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Результаты пилотного исследования микроРНК у больных с ишемическим инсультом
Результаты пилотного исследования микроРНК у больных с ишемическим инсультом
Лянг О.В., Кочетов А.Г., Гимадиев Р.Р. и др. Результаты пилотного исследования микроРНК у больных с ишемическим инсультом. Consilium Medicum. 2018; 20 (9): 8–11. DOI: 10.26442/2075-1753_2018.9.8-11
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Аннотация
Актуальность. Высокая медико-социальная значимость ишемического инсульта (ИИ) обусловливает поиск новых диагностических и прогностических биомаркеров, которыми могут стать микроРНК – некодирующие РНК малой длины, супрессирующие экспрессию белок-кодирующих генов. До настоящего момента исследований уровней и роли микроРНК у больных с ИИ в России не проводилось.
Результаты. Статистически значимая динамика к 4-м суткам наблюдения у больных с ИИ была выявлена по уровням микроРНК-125 в плазме, микроРНК-126 в соскобе и микроРНК-145 в соскобе. Также были выявлены статистически значимые различия по уровню в соскобе и плазме микроРНК-126 на 1 и 4-е сутки, а также микроРНК-125 на 4-е сутки наблюдения. По развитию летального исхода были выявлены статистически значимые различия по уровню микроРНК-125 в буккальном соскобе на 1-е сутки, микроРНК-145 в буккальном соскобе на 1-е сутки и микроРНК-21 в плазме на 1-е сутки наблюдения. Также были выявлены различия по таким осложнениям, как пневмония, тромбоэмболия легочной артерии, пиелонефрит. Статистически значимых различий в уровнях микроРНК по типам ИИ, а также по наличию и типу геморрагической трансформации зафиксировано не было.
The aim was to determine the levels of microRNA-21, 125, 126, 145 in plasma and buccal scraping in patients with ischemic stroke.
Materials and methods. The study included 36 patients with acute ischemic stroke. A biomaterial for the study of microRNA-21, 125, 126, 145 in EDTA-plasma and bukkalno scrapings were taken on the 1st and 4th day from the beginning of the development of the disease. Determination of the level of microRNA included the stages of isolation, reverse transcription and real-time PCR. Statistical processing of the study data was carried out using software SPSS 8.0, Microsoft Excel 2013.
Results. Statistically significant dynamics by 4 days of observation in patients with AI was revealed by levels of microRNA-125 in plasma, microRNA-126 in scraping and microRNA-145 in scraping. There were also statistically significant differences in the level of microRNA-126 in 1 and 4 days, and microRNA-125 in 4 days of observation. The development of the lethal outcome revealed statistically significant differences in the level of miRNA-125 in buccal scraping on 1 day, miRNA-145 in buccal scraping on 1 day and microRNA-21 in plasma on 1 day of observation. Also, the differences in such complications as pneumonia, pulmonary embolism, pyelonephritis. There were no statistically significant differences in the levels of miRNAs by type of AI, as well as by the presence and type of hemorrhagic transformation.
Conclusion. MicroRNA-21, 125, 145 for 1 day of observation from the development of AI may have significance in the prognosis of fatal outcome, microRNA-21, 125 for 1 day in the forecast for the development of pneumonia, microRNA-125 for 1 day – the forecast PE, microRNA-126 on the 4th day – in the prediction of pyelonephritis. The revealed absence of differences in levels of microRNA-21 and 125 in scraping and plasma is a possible basis for the application of a non-invasive method of taking biomaterial for the study of microRNA.
Key words: ischemic stroke, microRNA, biomarkers.
Цель – определение уровней микроРНК-21, 125, 126, 145 в плазме и буккальном соскобе у больных с ИИ.
Материалы и методы. В исследование были включены 36 пациентов в острейшем периоде ИИ. Биоматериал для исследования микроРНК-21, 125, 126, 145 в ЭДТА-плазме и буккальном соскобе забирали на 1 и 4-е сутки от начала развития заболевания. Определение уровня микроРНК включало этапы выделения, обратной транскрипции и полимеразной цепной реакции в режиме реального времени. Статистическая обработка данных исследования проведена с использованием программного обеспечения SPSS 8.0, Microsoft Excel 2013.Результаты. Статистически значимая динамика к 4-м суткам наблюдения у больных с ИИ была выявлена по уровням микроРНК-125 в плазме, микроРНК-126 в соскобе и микроРНК-145 в соскобе. Также были выявлены статистически значимые различия по уровню в соскобе и плазме микроРНК-126 на 1 и 4-е сутки, а также микроРНК-125 на 4-е сутки наблюдения. По развитию летального исхода были выявлены статистически значимые различия по уровню микроРНК-125 в буккальном соскобе на 1-е сутки, микроРНК-145 в буккальном соскобе на 1-е сутки и микроРНК-21 в плазме на 1-е сутки наблюдения. Также были выявлены различия по таким осложнениям, как пневмония, тромбоэмболия легочной артерии, пиелонефрит. Статистически значимых различий в уровнях микроРНК по типам ИИ, а также по наличию и типу геморрагической трансформации зафиксировано не было.
Выводы. МикроРНК-21, 125, 145 на 1-е сутки наблюдения от развития ИИ могут иметь значимость в прогнозе летального исхода, микроРНК-21, 125 на 1-е сутки – в прогнозе развития пневмонии, микроРНК-125 на 1-е сутки – тромбоэмболии легочной артерии, микроРНК-126 на 4-е сутки – пиелонефрита. Выявленное отсутствие различий в уровнях микроРНК-21 и 125 в соскобе и плазме представляет собой возможную основу для применения неинвазивного метода взятия биоматериала с целью исследования микроРНК.
Ключевые слова: ишемический инсульт, микроРНК, биомаркеры.
Ключевые слова: ишемический инсульт, микроРНК, биомаркеры.
________________________________________________
The aim was to determine the levels of microRNA-21, 125, 126, 145 in plasma and buccal scraping in patients with ischemic stroke.
Materials and methods. The study included 36 patients with acute ischemic stroke. A biomaterial for the study of microRNA-21, 125, 126, 145 in EDTA-plasma and bukkalno scrapings were taken on the 1st and 4th day from the beginning of the development of the disease. Determination of the level of microRNA included the stages of isolation, reverse transcription and real-time PCR. Statistical processing of the study data was carried out using software SPSS 8.0, Microsoft Excel 2013.
Results. Statistically significant dynamics by 4 days of observation in patients with AI was revealed by levels of microRNA-125 in plasma, microRNA-126 in scraping and microRNA-145 in scraping. There were also statistically significant differences in the level of microRNA-126 in 1 and 4 days, and microRNA-125 in 4 days of observation. The development of the lethal outcome revealed statistically significant differences in the level of miRNA-125 in buccal scraping on 1 day, miRNA-145 in buccal scraping on 1 day and microRNA-21 in plasma on 1 day of observation. Also, the differences in such complications as pneumonia, pulmonary embolism, pyelonephritis. There were no statistically significant differences in the levels of miRNAs by type of AI, as well as by the presence and type of hemorrhagic transformation.
Conclusion. MicroRNA-21, 125, 145 for 1 day of observation from the development of AI may have significance in the prognosis of fatal outcome, microRNA-21, 125 for 1 day in the forecast for the development of pneumonia, microRNA-125 for 1 day – the forecast PE, microRNA-126 on the 4th day – in the prediction of pyelonephritis. The revealed absence of differences in levels of microRNA-21 and 125 in scraping and plasma is a possible basis for the application of a non-invasive method of taking biomaterial for the study of microRNA.
Key words: ischemic stroke, microRNA, biomarkers.
Полный текст
Список литературы
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20. Schwarzenbach H, da Silva AM, Calin G, Pantel K. Data Normalization Strategies for MicroRNA Quantification. Clin Chem 2015; 61 (11): 1333–42.
21. Schwarzenbach H, da Silva AM, Calin G, Pantel K. Which is the accurate data normalization strategy for microRNA quantification? Clin Chem 2015; 61 (11): 1333–42.
22. Zampetaki A, Willeit P, Drozdov I et al. Profiling of circulating microRNAs: from single biomarkers to re-wired networks. Cardiovasc Res 2012; 93 (4): 555–62.
2. Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 2004; 116 (2): 281–97.
3. Chen X, Ba Y, Ma L et al. Characterization of microRNAs in serum: a novel class of biomarkers for diagnosis of cancer and other diseases. Cell Res 2008; 18: 997–1006.
4. Pritchard CC, Kroh E, Wood B et al. Blood cell origin of circulating microRNAs: a cautionary note for cancer biomarker studies. Cancer Prev Res (Phila) 2012; 5: 492–7.
5. Wang K, Zhang S, Weber J et al. Export of microRNAs and microRNA-protective protein by mammalian cells. Nucleic Acids Res 2010; 38: 7248–59.
6. Rainen L, Oelmueller U, Jurgensen S et al. Stabilization of mRNA expression in whole blood samples. Clin Chem 2002; 48: 1883–90.
7. Ai J, Zhang R, Li Y et al. Circulating microRNA-1 as a potential novel biomarker for acute myocardial infarction. Biochem Biophys Res Commun 2010; 391: 73–7.
8. Thum T, Gross C, Fiedler J et al. MicroRNA-21 contributes to myocardial disease by stimulating MAP kinase signalling in fibroblasts. Nature 2008; 456: 980–4.
9. Dong S, Ma W, Hao B et al. microRNA-21 promotes cardiac fibrosis and development of heart failure with preserved left ventricular ejection fraction by up-regulating Bcl-2. Int J Clin Exp Pathol 2014; 7 (2): 565–74.
10. Sheedy F, Palsson-McDermott E, Hennessy E et al. Negative regulation of TLR4 via targeting of the proinflammatory tumor suppressor PDCD4 by the microRNA miR-21. Nat Immunol 2010; 11: 141–7.
11. Wojtowicz EE, Walasek MA, Broekhuis MJ et al. MicroRNA-125 family members exert a similar role in the regulation of murine hematopoiesis. Exp Hematol 2014; 42 (10): 909–18.
12. Le MT, Shyh-Chang N, Khaw SL et al. Conserved regulation of p53 network dosage by microRNA-125b occurs through evolving miRNA-target gene pairs. PLoS Genet 2011; 7 (9): e1002242.
13. Rink C, Khanna S. MicroRNA in ischemic stroke etiology and pathology. Physiol Genomics 2011; 43 (10): 521–8.
14. Ishizaki T, Tamiya T, Taniguchi K et al. miR126 positively regulates mast cell proliferation and cytokine production through suppressing Spred1. Genes Cells 2011; 16: 803–14.
15. Fichtlscherer S, De Rosa S, Fox H, Schwietz T. Circulating microRNAs in patients with coronary artery disease. Circ Res 2010; 107 (5): 677–84.
16. Jia L, Hao F, Wang W, Qu Y. Circulating miR-145 is associated with plasma high-sensitivity C-reactive protein in acute ischemic stroke patients. Cell Biochem Funct 2015; 33 (5): 314–9.
17. Dong YM, Liu XX, Wei GQ et al. Prediction of long-term outcome after acute myocardial infarction using circulating miR-145. Scand J Clin Lab Invest 2015; 75 (1): 85–91.
18. Shliakhto E.V., Barantsevich E.R., Shcherbak N.S., Galagudza M.M. Molekuliarnye mekhanizmy formirovaniia ishemicheskoi tolerantnosti golovnogo mozga (obzor literatury. Chast' 2). Vestn. RAMN. 2012; 7: 20–9. [in Russian]
19. Novikova L.B., Minibaeva G.M. Rol' mikroRNK v patogeneze ishemicheskogo insul'ta. Zhurn. nevrologii i psikhiatrii im. C.C.Korsakova. 2018; 118 (2–3): 43–7. [in Russian]
20. Schwarzenbach H, da Silva AM, Calin G, Pantel K. Data Normalization Strategies for MicroRNA Quantification. Clin Chem 2015; 61 (11): 1333–42.
21. Schwarzenbach H, da Silva AM, Calin G, Pantel K. Which is the accurate data normalization strategy for microRNA quantification? Clin Chem 2015; 61 (11): 1333–42.
22. Zampetaki A, Willeit P, Drozdov I et al. Profiling of circulating microRNAs: from single biomarkers to re-wired networks. Cardiovasc Res 2012; 93 (4): 555–62.
2. Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 2004; 116 (2): 281–97.
3. Chen X, Ba Y, Ma L et al. Characterization of microRNAs in serum: a novel class of biomarkers for diagnosis of cancer and other diseases. Cell Res 2008; 18: 997–1006.
4. Pritchard CC, Kroh E, Wood B et al. Blood cell origin of circulating microRNAs: a cautionary note for cancer biomarker studies. Cancer Prev Res (Phila) 2012; 5: 492–7.
5. Wang K, Zhang S, Weber J et al. Export of microRNAs and microRNA-protective protein by mammalian cells. Nucleic Acids Res 2010; 38: 7248–59.
6. Rainen L, Oelmueller U, Jurgensen S et al. Stabilization of mRNA expression in whole blood samples. Clin Chem 2002; 48: 1883–90.
7. Ai J, Zhang R, Li Y et al. Circulating microRNA-1 as a potential novel biomarker for acute myocardial infarction. Biochem Biophys Res Commun 2010; 391: 73–7.
8. Thum T, Gross C, Fiedler J et al. MicroRNA-21 contributes to myocardial disease by stimulating MAP kinase signalling in fibroblasts. Nature 2008; 456: 980–4.
9. Dong S, Ma W, Hao B et al. microRNA-21 promotes cardiac fibrosis and development of heart failure with preserved left ventricular ejection fraction by up-regulating Bcl-2. Int J Clin Exp Pathol 2014; 7 (2): 565–74.
10. Sheedy F, Palsson-McDermott E, Hennessy E et al. Negative regulation of TLR4 via targeting of the proinflammatory tumor suppressor PDCD4 by the microRNA miR-21. Nat Immunol 2010; 11: 141–7.
11. Wojtowicz EE, Walasek MA, Broekhuis MJ et al. MicroRNA-125 family members exert a similar role in the regulation of murine hematopoiesis. Exp Hematol 2014; 42 (10): 909–18.
12. Le MT, Shyh-Chang N, Khaw SL et al. Conserved regulation of p53 network dosage by microRNA-125b occurs through evolving miRNA-target gene pairs. PLoS Genet 2011; 7 (9): e1002242.
13. Rink C, Khanna S. MicroRNA in ischemic stroke etiology and pathology. Physiol Genomics 2011; 43 (10): 521–8.
14. Ishizaki T, Tamiya T, Taniguchi K et al. miR126 positively regulates mast cell proliferation and cytokine production through suppressing Spred1. Genes Cells 2011; 16: 803–14.
15. Fichtlscherer S, De Rosa S, Fox H, Schwietz T. Circulating microRNAs in patients with coronary artery disease. Circ Res 2010; 107 (5): 677–84.
16. Jia L, Hao F, Wang W, Qu Y. Circulating miR-145 is associated with plasma high-sensitivity C-reactive protein in acute ischemic stroke patients. Cell Biochem Funct 2015; 33 (5): 314–9.
17. Dong YM, Liu XX, Wei GQ et al. Prediction of long-term outcome after acute myocardial infarction using circulating miR-145. Scand J Clin Lab Invest 2015; 75 (1): 85–91.
18. Шляхто Е.В., Баранцевич Е.Р., Щербак Н.С., Галагудза М.М. Молекулярные механизмы формирования ишемической толерантности головного мозга (обзор литературы. Часть 2). Вестн. РАМН. 2012; 7: 20–9. / Shliakhto E.V., Barantsevich E.R., Shcherbak N.S., Galagudza M.M. Molekuliarnye mekhanizmy formirovaniia ishemicheskoi tolerantnosti golovnogo mozga (obzor literatury. Chast' 2). Vestn. RAMN. 2012; 7: 20–9. [in Russian]
19. Новикова Л.Б., Минибаева Г.М. Роль микроРНК в патогенезе ишемического инсульта. Журн. неврологии и психиатрии им. C.C.Корсакова. 2018; 118 (2–3): 43–7. / Novikova L.B., Minibaeva G.M. Rol' mikroRNK v patogeneze ishemicheskogo insul'ta. Zhurn. nevrologii i psikhiatrii im. C.C.Korsakova. 2018; 118 (2–3): 43–7. [in Russian]
20. Schwarzenbach H, da Silva AM, Calin G, Pantel K. Data Normalization Strategies for MicroRNA Quantification. Clin Chem 2015; 61 (11): 1333–42.
21. Schwarzenbach H, da Silva AM, Calin G, Pantel K. Which is the accurate data normalization strategy for microRNA quantification? Clin Chem 2015; 61 (11): 1333–42.
22. Zampetaki A, Willeit P, Drozdov I et al. Profiling of circulating microRNAs: from single biomarkers to re-wired networks. Cardiovasc Res 2012; 93 (4): 555–62.
________________________________________________
2. Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 2004; 116 (2): 281–97.
3. Chen X, Ba Y, Ma L et al. Characterization of microRNAs in serum: a novel class of biomarkers for diagnosis of cancer and other diseases. Cell Res 2008; 18: 997–1006.
4. Pritchard CC, Kroh E, Wood B et al. Blood cell origin of circulating microRNAs: a cautionary note for cancer biomarker studies. Cancer Prev Res (Phila) 2012; 5: 492–7.
5. Wang K, Zhang S, Weber J et al. Export of microRNAs and microRNA-protective protein by mammalian cells. Nucleic Acids Res 2010; 38: 7248–59.
6. Rainen L, Oelmueller U, Jurgensen S et al. Stabilization of mRNA expression in whole blood samples. Clin Chem 2002; 48: 1883–90.
7. Ai J, Zhang R, Li Y et al. Circulating microRNA-1 as a potential novel biomarker for acute myocardial infarction. Biochem Biophys Res Commun 2010; 391: 73–7.
8. Thum T, Gross C, Fiedler J et al. MicroRNA-21 contributes to myocardial disease by stimulating MAP kinase signalling in fibroblasts. Nature 2008; 456: 980–4.
9. Dong S, Ma W, Hao B et al. microRNA-21 promotes cardiac fibrosis and development of heart failure with preserved left ventricular ejection fraction by up-regulating Bcl-2. Int J Clin Exp Pathol 2014; 7 (2): 565–74.
10. Sheedy F, Palsson-McDermott E, Hennessy E et al. Negative regulation of TLR4 via targeting of the proinflammatory tumor suppressor PDCD4 by the microRNA miR-21. Nat Immunol 2010; 11: 141–7.
11. Wojtowicz EE, Walasek MA, Broekhuis MJ et al. MicroRNA-125 family members exert a similar role in the regulation of murine hematopoiesis. Exp Hematol 2014; 42 (10): 909–18.
12. Le MT, Shyh-Chang N, Khaw SL et al. Conserved regulation of p53 network dosage by microRNA-125b occurs through evolving miRNA-target gene pairs. PLoS Genet 2011; 7 (9): e1002242.
13. Rink C, Khanna S. MicroRNA in ischemic stroke etiology and pathology. Physiol Genomics 2011; 43 (10): 521–8.
14. Ishizaki T, Tamiya T, Taniguchi K et al. miR126 positively regulates mast cell proliferation and cytokine production through suppressing Spred1. Genes Cells 2011; 16: 803–14.
15. Fichtlscherer S, De Rosa S, Fox H, Schwietz T. Circulating microRNAs in patients with coronary artery disease. Circ Res 2010; 107 (5): 677–84.
16. Jia L, Hao F, Wang W, Qu Y. Circulating miR-145 is associated with plasma high-sensitivity C-reactive protein in acute ischemic stroke patients. Cell Biochem Funct 2015; 33 (5): 314–9.
17. Dong YM, Liu XX, Wei GQ et al. Prediction of long-term outcome after acute myocardial infarction using circulating miR-145. Scand J Clin Lab Invest 2015; 75 (1): 85–91.
18. Shliakhto E.V., Barantsevich E.R., Shcherbak N.S., Galagudza M.M. Molekuliarnye mekhanizmy formirovaniia ishemicheskoi tolerantnosti golovnogo mozga (obzor literatury. Chast' 2). Vestn. RAMN. 2012; 7: 20–9. [in Russian]
19. Novikova L.B., Minibaeva G.M. Rol' mikroRNK v patogeneze ishemicheskogo insul'ta. Zhurn. nevrologii i psikhiatrii im. C.C.Korsakova. 2018; 118 (2–3): 43–7. [in Russian]
20. Schwarzenbach H, da Silva AM, Calin G, Pantel K. Data Normalization Strategies for MicroRNA Quantification. Clin Chem 2015; 61 (11): 1333–42.
21. Schwarzenbach H, da Silva AM, Calin G, Pantel K. Which is the accurate data normalization strategy for microRNA quantification? Clin Chem 2015; 61 (11): 1333–42.
22. Zampetaki A, Willeit P, Drozdov I et al. Profiling of circulating microRNAs: from single biomarkers to re-wired networks. Cardiovasc Res 2012; 93 (4): 555–62.
Авторы
О.В.Лянг*1,2, А.Г.Кочетов1–3, Р.Р.Гимадиев4, А.А.Абрамов2, Н.А.Шамалов1, Л.В.Стаховская1
1 ФГБОУ ВО «Российский национальный исследовательский медицинский университет им. Н.И.Пирогова» Минздрава России. 117997, Россия, Москва, ул. Островитянова, д. 1;
2 ФГАОУ ВО «Российский университет дружбы народов». 117198, Россия, Москва, ул. Миклухо-Маклая, д. 6;1 ФГБОУ ВО «Российский национальный исследовательский медицинский университет им. Н.И.Пирогова» Минздрава России. 117997, Россия, Москва, ул. Островитянова, д. 1;
3 ФГБУ «Научный медицинский исследовательский центр кардиологии» Минздрава России. 121552, Россия, Москва, ул. 3-я Черепковская, д. 15А;
4 АНО ДПО «Институт лабораторной медицины». 125315, Россия, Москва, Ленинградский пр-т, д. 80Г, пом. XI
*ov_lyang@dpo-ilm.ru
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1 N.I.Pirogov Russian National Research Medical University of the Ministry of Health of the Russian Federation. 117997, Russian Federation, Moscow, ul. Ostrovitianova, d. 1;
2 People’s Friendship University of Russia. 117198, Russian Federation, Moscow, ul. Miklukho-Maklaya, d. 6;
3 National Medical Research Center for Cardiology of the Ministry of Health of the Russian Federation. 121552, Russian Federation, Moscow, ul. 3-ia Cherepkovskaia, d. 15A;
4 Institute of Laboratory Medicine. 125315, Russian Federation, Moscow, Leningradskiy pr-t, d. 80G, pom. XI
*ov_lyang@dpo-ilm.ru
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