Russian Journal of PhysiologyRussian Journal of PhysiologyРоссийский физиологический журнал им. И.М. Сеченова0869-81392658-655XThe Russian Academy of Sciences65173710.31857/S0869813924100127VQIPECEXPERIMENTAL ARTICLESЭКСПЕРИМЕНТАЛЬНЫЕ СТАТЬИResearch ArticleThe effect of ACTH/MSH N-terminal fragment analogs on the anxiety level, pain sensitivity and levels of neurotrophic factors BDNF and VEGF in primary neuronal cultures of ratsВлияние аналогов N-концевых фрагментов АКТГ/МСГ на уровень тревожности, болевую чувствительность и уровни нейротрофических факторов BDNF и VEGF в первичных культурах нейронов мозга крысыGlazovaN. Yu.ГлазоваН. Ю.
Russian Federation
nglevitskaya@gmail.comManchenkoD. М.МанченкоД. М.
Russian Federation
nglevitskaya@gmail.comSebentsovaЕ. А.СебенцоваЕ. А.
Russian Federation
nglevitskaya@gmail.comAndreevaL. А.АндрееваЛ. А.
Russian Federation
nglevitskaya@gmail.comGrivennikovI. А.ГривенниковИ. А.
Russian Federation
nglevitskaya@gmail.comDolotovО. V.ДолотовО. В.
Russian Federation
nglevitskaya@gmail.comMyasoedovN. F.МясоедовН. Ф.
Russian Federation
nglevitskaya@gmail.comLevitskayaN. G.ЛевицкаяН. Г.
Russian Federation
nglevitskaya@gmail.comLomonosov Moscow State UniversityМосковский государственный университет имени М. В. ЛомоносоваNational Research Centre “Kurchatov Institute”Национальный исследовательский центр “Курчатовский институт”ru151020242024110101752176602022025Received:02022025Published:15102024Copyright ©; 2024, Russian Academy of SciencesCopyright ©; 2024, Российская академия наук2024Russian Academy of SciencesРоссийская академия наукhttps://journals.eco-vector.com/0869-8139/article/view/651737

ACTH/MSH-like peptides (melanocortins) have a wide range of neurotropic effects, including effects on learning and memory processes, neuroprotection, emotional state and pain sensitivity. Present work is aimed to compare the effects of peptides, the structure of which includes a natural fragment of ACTH and a stabilizing tripeptide PGP. The peptides ACTH4-7PGP (Semax), ACTH6-9PGP и ACTH7-10PGP were used in the work. The effects of these peptides on the exploratory behavior, anxiety level and pain sensitivity of white rats, as well as on the protein levels of the neurotrophic factors BDNF (brain derived neurotrophic factor) and VEGF (vascular endothelial growth factor) in primary neuron cultures were studied. A comparative study of the effects of analogs of different ACTH/MSH fragments revealed both similarities and differences in their neurotropic activity. The peptides structure of which includes a sequence of ACTH4-7 or ACTH6-9 have nootropic, anxiolytic and analgesic activity, and also cause an increase in VEGF levels in the culture of hippocampal neurons. The peptide containing the ACTH7-10 sequence in the structure exhibits anxiolytic activity, increases exploratory behavior, does not affect pain sensitivity and has a stimulating effect on BDNF and VEGF levels in neuronal cultures. The data obtained indicate that different parts of the N-terminal region of the ACTH molecule are responsible for the manifestation of certain neurotropic effects of melanocortins. The results of the study can be used in the development of therapeutics based on natural melanocortins.

АКТГ/МСГ-подобные пептиды (меланокортины) обладают широким спектром нейротропных эффектов, в том числе влияют на процессы обучения и памяти, нейропротекцию, эмоциональный статус и болевую чувствительность. Данная работа посвящена сравнению нейротропных эффектов пептидов, структура которых включает в себя природный фрагмент АКТГ и стабилизирующий трипептид PGP. В работе были использованы пептиды – АКТГ4-7PGP (семакс), АКТГ6-9PGP и АКТГ7-10PGP. Изучалось влияние этих пептидов на исследовательское поведение, уровень тревожности и болевую чувствительность белых крыс, а также на уровни нейротрофических факторов BDNF (brain derived neurotrophic factor) и VEGF (vascular endothelial growth factor) в первичных культурах нейронов мозга крысы. Сравнительное исследование эффектов аналогов различных фрагментов АКТГ/МСГ позволило выявить как сходство, так и различия их нейротропной активности. Пептиды, в структуре которых присутствует фрагмент АКТГ4-7 или АКТГ6-9, обладают ноотропной, анксиолитической и анальгетической активностью, а также вызывают повышение уровней VEGF в культуре нейронов гиппокампа. Пептид, содержащий в структуре последовательность АКТГ7-10, проявляет анксиолитическую активность, увеличивает исследовательское поведение, не влияет на болевую чувствительность и оказывает стимулирующее влияние на уровни BDNF и VEGF в нейрональных культурах. Полученные данные свидетельствуют о том, что разные участки N-концевой области молекулы АКТГ ответственны за проявление определенных нейротропных эффектов меланокортинов. Результаты исследования могут быть использованы при разработке лекарственных препаратов на основе природных меланокортинов.

melanocortinsACTH/MSH fragmentssynthetic analogsanxietypain sensitivityneurotrophic factorsratмеланокортиныфрагменты АКТГсинтетические аналогитревожностьболевая чувствительностьнейротрофические факторыкрысыПравительство Российской ФедерацииGovernment of the Russian Federation12586п-П8Feng W, Zhou Q, Chen X, Dai A, Cai X, Liu X, Zhao F, Chen Y, Ye C, Xu Y, Cong Z, Li H, Lin S, Yang D, Wang MW (2023) Structural insights into ligand recognition and subtype selectivity of the human melanocortin-3 and melanocortin-5 receptors. Cell Discov 9: 81. https://doi.org/10.1038/s41421-023-00586-4Bertolini A, Tacchi R, Vergoni AV (2009) Brain effects of melanocortins. Pharmacol Res 59: 13–47. https://doi.org/10.1016/j.phrs.2008.10.005Akimov MG, Fomina-Ageeva EV, Dudina PV, Andreeva LA, Myasoyedov NF, Bezuglov VV (2021) ACTH(6–9)PGP peptide protects SH-SY5Y cells from H2O2, tert-butyl Hydroperoxide, and cyanide cytotoxicity via stimulation of proliferation and induction of prosurvival-related genes. Molecules 26: 1878. https://doi.org/10.3390/molecules26071878Gebrie A (2023) The melanocortin receptor signaling system and its role in neuroprotection against neurodegeneration: Therapeutic insights. Ann N Y Acad Sci 1527: 30–41. https://doi.org/10.1111/nyas.15048Micioni Di Bonaventura E, Botticelli L, Del Bello F, Giorgioni G, Piergentili A, Quaglia W, Romano A, Gaetani S, Micioni Di Bonaventura MV, Cifani C (2022) Investigating the role of the central melanocortin system in stress and stress-related disorders. Pharmacol Res 185: 106521. https://doi.org/10.1016/j.phrs.2022.106521Markov DD, Dolotov OV, Grivennikov IA (2023) The Melanocortin System: A Promising Target for the Development of New Antidepressant Drugs. Int J Mol Sci 24: 6664. https://doi.org/10.3390/ijms24076664Ericson MD, Lensing CJ, Fleming KA, Schlasner KN, Doering SR, Haskell-Luevano C (2017) Bench-top to clinical therapies: A review of melanocortin ligands from 1954 to 2016 Biochim Biophys Acta Mol Basis Dis 1863: 2414–2435. https://doi.org/10.1016/j. bbadis.2017.03.020Mowlazadeh Haghighi S, Zhou Y, Dai J, Sawyer JR, Hruby VJ, Cai M (2018) Replacement of Arg with Nle and modified D-Phe in the core sequence of MSHs, Ac-His-D-Phe-Arg-Trp-NH2, leads to hMC1R selectivity and pigmentation. Eur J Med Chem 151: 815–823. https://doi.org/10.1016/j.ejmech.2018.04.021Todorovic A, Ericson MD, Palusak RD, Sorensen NB, Wood MS, Xiang Z, Haskell-Luevano C (2016) Comparative Functional Alanine Positional Scanning of the α-Melanocyte Stimulating Hormone and NDP-Melanocyte Stimulating Hormone Demonstrates Differential Structure-Activity Relationships at the Mouse Melanocortin Receptors. ACS Chem Neurosci 7: 984–994. https://doi.org/10.1021/acschemneuro.6b00098Strand FL (2000) David and Goliath – the slingshot that started the neuropeptide revolution. Eur J Pharmacol 405: 3–12. https://doi.org/10.1016/s0014-2999(00)00536-7Ivanova DM, Levitskaya NG, Andreeva LA, Kamenskii AA, Myasoedov NF (2007) Comparative study of analgesic potency of ACTH4-10 fragment and its analog semax. Bull Exp Biol Med 143: 5–8. https://doi.org/10.1007/s10517-007-0002-5Catania A (2008) Neuroprotective actions of melanocortins: a therapeutic opportunity. Trends Neurosci 31: 353–360. https://doi.org/10.1016/j.tins.2008.04.002Wolterink G, van Ree JM (1989) Behavioral and neurotrophic activity of ACTH-(7-16)NH2. Life Sci 45: 703–710. https://doi.org/10.1016/0024-3205(89)90089-1De Wied D (1999) Behavioral pharmacology of neuropeptides related to melanocortins and the neurohypophyseal hormones. Eur J Pharmacol 375: 1–11. https://doi.org/10.1016/s0014-2999(99)00339-8Smith JS, Lefkowitz RJ, Rajagopal S (2018) Biased signalling: from simple switches to allosteric microprocessors. Nat Rev Drug Discov 17: 243–260. https://doi.org/10.1038/nrd.2017.229Vyunova TV, Andreeva LA, Shevchenko KV, Glazova NY, Sebentsova EA, Levitskaya NG, Myasoedov NF (2023) Synthetic corticotropins and the GABA receptor system: Direct and delayed effects. Chem Biol & Drug Design 101: 1393–1405. https://doi.org/10.1111/cbdd.14221Singh A, Haslach EM, Haskell-Luevano C (2010) Structure-activity relationships (SAR) of melanocortin and agouti-related (AGRP) peptides. Adv Exp Med Biol 681: 1–18. https://doi.org/10.1007/978-1-4419-6354-3_1Ashmarin IP, Samonina GE, Lyapina LA, Kamenskii AA, Levitskaya NG, Grivennikov IA, Dolotov OV, Andreeva LA, Myasoedov NF (2005) Natural and hybrid (“chimeric”) stable regulatory glyproline peptides. Pathophysiology 11: 179–185. https://doi.org/10.1016/j. pathophys.2004.10.001Kolomin T, Shadrina M, Slominsky P, Limborska S, Myasoedov N (2013) A new generation of drugs: Synthetic peptides based on natural regulatory peptides. Neurosci and Med 4: 223–252. https://doi.org/10.4236/nm.2013.44035Levitskaya NG, Glazova NYu, Sebentsova EA, Manchenko DM, Vilensky DA, Andreeva LA, Kamensky AA, Myasoedov NF (2008) Investigation of the Spectrum of Physiological Activities of the Heptapeptide Semax, an ACTH 4–10 Analogue. Neurochem J 2: 95–101. https://doi.org/10.1007/s11710-008-1018-0Levitskaya NG, Vilenskii DA, Sebentsova EA, Andreeva LA, Kamensky AA, Myasoedov NF (2010) Influence of semax on the emotional state of white rats in the norm and against the background of cholecystokinin-tetrapeptide action. Biol Bull 37: 186–192. https://doi.org/10.1134/S1062359010020147Ivanova DM, Vilenskii DA, Levitskaya NG, Andreeva LA, Alfeeva LYu, Kamenskii AA, Myasoedov NF (2006) Study of the relationship between analgesic activity and structure of synthetic melanocortin analogs. Biol Bull 33: 162–166. https://doi.org/10.1134/S1062359006020105Shadrina MI, Dolotov OV, Grivennikov IA, Slominsky PA, Andreeva LA, Inozemtseva LS, Limborska SA, Myasoedov NF (2001) Rapid induction of neurotrophin mRNAs in rat glial cell cultures by Semax, an adrenocorticotropic hormone analog. Neurosci Lett 308: 115–118. https://doi.org/10.1016/s0304-3940(01)01994-2Dolotov OV, Karpenko EA, Inozemtseva LS, Seredenina TS, Levitskaya NG, Rozyczka J, Dubynina EV, Novosadova EV, Andreeva LA, Alfeeva LY, Kamensky AA, Grivennikov IA, Myasoedov NF, Engele J (2006) Semax, an analog of ACTH(4-10) with cognitive effects, regulates BDNF and trkB expression in the rat hippocampus. Brain Res 1117: 54–60. https://doi.org/10.1016/j.brainres.2006.07.108Левицкая НГ, Глазова НЮ, Себенцова ЕА, Манченко ДМ, Андреева ЛА, Каменский АА, Мясоедов НФ (2019) Ноотропные и анксиолитические эффекты гептапептида АКТГ6-9Pro-Gly-Pro. Рос физиол журн им ИМ Сеченова 105: 761–770. [Levitskaya NG, Glazova NY, Sebentsova EA, Manchenko DM, Andreeva LA, Kamensky AA, Myasoedov NF (2019) Nootropic and anxiolytic effects of heptapeptide ACTH6-9Pro-Gly-Pro. Russ J Physiol 105: 761–770. (In Russ)]. https://doi.org/10.1134/S0869813919060049Filippenkov IB, Stavchansky VV, Glazova NY, Sebentsova EA, Remizova JA, Valieva LV, Levitskaya NG, Myasoedov NF, Limborska SA, Dergunova LV (2021) Antistress action of melanocortin derivatives associated with correction of gene expression patterns in the hippocampus of male rats following acute stress. Int J Mol Sci 22: 10054. https://doi.org/10.3390/ijms221810054Vorvul AO, Bobyntsev II, Medvedeva OA, Mukhina AY, Svishcheva MV, Azarova IE, Andreeva LA, Myasoedov NF (2022) ACTH(6-9)-Pro-Gly-Pro ameliorates anxietylike and depressive-like behaviour and gut mucosal microbiota composition in rats under conditions of chronic restraint stress. Neuropeptides 93: 102247. https://doi.org/10.1016/j. npep.2022.102247Glazova NYu, Atanov MS, Pyzgareva AV, Andreeva LA, Manchenko DM, Markov DD, Inozemtseva LS, Dolotov OV, Levitskaya NG, Kamensky AA, Grivennikov IA, Myasoedov NF (2011) Neurotropic Activity of ACTH7–10PGP, an Analog of an ACTH Fragment. Dokl Biol Sci 440: 270–274. https://doi.org/10.1134/S0012496611050140Licht T, Goshen I, Avital A, Kreisel T, Zubedat S, Eavri R, Segal M, Yirmiya R, Keshet E (2011) Reversible modulations of neuronal plasticity by VEGF. Proc Natl Acad Sci U S A 108: 5081–5086. https://doi.org/10.1073/pnas.1007640108De Rossi P, Harde E, Dupuis JP, Martin L, Chounlamountri N, Bardin M, Watrin C, Benetollo C, Pernet-Gallay K, Luhmann HJ, Honnorat J, Malleret G, Groc L, Acker-Palmer A, Salin PA, Meissirel C (2016) A critical role for VEGF and VEGFR2 in NMDA receptor synaptic function and fear-related behavior. Mol Psychiatry 21: 1768–1780. https://doi.org/10.1038/mp.2015.195Dubynina EV, Inozemtseva LS, Markov DD, Yatsenko KA, Dolotov OV, Grivennikov IA (2009) Alpha-melanocyte-stimulating hormone increases the expression of vascular endothelial growth factor in rat hippocampal astrocytes in vitro. Neurochem J 3: 267–271. https://doi.org/10.1134/S1819712409040059Bottenstein JE, Sato GH (1979) Growth of a rat neuroblastoma cell line in serum-free supplemented medium. Proc Natl Acad Sci U S A 76: 514–517. https://doi.org/10.1073/pnas.76.1.514Pollock GS, Vernon E, Forbes ME, Yan Q, Ma YT, Hsieh T, Robichon R, Frost DO, Johnson JE (2001) Effects of early visual experience and diurnal rhythms on BDNF mRNA and protein levels in the visual system, hippocampus, and cerebellum. J Neurosci 21: 3923–3931. https://doi.org/10.1523/JNEUROSCI.21-11-03923.2001Manchenko DM, Glazova NY, Levitskaya N, Andreeva LA, Kamenskii AA, Myasoedov NF (2012) The Nootropic and Analgesic Effects of Semax Given via Different Routes. Neurosci Behav Physiol 42: 264–270. doi:10.1007/s11055-012-9562-6Pettersen VL, Zapata-Sudo G, Raimundo JM, Trachez MM, Sudo RT (2009) The synergistic interaction between morphine and maprotiline after intrathecal injection in rats. Anesth Analg 109: 1312–1327. https://doi.org/10.1213/ane.0b013e3181b16ff5Pereira LO, da Cunha IC, Neto JM, Paschoalini MA, Faria MS (2005) The gradient of luminosity between open/enclosed arms, and not the absolute level of Lux, predicts the behaviour of rats in the plus maze. Behav Brain Res 159: 55–61. https://doi.org/10.1016/j.bbr.2004.10.002Violle N, Balandras F, Le Roux Y, Desor D, Schroeder H (2009) Variations in illumination, closed wall transparency and/or extramaze space influence both baseline anxiety and response to diazepam in the rat elevated plus-maze. Behav Brain Res 203: 35–42. https://doi.org/10.1016/j.bbr.2009.04.015Padovan CM, Guimarães FS (2000) Restraint-induced hypoactivity in an elevated plus-maze. Braz J Med Biol Res 33: 79–83. https://doi.org/10.1590/s0100-879x2000000100011Walker JM, Berntson GG, Sandman CA, Kastin AG, Akil H (1981) Induction of Analgesia by Central Administration of ORG 2766, An Analog of ACTH 4-9. Eur J Pharmacol 69: 71–79. https://doi.org/10.1016/0014-2999(81)90603-8Huber R, Tononi G, Cirelli C (2007) Exploratory behavior, cortical BDNF expression, and sleep homeostasis. Sleep 30: 129–139. https://doi.org/10.1093/sleep/30.2.129Zhu SW, Codita A, Bogdanovic N, Hjerling-Leffler J, Ernfors P, Winblad B, Dickins DW, Mohammed AH (2009) Influence of environmental manipulation on exploratory behaviour in male BDNF knockout mice. Behav Brain Res 197: 339–346. https://doi.org/10.1016/j.bbr.2008.09.032Dayi A, Cetin F, Sisman AR, Aksu I, Tas A, Gönenc S, Uysal N (2015) The effects of oxytocin on cognitive defect caused by chronic restraint stress applied to adolescent rats and on hippocampal VEGF and BDNF levels. Med Sci Monit 21: 69–75. https://doi.org/10.12659/MSM.893159Nicoletti JN, Lenzer J, Salerni EA, Shah SK, Elkady A, Khalid S, Quinteros D, Rotella F, Betancourth D, Croll SD (2010) Vascular endothelial growth factor attenuates status epilepticus-induced behavioral impairments in rats. Epilepsy Behav 19: 272–277. https://doi.org/10.1016/j.yebeh.2010.07.011