Study of the pharmacological activity of novel EPOR/CD131 heteroreceptor agonists in mice with endothelial-specific expression of mutant Polg gene

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The aim of the research was to study antiatherosclerotic and endothelial kinds of a protective activity of peptides mimicking an erythropoietin a-helix B tertiary structure with laboratory codes EP-11-1 (UEHLERALNSS), EP-11-2. (UEQLERALNCS), EP-11-3 (UEQLERALNTS).

Materials and methods. The study was conducted on 96 C57Bl/6J male double transgenic Polgmut/mut/Cdh5-CRE mice. Atherosclerosis was induced by a balloon injury accompanied by Western diet. Then, for 27 days, the drugs under study were administered once per 3 days at the dose of 20 μg/kg. On the 28th day, the animals were euthanized and the area of atherosclerotic plaques was collected for an assessment. The expression of genes associated with the processes of inflammation, apoptosis, and angiogenesis was determined in the tissues of the aorta. In addition, the endothelial protective effect of peptides in isolated segments of the thoracic aorta of wild and transgenic ransgenic Polgmut/mut mice was studied.

Results. The assessment of the plaque size in the animals with the Polgmut/mut/Cdh5-CRE genotype against the background of the peptides under study did not reveal statistically significant differences in comparison to control. However, a quantitative PCR showed a statistically significant decreased expression of pro-apoptotic factors p-53 and Bax, and also increase the expression of anti-apoptotic factor Bcl-2 against the background of the peptides EP-11-1 and EP-11-2 administration. The administration of EP-11-1 and the original peptide pHBSP resulted in a statistically significant decrease in the Bax/Bcl-2 ratio. Compounds EP-11-1, EP-11-2, and EP-11-3 were more effective than the original peptide pHBSP, in reducing the increased expression of genes for inflammatory markers iNos, intercellular adhesion molecules Icam-1, Vcam-1 and E-selectin. The use of EP-11-1 led to a more efficient, in comparison with pHBSP, restoration of endothelial-dependent vasodilation of the aortic segments in mice with endothelial-specific overexpression of the mutant Polg gene.

Conclusion. The study carried out on a murine model of the endothelial-specific expression of mutant gamma polymerase has shown that derivatives of the pHBSP peptide with laboratory codes EP-11-1, EP-11-2, EP-11-3, obtained by BLAST-searching for groups of pHBSP related peptides, have atheroprotective and endothelial protective kinds of a protective activity, which is more pronounced in comparison with the original peptide pHBSP.

作者简介

Mikhail Korokin

Belgorod State National Research University

编辑信件的主要联系方式.
Email: mkorokin@mail.ru
ORCID iD: 0000-0001-5402-0697

Doctor of Sciences (Medicine), Associate Professor, Head of the Laboratory of the Pharmacology of Living Systems Research Institute

俄罗斯联邦, 85, Pobedа Str., Belgorod, Russia, 308015

Marina Kubekina

Institute of Gene Biology, Russian Academy of Sciences

Email: kubekina@genebiology.ru
ORCID iD: 0000-0002-8834-1111

Postgraduate Student, Junior Researcher

俄罗斯联邦, Bldg. 5, 34, Vavilov Str., Moscow, Russia, 119334

Alexey Deykin

Belgorod State National Research University; Institute of Gene Biology, Russian Academy of Sciences

Email: alexei@deikin.ru
ORCID iD: 0000-0001-9960-0863

Candidate of Sciences (Biology)

俄罗斯联邦, 85, Pobedа Str., Belgorod, Russia, 308015; Bldg. 5, 34, Vavilov Str., Moscow, Russia, 119334

Oleg Antsiferov

Belgorod State National Research University

Email: antsiferov@bsu.edu.ru
ORCID iD: 0000-0001-6439-2419

Senior Lecturer of the Departament of Faculty Therapy

俄罗斯联邦, 85, Pobedа Str., Belgorod, Russia, 308015

Vladimir Pokrovskii

Belgorod State National Research University

Email: vmpokrovsky@yandex.ru
ORCID iD: 0000-0003-3138-2075

6th year student

俄罗斯联邦, 85, Pobedа Str., Belgorod, Russia, 308015

Liliya Korokina

Belgorod State National Research University

Email: korokina@mail.ru
ORCID iD: 0000-0002-4115-1564

Candidate of Sciences (Medicine), Associate Professor, Associate Professor of the Department of Pharmacology

俄罗斯联邦, 85, Pobedа Str., Belgorod, Russia, 308015

Natalia Kartashkina

First Moscow State Medical University named after I. M. Sechenov (Sechenov University)

Email: kartashkuna_n_l@staff.sechenov.ru
ORCID iD: 0000-0003-4648-9027

Candidate of Sciences (Medicine), Associate Professor of the Department of histology, cytology and embryology

俄罗斯联邦, Bldg. 2, 8, Trubetskaya str., Moscow, Russia, 119991

Valeria Soldatova

Belgorod State National Research University

Email: lorsoldatova@gmail.com
ORCID iD: 0000-0001-6637-1654

Postgraduate student of the Department of Pharmacology

俄罗斯联邦, 85, Pobedа Str., Belgorod, Russia, 308015

Elena Kuzubova

Belgorod State National Research University

Email: 1015artek1015@mail.ru
ORCID iD: 0000-0003-2425-5027

Postgraduate student of the Department of Pharmacology and Clinical Pharmacology

俄罗斯联邦, 85, Pobedа Str., Belgorod, Russia, 308015

Alexandra Radchenko

Belgorod State National Research University

Email: sandrinkaradchenko@gmail.com
ORCID iD: 0000-0002-4554-2116

Postgraduate student of the Department of Pharmacology and Clinical Pharmacology

俄罗斯联邦, 85, Pobedа Str., Belgorod, Russia, 308015

Mikhail Pokrovskii

Belgorod State National Research University

Email: pokrovskii@bsu.edu.ru
ORCID iD: 0000-0002-1493-3376

Doctor of Sciences (Medicine), Professor, Head of the Department of Pharmacology and Clinical Pharmacology

俄罗斯联邦, 85, Pobedа Str., Belgorod, Russia, 308015

参考

  1. Jelkmann W. Erythropoietin after a century of research: younger than ever. Eur J Haematol. 2007 Mar;78(3):183–205. doi: 10.1111/j.1600-0609.2007.00818.x.
  2. Heikal L, Ghezzi P, Mengozzi M, Stelmaszczuk B, Feelisch M, Ferns GA. Erythropoietin and a nonerythropoietic peptide analog promote aortic endothelial cell repair under hypoxic conditions: role of nitric oxide. Hypoxia (Auckl). 2016 Aug 16;4:121–133. doi: 10.2147/HP.S104377.
  3. Zhang Y, Wang L, Dey S, Alnaeeli M, Suresh S, Rogers H, Teng R, Noguchi CT. Erythropoietin Action in Stress Response, Tissue Maintenance and Metabolism. International Journal of Molecular Sciences. 2014; 15(6):10296–10333. doi: 10.3390/ijms150610296.
  4. Wu H, Liu X, Jaenisch R, Lodish HF. Generation of committed erythroid BFU-E and CFU-E progenitors does not require erythropoietin or the erythropoietin receptor. Cell. 1995 Oct 6;83(1):59–67. doi: 10.1016/0092-8674(95)90234-1.
  5. Lin CS, Lim SK, D’Agati V, Costantini F. Differential effects of an erythropoietin receptor gene disruption on primitive and definitive erythropoiesis. Genes Dev. 1996 Jan 15;10(2):154–64. doi: 10.1101/gad.10.2.154.
  6. Ueba H, Shiomi M, Brines M, Yamin M, Kobayashi T, Ako J, Momomura S, Cerami A, Kawakami M. Suppression of coronary atherosclerosis by helix B surface Peptide, a nonerythropoietic, tissue-protective compound derived from erythropoietin. Mol Med. 2013 Jul 24;19(1):195–202. doi: 10.2119/molmed.2013.00037.
  7. Lu KY, Ching LC, Su KH, Yu YB, Kou YR, Hsiao SH, Huang YC, Chen CY, Cheng LC, Pan CC, Lee TS. Erythropoietin suppresses the formation of macrophage foam cells: role of liver X receptor alpha. Circulation. 2010 Apr 27;121(16):1828–37. doi: 10.1161/CIRCULATIONAHA.109.876839.
  8. Haine L, Yegen CH, Marchant D, Richalet JP, Boncoeur E, Voituron N. Cytoprotective effects of erythropoietin: What about the lung? Biomedicine & Pharmacotherapy. 2021;139:111547. doi: 10.1016/j.biopha.2021.111547.
  9. Brines M, Grasso G, Fiordaliso F, Sfacteria A, Ghezzi P, Fratelli M, Latini R, Xie QW, Smart J, Su-Rick CJ, Pobre E, Diaz D, Gomez D, Hand C, Coleman T, Cerami A. Erythropoietin mediates tissue protection through an erythropoietin and common beta-subunit heteroreceptor. Proc Natl Acad Sci USA. 2004 Oct 12;101(41):14907–12. doi: 10.1073/pnas.0406491101.
  10. Brines M, Grasso G, Fiordaliso F, Sfacteria A, Ghezzi P, Fratelli M, Latini R, Xie QW, Smart J, Su-Rick CJ, Pobre E, Diaz D, Gomez D, Hand C, Coleman T, Cerami A. Erythropoietin mediates tissue protection through an erythropoietin and common beta-subunit heteroreceptor. Proceedings of the National Academy of Sciences USA. 2001;101(41): 14907–12. doi: 10.1073/pnas.0406491101.
  11. Anagnostou A, Lee ES, Kessimian N, Levinson R, Steiner M. Erythropoietin has a mitogenic and positive chemotactic effect on endothelial cells. Proceedings of the National Academy of Sciences USA. 1990;87: 5978–82. doi: 10.1073/pnas.87.15.5978.
  12. Hou J, Wang S, Shang YC, Chong ZZ, Maiese K. Erythropoietin employs cell longevity pathways of SIRT1 to foster endothelial vascular integrity during oxidant stress. Curr Neurovasc Res. 2011 Aug 1;8(3):220–35. doi: 10.2174/156720211796558069.
  13. Beleslin-Čokić BB, Cokić VP, Wang L, Piknova B, Teng R, Schechter AN, Noguchi CT. Erythropoietin and hypoxia increase erythropoietin receptor and nitric oxide levels in lung microvascular endothelial cells. Cytokine. 2011 May;54(2):129–35. doi: 10.1016/j.cyto.2011.01.015.
  14. Kanagy NL, Perrine MF, Cheung DK, Walker BR. Erythropoietin administration in vivo increases vascular nitric oxide synthase expression. J Cardiovasc Pharmacol. 2003 Oct;42(4):527–33. doi: 10.1097/00005344-200310000-00011.
  15. Corwin HL, Gettinger A, Fabian TC, May A, Pearl RG, Heard S, An R, Bowers PJ, Burton P, Klausner MA, Corwin MJ; EPO Critical Care Trials Group. Efficacy and safety of epoetin alfa in critically ill patients. N Engl J Med. 2007 Sep 6;357(10):965–76. doi: 10.1056/NEJMoa071533.
  16. Brines M, Patel NS, Villa P, Brines C, Mennini T, De Paola M, Erbayraktar Z, Erbayraktar S, Sepodes B, Thiemermann C, Ghezzi P, Yamin M, Hand CC, Xie QW, Coleman T, Cerami A. Nonerythropoietic, tissue-protective peptides derived from the tertiary structure of erythropoietin. Proc Natl Acad Sci USA. 2008 Aug 5;105(31):10925–30. doi: 10.1073/pnas.0805594105.
  17. Erbayraktar Z, Erbayraktar S, Yilmaz O, Cerami A, Coleman T, Brines M. Nonerythropoietic tissue protective compounds are highly effective facilitators of wound healing. Molecular Medicine. 2009;15: 235–241. doi: 10.2119/molmed.2009.00051.
  18. Ueba H, Brines M, Yamin M, Umemoto T, Ako J, Momomura S, Cerami A, Kawakami M. Cardioprotection by a nonerythropoietic, tissue-protective peptide mimicking the 3D structure of erythropoietin. Proceedings of the National Academy of Sciences USA. 2010;107:14357–62. doi: 10.1073/pnas.1003019107.
  19. Ahmet I, Tae HJ, Juhaszova M, Riordon DR, Boheler KR, Sollott SJ, Brines M, Cerami A, Lakatta EG, Talan MI. A small nonerythropoietic helix B surface peptide based upon erythropoietin structure is cardioprotective against ischemic myocardial damage. Mol Med. 2011 Mar–Apr;17(3–4):194–200. doi: 10.2119/molmed.2010.00235.
  20. Puchenkova OA, Nadezhdin SV, Soldatov VO, Zhuchenko MA, Korshunova DS, Kubekina MV, Korshunov EN, Korokina LV, Golubinskaya PA, Kulikov AL, Gureev VV, Pokrovskiy VM, Patrakhanov EA, Lebedev PR, Denisyuk TA, Belyaeva VS, Movchan EA, Lepetukha EI, Pokrovskiy MV. Study of antiatherosclerotic and endothelioprotective activity of peptide agonists of EpoR/CD131 heteroreceptor. Pharmacy & Pharmacology. 2020;8(2):100–111. doi: 10.19163/2307-9266-2020-8-2-100-111.
  21. Stubbendorff M, Hua X, Deuse T, Ali Z, Reichenspurner H, Maegdefessel L, Robbins RC, Schrepfer S. Inducing myointimal hyperplasia versus atherosclerosis in mice: an introduction of two valid models. J Vis Exp. 2014 May 14;(87):51459. doi: 10.3791/51459.
  22. Tediashvili G, Wang D, Reichenspurner H, Deuse T, Schrepfer S. Balloon-based Injury to Induce Myointimal Hyperplasia in the Mouse Abdominal Aorta. J Vis Exp. 2018 Feb 7;(132):56477. doi: 10.3791/56477.
  23. Teng R, Calvert JW, Sibmooh N, Piknova B, Suzuki N, Sun J, Martinez K, Yamamoto M, Schechter AN, Lefer DJ, Noguchi CT. Acute erythropoietin cardioprotection is mediated by endothelial response. Basic Res Cardiol. 2011 May;106(3):343–54. doi: 10.1007/s00395-011-0158-z.
  24. Yasuda H, Iwata Y, Nakajima S, Furuichi K, Miyake T, Sakai N, Kitajima S, Toyama T, Shinozaki Y, Sagara A, Miyagawa T, Hara A, Shimizu M, Kamikawa Y, Sato K, Oshima M, Yoneda-Nakagawa S, Kaneko S, Wada T. Erythropoietin signal protected human umbilical vein endothelial cells from high glucose-induced injury. Nephrology (Carlton). 2019 Jul;24(7):767–774. doi: 10.1111/nep.13518.
  25. Beleslin-Cokic BB, Cokic VP, Yu X, Weksler BB, Schechter AN, Noguchi CT. Erythropoietin and hypoxia stimulate erythropoietin receptor and nitric oxide production by endothelial cells. Blood. 2004 Oct 1;104(7):2073–80. doi: 10.1182/blood-2004-02-0744.
  26. Korokin MV, Soldatov VO, Tietze AA, Golubev MV, Belykh AE, Kubekina MV, Puchenkova OA, Denisyuk TA, Gureyev VV, Pokrovskaya TG, Gudyrev OS, Zhuchenko MA, Zatolokina MA, Pokrovskiy MV. 11-amino acid peptide imitating the structure of erythropoietin α-helix b improves endothelial function, but stimulates thrombosis in rats. Pharmacy & Pharmacology. 2019;7(6):312–320. doi: 10.19163/2307-9266-2019-7-6-312-320.
  27. Shokrzadeh M, Etebari M, Ghassemi-Barghi N. An engineered non-erythropoietic erythropoietin-derived peptide, ARA290, attenuates doxorubicin induced genotoxicity and oxidative stress. Toxicol In Vitro. 2020 Aug;66:104864. doi: 10.1016/j.tiv.2020.104864.
  28. Belyaeva VS, Stepenko YuV, Lyubimov II, Kulikov AL, Tietze AA, Kochkarova IS, Martynova OV, Pokopeyko ON, Kru-pen’kina LA, Nagikh AS, Pokrovskiy VM, Patrakhanov EA, Belashova AV, Lebedev PR, Gureeva AV. Non-hematopoietic erythropoietin-derived peptides for atheroprotection and treatment of cardiovascular diseases. Research Results in Pharmacology. 2020;6(3): 75–86. doi: 10.3897/rrpharmacology.6.58891.
  29. Golubev IV, Gureev VV, Korokin MV, Zatolokina MA, Avdeeva EV, Gureeva AV, Rozhkov IS, Serdyuk EA, Soldatova VA Preclinical study of innovative peptides mimicking the tertiary structure of the α-helix B of erythropoietin. Research Results in Pharmacology. 2020;6(2):85–96. doi: 10.3897/rrpharmacology.6.55385.
  30. Golubev IV, Gureev VV, Korokina LV, Gudyrev OS, Pokrovskaia TG, Pokopeiko ON, Pokrovskii VM, Artyushkova EB, Korokin MV. The anti-aggregation activity of new 11-amino acid of erythropoietin derivate containing tripeptide motifs. Archivos venezolanos de farmacología y terapéutica. 2020;39(5):588–91. doi: 10.5281/zenodo.4264989.
  31. Warren JS, Zhao Y, Yung R, Desai A. Recombinant human erythropoietin suppresses endothelial cell apoptosis and reduces the ratio of Bax to Bcl-2 proteins in the aortas of apolipoprotein E-deficient mice. J Cardiovasc Pharmacol. 2011 Apr;57(4):424–33. doi: 10.1097/FJC.0b013e31820d92fd.
  32. Nairz M, Sonnweber T, Schroll A, Theurl I, Weiss G. The pleiotropic effects of erythropoietin in infection and inflammation. Microbes Infect. 2012 Mar;14(3):238–46. doi: 10.1016/j.micinf.2011.10.005.

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2. Figure 1 – Atherosclerotic plaque size in groups of wildtype and Polgmut/mut/Cdh5-CRE animals against the background of studied peptides use

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3. Figure 2 – Influence of test drugs on relative expression of apoptosis markers (2A) and Bax/Bcl-2 ratio (2B)

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4. Figure 3 – Influence of test compounds on inflammatory markers relative expression

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5. Figure 4 – Influence of test compounds on relative expression of related to angiogenesis factors

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6. Figure 5 – Results of a test with endothelial-dependent vasodilation in response to ACh on the isolated segments of thoracic aortas of the Polgmut/mut/Cdh5-CRE mice

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7. Figure 6 – Results of endothelial-independent vasodilation test in response to SN on isolated segments of thoracic aortas of Polgmut/mut/Cdh5-CRE mice

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版权所有 © Korokin M.B., Kubekina M.V., Deykin A.V., Antsiferov O.V., Pokrovskii V.M., Korokina L.V., Kartashkina N.L., Soldatova V.A., Kuzubova E.V., Radchenko A.I., Pokrovskii M.V., 2021

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