitochondrial crystalline inclusions. J. Microscopie. 1974;20:259-264.
24. Serov VV, Lapish K, Sekamova S, Beketova TP; USSR Academy of Medical Sciences. Morfologicheskaja diagnostika zabolevanij pecheni [Morphological diagnosis of liver diseases]. Moscow: Meditsina; 1989. 336 p. (Russian).
25. Riede U, Sandritter W, Mittermayer С. Circulatory shock: a review. Pathology. 1981;13(2):299-311.
26. Ripoli M, D’Aprile A, Quarato G, Sarasin-Filipowicz M,Gouttenoire J, Scrima R, Cela O, Boffoli D, Heim MH, Moradpour D, Capitanio N, Piccoli C. Hepatitis C Viruslinked mitochondrial dysfunction promotes hypoxia-inducible factor 1 alpha-mediated glycolytic adaptation. J. Virol. 2010;84(1):647-660. doi: 10.1128/JVI.00769-09.
27. Siavoshian S, Abraham JD, Thumann C, Kieny MP, Schuster C. Hepatitis C Virus Core, NS3, NS5A, NS5B Proteins Induce Apoptosis in Mature Dendritic Cells. J. Med. Virol. 2005;75(3):402-411. doi: 10.1002/jmv.20283.
28. Brault C, Levy PL, Bartosch B. Hepatitis C virus-induced mitochondrial dysfunctions. Viruses. 2013;5(3):954-980. doi: 10.3390/v5030954.
29. Ding WX, Li M, Biazik JM, Morgan DG, Guo F, Ni HM, Goheen M, Eskelinen EL, Yin XM. Electron microscopic analysis of a spherical mitochondrial structure. J. Biol. Chem. 2012;287(50):42373-42378. doi: 10.1074/jbc.M112.413674.
30. Westermann B. Mitochondrial fusion and fission in cell life and death. Nat. Rev. Mol. Cell Biol. 2010;11(12):872-884. doi: 10.1038/nrm3013.
31. Chan DC. Mitochondria: dynamic organelles in disease, aging, and development. Cell. 2006;125(7):1241-1252. doi: 10.1016/j.cell.2006.06.010.
32. Fujioka H, Tandler B, Hoppel CL. Mitochondrial division in rat cardiomyocytes: an electron microscope study. Anat. Rec. (Hoboken). 2012;295(9):1455-1461. doi: 10.1002/ar.22523.
33. Fujioka H, Tandler B, Consolo MC, Karnik P. Division of Mitochondria in Cultured Human Fibroblasts. Microsc. Res. Tech. 2013;76(12):1213-1216. doi: 10.1002/jemt.22287.
34. Ding WX, Yin XM. Mitophagy: mechanisms, pathophysiological roles, and analysis. Biol Chem. 2012;393(7):547-564. doi: 10.1515/hsz-2012-0119.
35. Rubinsztein DC, Mariño G, Kroemer G. Autophagy and aging. Cell. 2011;146(5):682-695. doi.org/10.1016/j.cell.2011.07.030.
36. Polla BS, Banzet N, Dall AJ, Patrick AA. Vignola M. Les mitochondries, Carrefour entre vie et mort cellulaire : rôles des protéines de stress et consequences sur l’inflammation. Med. Sci.1998;14(1):18-25. (French).
37. Benali-Furet NL, Chami M, Houel L, De Giorgi F, Vernejoul F, Lagorce D, Buscail L, Bartenschlager R, Ichas F, Rizzuto R, Paterlini-Bréchot P. Hepatitis C virus core triggers apoptosis in liver cells by inducing ER stress and ER calcium depletion. Oncogene. 2005;24(31):4921-4933. doi: 10.1038/sj.onc.1208673.
38. Giacomello M, Pellegrini L. The coming of age of the mitochondria–ER contact: a matter of thickness. Cell Death Differ. 2016;23(9):1417-1427. doi: 10.1038/cdd.2016.52.
39. Pupyshev AB. Reparativnaja autofagija i autofagovaja gibel kletki. Funkcionalnye i reguljatornye aspekty [Reparative autophagy and autophagy death of cells. Functional and regulatory aspects]. Tsitologiya [Cytology]. 2014;56(3):179-189. (Russian).
40. Kurbat MN, Tsyrkunov VM. Vzaimosvjaz mezhdu proapoptoticheskimi faktorami mitohondrialnogo zvena apoptoza pri lekarstvennom gepatite. In: Zharko VI, editor. Dostizhenija medicinskoj nauki Belarusi: recenziruemyj nauchno-prakticheskij ezhegodnik [Accomplishments of Medical Science in Belarus]. Minsk: GU RNMB; 2016. [Internet]. Available from: http://med.by/dmn/book. php?book=16-5_4. (Russian).
41. Ivashkin VT. Mekhanizmy immunnoj tolerantnosti i patologii pecheni. Rossijskij Zhurnal Gastrojenterologii, Gepatologii, Koloproktologii [The Russian Journal of Gastroenterology, Hepatology, Coloproctology]. 2009;19(2):2-13. (Russian).
42. Crow, MT. Hypoxia, BNip3 proteins, and the mitochondrial death pathway in cardiomyocytes. Circ. Res. 2002;91(3):183-185.
43. Moyle G. Mitochondrial toxicity: myths and facts. J. HIV Ther. 2004;9(2):45-47.
2. Yoshimitsu K, Hiromi N. Intra-and Intercellular Quality Control Mechanisms of Mitochondria. Cells. 2017;7(1):1. doi: 10.3390/cells7010001.
3. Nelson N, Schatz G. Energy-dependent processing of cytoplasmically made precursors to mitochondrial proteins. Proc. Natl. Asad. Sci. USA. 1979;76(9):4365-4369.
4. Van der Bliek AM, Sedensky MM, Morgan PG. Cell Biology of the Mitochondrion. Genetics. 2017;207(3):843-871. doi: 10.1534/genetics.117.300262.
5. Vakifahmetoglu-Norberg H, Ouchida AT, Norberg E. The role of mitochondria in metabolism and cell death. Biochem. Biophys. Res. Commun. 2017;482:426-431. doi: 10.1016/j.bbrc.2016.11.088.
6. Kiriyma Y, Nochi H. Intra- and Intercellular Quality Control Mechanisms of Mitochondria. Cells. 2018;7(1):1. 7. Lyu BN, Lyu MB, Ismailov BI. Rol mitohondrij v razvitii i regulyacii urovnya okislitelnogo stressa v norme, pri kletochnyh patologiyah i reversii opuholevyh kletok. Advances in Modern Biology. 2006;126(4):388-398. (Russian).
8. Yoshimitsu K, Hiromi N. The Function of Autophagy in Neurodegenerative Diseases. Int. J. Mol. Sci. 2015;16(11):26797-26812.
9. Pan D, Lindau C, Lagies S, Wiedemann N, Kammerer B. Metabolic profiling of isolated mitochondria and cytoplasm reveals compartment-specific metabolic responses. Metabolomics. 2018;14(5):59. doi: 10.1007/s11306-018-1352-x.
10. Kim SJ, Syed GH, Khan M, Chiu WW, Sohail MA, Gish RG, Siddigui A. Hepatitis C virus triggers mitochondrial fission and attenuates apoptosis to promote viral persistence. Proc. Natl. Acad. Sci. USA. 2014;111(17):6413-6418. doi: 10.1073/pnas.1321114111.
11. Chan DC. Mitochondria: Dynamic organelles in disease, aging, and development. Cell. 006;125:1241-1252. doi: 10.1016/j.cell.2006.06.010.
12. Kurbat MN, Tsyrkunov VM, Kondratovich IA. Gepatotoksichnost startovoj skhemy antiretrovirusnoj terapii VICh-infekcii. Medicinskaja panorama [Medical panorama]. 2015;1:3-6. (Russian).
13. Piccoli C, Scrima R, Quarato G, D’Aprile A, Ripoli M, Lecce L, Boffoli D, Moradpour D, Capitanio N. Hepatitis C virus protein expression causes calcium-mediated mitochondrial bioenergetic dysfunction and nitro-oxidative stress. Hepatology. 2007;46:58-65. doi: 10.1002/hep.21679.
14. Quarato G, Scrima R, Agriesti F, Moradpour D, Capitanio N, Piccoli C. Targeting mitochondria in the infection strategy of the hepatitis C virus. Int. J. Biochem. Cell Biol. 2013;45:156-166. doi: 10.1016/j.biocel.2012.06.008.
15. Wang T, Weinman SA. Interactions between Hepatitis C Virus and Mitochondria: Impact on Pathogenesis and Innate Immunity. Curr. Pathobiol. Rep. 2013;1(3):179-187.
16. Sato T, Takagi I. An electron microscopic study of specimen-fixed for longer periods in phosphate buffered formalin. Journal of Electron Microscopy. 1982;31(4):423-428. doi: 10.1093/oxfordjournals.jmicro.a050388.
17. Andreev VP, Matievskaya NV, Tsyrkunov VM, Khombak VV, inventors. Method for fixing liver biopsy specimens. BY рatent 20209. 2016 Ijun 30. (Russian).
18. Glauert RH. Araldite as embedding medium for electron microscopy. Journal of Biophysical and Biochemical Cytology. 1958;4:409-414.
19. Millonig GA. Advantages of a phosphate buffer for osmium tetroxide solutions in fixation. Journal of Applied Physics. 1961;32:1637-1643.
20. Watson ML. Staining of tissue sections for electron microscopy with heavy metals. Journal of Biophysical and Biochemical Cytology. 1958;4:475-478.
21. Glauert AM, editor. Practical Methods in Electron Microscopy. Vol. 3, pt. 1, Glauert AM. Fixation, degydratation and embedding of biological specimens. New York: American Elsevier; 1975. 207 p.
22. Reynolds ES. The use of lead citrate at high pH as an electronopaque stain in electron microscopy. J. Cell Biol. 1963;17(1):208-212.
23. Schaff Z, Lapis K, Andre J. Study of the tridimensional structure of intramitochondrial crystalline inclusions. J. Microscopie. 1974;20:259-264.
24. Serov VV, Lapish K, Sekamova S, Beketova TP; USSR Academy of Medical Sciences. Morfologicheskaja diagnostika zabolevanij pecheni [Morphological diagnosis of liver diseases]. Moscow: Meditsina; 1989. 336 p. (Russian).
25. Riede U, Sandritter W, Mittermayer С. Circulatory shock: a review. Pathology. 1981;13(2):299-311.
26. Ripoli M, D’Aprile A, Quarato G, Sarasin-Filipowicz M,Gouttenoire J, Scrima R, Cela O, Boffoli D, Heim MH, Moradpour D, Capitanio N, Piccoli C. Hepatitis C Viruslinked mitochondrial dysfunction promotes hypoxia-inducible factor 1 alpha-mediated glycolytic adaptation. J. Virol. 2010;84(1):647-660. doi: 10.1128/JVI.00769-09.
27. Siavoshian S, Abraham JD, Thumann C, Kieny MP, Schuster C. Hepatitis C Virus Core, NS3, NS5A, NS5B Proteins Induce Apoptosis in Mature Dendritic Cells. J. Med. Virol. 2005;75(3):402-411. doi: 10.1002/jmv.20283.
28. Brault C, Levy PL, Bartosch B. Hepatitis C virus-induced mitochondrial dysfunctions. Viruses. 2013;5(3):954-980. doi: 10.3390/v5030954.
29. Ding WX, Li M, Biazik JM, Morgan DG, Guo F, Ni HM, Goheen M, Eskelinen EL, Yin XM. Electron microscopic analysis of a spherical mitochondrial structure. J. Biol. Chem. 2012;287(50):42373-42378. doi: 10.1074/jbc.M112.413674.
30. Westermann B. Mitochondrial fusion and fission in cell life and death. Nat. Rev. Mol. Cell Biol. 2010;11(12):872-884. doi: 10.1038/nrm3013.
31. Chan DC. Mitochondria: dynamic organelles in disease, aging, and development. Cell. 2006;125(7):1241-1252. doi: 10.1016/j.cell.2006.06.010.
32. Fujioka H, Tandler B, Hoppel CL. Mitochondrial division in rat cardiomyocytes: an electron microscope study. Anat. Rec. (Hoboken). 2012;295(9):1455-1461. doi: 10.1002/ar.22523.
33. Fujioka H, Tandler B, Consolo MC, Karnik P. Division of Mitochondria in Cultured Human Fibroblasts. Microsc. Res. Tech. 2013;76(12):1213-1216. doi: 10.1002/jemt.22287.
34. Ding WX, Yin XM. Mitophagy: mechanisms, pathophysiological roles, and analysis. Biol Chem. 2012;393(7):547-564. doi: 10.1515/hsz-2012-0119.
35. Rubinsztein DC, Mariño G, Kroemer G. Autophagy and aging. Cell. 2011;146(5):682-695. doi.org/10.1016/j.cell.2011.07.030.
36. Polla BS, Banzet N, Dall AJ, Patrick AA. Vignola M. Les mitochondries, Carrefour entre vie et mort cellulaire : rôles des protéines de stress et consequences sur l’inflammation. Med. Sci.1998;14(1):18-25. (French).
37. Benali-Furet NL, Chami M, Houel L, De Giorgi F, Vernejoul F, Lagorce D, Buscail L, Bartenschlager R, Ichas F, Rizzuto R, Paterlini-Bréchot P. Hepatitis C virus core triggers apoptosis in liver cells by inducing ER stress and ER calcium depletion. Oncogene. 2005;24(31):4921-4933. doi: 10.1038/sj.onc.1208673.
38. Giacomello M, Pellegrini L. The coming of age of the mitochondria–ER contact: a matter of thickness. Cell Death Differ. 2016;23(9):1417-1427. doi: 10.1038/cdd.2016.52.
39. Pupyshev AB. Reparativnaja autofagija i autofagovaja gibel kletki. Funkcionalnye i reguljatornye aspekty [Reparative autophagy and autophagy death of cells. Functional and regulatory aspects]. Tsitologiya [Cytology]. 2014;56(3):179-189. (Russian).
40. Kurbat MN, Tsyrkunov VM. Vzaimosvjaz mezhdu proapoptoticheskimi faktorami mitohondrialnogo zvena apoptoza pri lekarstvennom gepatite. In: Zharko VI, editor. Dostizhenija medicinskoj nauki Belarusi: recenziruemyj nauchno-prakticheskij ezhegodnik [Accomplishments of Medical Science in Belarus]. Minsk: GU RNMB; 2016. [Internet]. Available from: http://med.by/dmn/book. php?book=16-5_4. (Russian).
41. Ivashkin VT. Mekhanizmy immunnoj tolerantnosti i patologii pecheni. Rossijskij Zhurnal Gastrojenterologii, Gepatologii, Koloproktologii [The Russian Journal of Gastroenterology, Hepatology, Coloproctology]. 2009;19(2):2-13. (Russian).
42. Crow, MT. Hypoxia, BNip3 proteins, and the mitochondrial death pathway in cardiomyocytes. Circ. Res. 2002;91(3):183-185.
43. Moyle G. Mitochondrial toxicity: myths and facts. J. HIV Ther. 2004;9(2):45-47.
Опубликован
2019-02-19
Как цитировать
1.
Андреев ВП, Цыркунов ВМ, Кравчук РИ, Курбат МН. КЛИНИЧЕСКАЯ ЦИТОЛОГИЯ ПЕЧЕНИ: МИТОХОНДРИИ. journalHandG [Интернет]. 19 февраль 2019 г. [цитируется по 21 ноябрь 2024 г.];2(2):143-5. доступно на: http://hepatogastro.grsmu.by/index.php/journalHandG/article/view/78
Раздел
Оригинальные исследования
2.png)
