Analysis of annexin V levels and cytokine status in patients with acute myocardial infarction

   Aim. Study of annexin V levels and cytokine status in patients with acute myocardial infarction.
   Materials and methods. We examined 27 patients with Q-forming myocardial infarction who were admitted 12–24 hours after the development of the clinic, and 20 individuals in the control group. The lipid spectrum was determined using an automatic analyzer Humalizer-3000 (Human, Germany). Immunological studies were performed on a flow cytofluorimeter Navios (Beckman Coulter, USA). Lymphocyte subpopulations were evaluated using monoclonal antibodies produced by Immunotech (Coulter Corporation, USA). A set of ANNEXIN V-FITC/7 AAD (Beckman Coulter, USA) was used to isolate cells that had undergone apoptosis. Isolation of peripheral blood mononuclears was performed by centrifugation on a density gradient Ficol-hypac (Pharmacia Fine Chemicals, Sweden), interleukin-1β was determined using the Vector-Best test system (Vector-Best, Russia), and the tumor necrosis factor was determined using the RayBio Human TNF-alpha ELISA Kit (USA). Statistical data processing was performed using the program package Statistica 10.0.
   Results. In patients with acute myocardial infarction on the background of dyslipidemia (total cholesterol and LDL cholesterol increased, cholesterol-HDL reduced) showed a significant decrease in the total number of T-lymphocytes (СD 3+) (p < 0.05), T-suppressor (СD 8+) (p < 0.01), B-cells number (СD 19+) and the ratio of CD 4+/CD 8+ lymphocytes increased (p < 0.05). In patients with myocardial infarction there was a significant increase of apoptotic cells and рro-inflammatory cytokines involved in apoptosis (IL-1β and TNF-α) in the blood compared with the control.
   Conclusion. Increased spontaneous apoptosis of peripheral blood lymphocytes in patients with acute myocardial infarction indicates the systemic nature of apoptosis in acute coronary pathology, due to increased annexin-dependent apoptosis of T-lymphocytes as a result of immune disorders, which leads to activation of the cytokine system with an increased concentration of рro-inflammatory mediators (TNF-α, IL-1β). The identified dependencies require further study with the prospect of developing predictive algorithms for myocardial infarction outcomes based on indicators of annexin V and pro-inflammatory cytokines.

1. Петрищев Н. Н. Содержание растворимых маркеров апоптоза и циркулирующих аннексин V-связанных апоптотических клеток в крови больных острым коронарным синдромом / Н. Н. Петрищев, Л. В. Васина, А. В. Луговая // Вестник Санкт-Петербургского университета. – 2008. – 11 (1): 14–23. – Petrishchev N. N. The content of soluble markers of apoptosis and circulating annexin V-linked apoptotic cells in the blood of patients with acute coronary syndrome. Bulletin of Saint Petersburg University. 2008; 11 (1): 14–23. (In Russ.).

2. Palojoki M., Saraste A., Eriksson A. et al. Cardiomyocyte apoptosis and ventricular remodeling after myocardial infarction in rats. Am. J. Physiol. Heart Circ. Physiol. 2001; 280 (6): 2726–2731. DOI: 10.1152/ajpheart.2001.280.6.H2726.

3. Nakajima H., Yanase N., Oshima K. et al. Enhanced expression of apoptosis inducing ligand TRAIL in mononuclear cells after myocardial infarction. Jpn. Heart J. 2003; 44: 833–844. DOI: 10.1536/jhj.44.833.

4. Шахнович Р. М. Острый коронарный синдром / Р. М. Шахнович // В кн.: Кардиология: национальное руководство. – М.: ГЭОТАР-Медиа, 2007. – 692 с. – Shakhnovich R. M. Acute coronary syndrome. In book: Cardiology: National guidelines. Moscow: GEOTAR-Media; 2007. 692 с.

5. Piro F. R., di Gioia C. R., Gallo P. et al. Is apoptosis a diagnostic marker of acute myocardial infarction? Arch Pathol Lab Med. 2000; 124: 827–831. DOI: 10.5858/2000-124-0827-IAADMO.

6. Залесский В. Н. Апоптоз при ишемии и реперфузии миокарда / В. Н. Залесский, Т. И. Гавриленко, А. А. Фильченков // Врачебное Дело. – 2002. – 1: 8–15. – Zalessky V. N., Gavrilenko T. I., Filchenkov A. A. Apoptosis in myocardial ischemia and reperfusion. Physician’s Business. 2002; 1: 8–15.

7. Москалева Е. Ю. Возможные механизмы адаптации клетки к повреждениям, индуцирующим программированную гибель. Связь с патологией / Е. Ю. Москалева, С. Е. Севрин // Патологическая физиология и экспериментальная терапия. – 2006. – 2: 2–15. – Moskaleva E. Yu., Sevrin S. E. Possible mechanisms of cell adaptation to damage inducing programmed death. Relationship to pathology. Pathological Physiology and Experimental Therapy. 2006; 2: 2–15.

8. Bennet A. M., van Maarle M. C., Hallqvist J. et al. Association of TNF-alpha serum levels and TNFA promoter polymorphisms with risk of myocardial infarction. Atherosclerosis. 2006; 187 (2): 408–414. DOI: 10.1016/j.atherosclerosis.2005.09.022.

9. Ярилин А. А. Система цитокинов и принципы ее функционирования в норме и при патологии / А. А. Ярилин // Иммунология. – 1997. – 5: 7–13. – Yarilin A. A. Cytokine system and principles of its functioning in norm and pathology. Immunology. 1997; 5: 7–13.

10. Ковальчук Л. В. Система цитокинов / Л. В. Ковальчук, Л. В. Ганковская, Э. И. Рубакова. – М.: Янус-К, 2000. – 64 с. – Kovalchuk L. V., Gankovskaya L. V., Rubakova E. I. Cytokine system. Moscow: Janus-K, 2000. 64 с.

11. Петрищев Н. Н. Аннексин А5 и дисфункция эндотелия / Н. Н. Петрищев, Л. В. Васина // Ученые записки Санкт-Петербургского государственного медицинского университета им. академика И. П. Павлова. – 2004. – XI (3): 45–47. – Petrishchev N. N., Vasina L. V. Annexin A5 and endothelial dysfunction. Scientific Notes of St. Petersburg State Medical University named after I. P. Pavlov. 2004; XI (3): 45–47.

12. Fadok V. A., Volker D. R., Campbell P. A. et al. Exposure of phospatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages. J. Immunol. 1992; 148: 22157–22164.

13. Guillermo Mariño & Guido Kroemer. Mechanisms of apoptotic phosphatidylserine exposure. Cell Research. 2013; 23: 1247–1248. Doi: 10.1038/cr.2013.115.

14. Vanags D. M., Coppola S., Burgess D. H. Protease involvement in fodrin cleavage and phosphatidylserine exposure in apoptosis. Biol. Chem. 1996; 271: 31075–31085. DOI: 10.1074/jbc.271.49.31075

15. Reutelingsperger C. P. M., van Heerde W. L. Annexin V, the regulator of phosphatidylserinecatalyzed infl ammation and coagulation during apoptosis. Cell. Mol. Lafe Sci. 1997; 53: 527–532. https://doi.org/10.1007/s000180050067

16. Кудрявцев И. В. Современные методы и подходы к изучению апоптоза в экспериментальной биологии / И. В. Кудрявцев [и др.] // Медицинская иммунология. – 2012. – 14 (6): 461–482. – Kudryavtsev I. V., Golovkin A. S., Zurochka A. V., Haidukov S. V. Modern methods and approaches to the study of apoptosis in experimental biology. Medical Immunology. 2012; 14 (6): 461–482.

17. Williams R. S. Benjamin I. J. Protective responses in the ischemic myocardium. J. Clin. Invest. 2000; 106: 813–818. https://doi.org/10.1172/JCI11205

18. Хусаинова Л. Н. Клеточные маркеры апоптоза при остром коронарном синдроме / Л. Н. Хусаинова [и др.] // Медицинский вестник Башкортостана. – 2013. – 8 (3): 78–81. – Khusainova L. N., Smakaeva E. R., Sadikova R. I., Mingazetdinova L. N. Cell markers of apoptosis in acute coronary syndrome. Medical Bulletin of Bashkortostan. 2013; 8 (3): 78–81.

19. Тепляков А. Т. Ранние маркеры прогрессирования сердечной недостаточности и апоптоза: роль в прогнозировании риска развития неблагоприятных сердечно-сосудистых событий у больных, перенесших инфаркт миокарда / А. Т. Тепляков [и др.] // Бюллетень сибирской медицины. – 2016. – 15 (1): 37–46. DOI: 10.20538/1682–0363–2016–1–37–46. – Teplyakov A. T., Grakova E. V., Berezikova E. N., Shilov S. N., Kopyeva K. V., Kalyuzhin V. V. Early markers of heart failure progression and apoptosis: role in predicting the risk of adverse cardiovascular events in patients with myocardial infarction. Bulletin of Siberian Medicine. 2016; 15 (1): 37–46. DOI: 10.20538/1682–0363–2016–1–37–46.

20. Saraste A., Pulkki K., Kallajoki M. et al. Apoptosis in human acute myocardial infarction. Circulation 1997; 95: 320–323. http://dx.doi.org/10.1161/01.cir.95.2.320

21. Fliss H., Gattinger D. Apoptosis in ischemic and reperfused rat myocardium. Circ Res. 1996; 79: 949–56. DOI: 10.1161/01.res.79.5.949

22. Константинова Е. В. Особенности иммунного ответа и воспалительной реакции при атеротромботическом инсульте и инфаркте миокарда / Е. В. Константинова [и др.] // Журнал неврологии и психиатрии им. С. С. Корсакова. – 2015. – 115 (12): 48–53. DOI: 10.17116/jnevro201511512248–53. – Konstantinova E. V., Kochetov A. G., Shostak N. A. et al. Features of immune response and inflammatory response in atherothrombotic stroke and myocardial infarction. Journal of Neurology and Psychiatry n. a. S. S. Korsakov. 2015; 115 (12): 48–53. DOI: 10.17116/jnevro201511512248–53.

23. Simak J., Holada K., Vostal J. G. Release of annexin A5-binding membrane microparticles from cultured human umbilical vein endothelial cells after treatment with camptothecin. BMC Cell. Biol. 2002; 3: 11. DOI: 10.1186/1471–2121–3–11.

24. Harada-Shiba M., Kinoshita M., Kamido H., Shimokado K. Oxidized low density lipoprotein induces apoptosis in cultured human umbilical vein endothelial cells by common and unique mechanisms. J. Biol. Chem. 1998; 273: 9681–9687. DOI: 10.1074/jbc.273.16.9681.

25. Портянко А. С. Система Fas / Fas L и ее значение для регуляции взаимоотношений опухоли и иммунной системы при папиллярном раке щитовидной железы у детей и подростков / А. С. Портянко, Е. Д. Черствой // Архив патологии. – 2003. – 65 (4): 18–21. – Portianko A. S., Cherstoy E. D. The Fas / Fas L system and its significance for the regulation of tumor-immune system interactions in papillary thyroid cancer in children and adolescents. Archives of Pathology. 2003; 65 (4): 18–21.

26. Константинова Е. В. Взаимосвязь апоптоза и экспрессии белков теплового шока лимфоцитов периферической крови у больных с инфарктом миокарда / Е. В. Константинова [и др.] // Бюллетень экспериментальной биологии и медицины. – 2010. – 12: 622–625. – Konstantinova E. V., Khomyakova N. F., Konstantinova N. A. et al. Relationship between apoptosis and heat shock protein expression of peripheral blood lymphocytes in patients with myocardial infarction. Bulletin of Experimental Biology and Medicine. 2010; 12: 622–625.

27. Ceconi C., Curello S., Bachetti T. et al. Tumor necrosis factor in congestive heart failure: a mechanism of disease for the new millennium. Prog. Cardiovasc. Dis. 1998; 41 (1): 25–30.

28. Guillen I., Blanes M., Gomez-Lechon M. J., Castell J. V. Cytokine signaling during myocardial infarction: sequential appearance of IL-1 beta and IL-6. Am. J. Physiol. 1995; 269: 229–235. DOI: 10.1152/ajpregu.1995.269.2.R229.