A study of the mechanisms of action of fertiwell in vivo

Мұқаба

Дәйексөз келтіру

Толық мәтін

Ашық рұқсат Ашық рұқсат
Рұқсат жабық Рұқсат берілді
Рұқсат жабық Рұқсат ақылы немесе тек жазылушылар үшін

Аннотация

Aim. To investigate the specific mechanisms of action of Fertiwell in a mouse model of D-galactose-induced aging of the reproductive system.

Materials and methods. C57BL/6J mice were randomized into four groups: intact mice (control group), a group of mice with artificial accelerated aging treated with D-galactose alone (Gal), D-galactose followed by Fertiwell (PP), and D-galactose followed by a combination of L-carnitine and acetyl-L-carnitine (LC). The artificial accelerated aging of reproductive system was induced by daily intraperitoneal administration of D-galactose at a dose of 100 mg/kg for 8 weeks. After the end of therapy in all groups, the characteristics of sperm, the level of serum testosterone, immunohistochemical parameters, and the expression of specific proteins were evaluated.

Results. Fertiwell had a pronounced therapeutic effect on testicular tissues and spermatozoa, restored testosterone levels to normal values, and, in addition, was more effective protector against oxidative stress in the reproductive system compared to L-carnitine and acetyl-L-carnitine, which are widely used in male infertility. Fertiwell at a dose of 1 mg/kg allowed to significantly increase the number of motile spermatozoa to 67.4±3.1%, which was comparable to indicators in the intact group. The introduction of the Fertiwell positively affected the activity of mitochondria, which was also expressed in an increase in sperm motility. In addition, Fertiwell restored the intracellular level of ROS to the values of the control group and reduced the number of TUNEL+ cells (with fragmented DNA) to the level of intact control. Thus, Fertiwell, containing testis polypeptides, has a complex effect on reproductive function, leading to a change in gene expression, an increase in protein synthesis, the prevention of DNA damage in the testicular tissue, and an increase in mitochondrial activity in testicular tissue and spermatozoa of the vas deferens, which leads to the subsequent improvement of testicular function.

Толық мәтін

Рұқсат жабық

Авторлар туралы

Yu. Khochenkova

N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia

Хат алмасуға жауапты Автор.
Email: julia_vet@bk.ru

junior researcher of the Laboratory of biomarkers and mechanisms of tumor angiogenesis

Ресей, Moscow

Yu. Machkova

N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia

Email: knowl@mail.ru

junior researcher of the Laboratory of biomarkers and mechanisms of tumor angiogenesis

Ресей, Moscow

D. Khochenkov

N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia; Tolyatti State University

Email: khochenkov@gmail.com

Ph.D. in Biology, Head of the Laboratory of biomarkers and mechanisms of tumor angiogenesis, professor at the Center of Medical Chemistry 

Ресей, Moscow; Tolyatti

T. Sidorova

N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia

Email: tatsig51@yahoo.com

Ph.D., leading researcher of Laboratory of biomarkers and mechanisms of tumor angiogenesis

Ресей, Moscow

E. Safarova

PeptidPRO LLC

Email: elsaf2308@bk.ru

Ph.D. in Biology, freelance R&D consultant

Ресей, Moscow

N. Bastrikova

PeptidPRO LLC

Email: n.bastrikova@peptidpro.com

Ph.D. in Biology, Medical Director

Ресей, Moscow

K. Korzhova

PeptidPRO LLC

Email: k.korzhova@peptidpro.com

Ph.D. in Biology, preclinical research manager

Ресей, Moscow

Әдебиет тізімі

  1. Babakhanzadeh E., Nazari M., Ghasemifar S., Khodadadian A. Some of the Factors Involved in Male Infertility: A Prospective Review. Int J Gen Med. 2020;13:29–41. doi: 10.2147/IJGM.S241099.
  2. Bisht S., Faiq M., Tolahunase M., Dada R. Oxidative stress and male infertility. Nat Rev Urol. 2017;14:470–485. doi: 10.1038/nrurol.2017.69.
  3. Desai N., Sabanegh E., Kim T., Agarwal A. Free radical theory of aging: implications in male infertility. Urology. 2010;75:14–19. doi: 10.1016/j.urology.2009.05.025.
  4. Belloc S., Hazout A., Zini A., et al. How to overcome male infertility after 40: Influence of paternal age on fertility. Maturitas. 2014;78:22–29. doi: 10.1016/j.maturitas.2014.02.011.
  5. Parameshwaran K., Irwin M.H., Steliou K., Pinkert C.A. D-Galactose Effectiveness in Modeling Aging and Therapeutic Antioxidant Treatment in Mice. Rejuvenation Res. 2010;13:729–735. doi: 10.1089/rej.2010.1020.
  6. Salman T.M., Olayaki L.A., Alagbonsi I.A., Oyewopo A.O. Spermatotoxic effects of galactose and possible mechanisms of action. Middle East Fertil Soc J. 2016;21:82–90. doi: 10.1016/j.mefs.2015.09.004.
  7. Liu W., Zhang L., Gao A. et al. Food-Derived High Arginine Peptides Promote Spermatogenesis Recovery in Busulfan Treated Mice. Front Cell Dev Biol. 2021;9. Available: https://www.frontiersin.org/articles/10.3389/fcell.2021.791471
  8. Amaral A., Castillo J., Ramalho-Santos J., Oliva R. The combined human sperm proteome: cellular pathways and implications for basic and clinical science. Hum Reprod Update. 2014;20:40–62. doi: 10.1093/humupd/dmt046.
  9. Bu T., Wang L., Wu X. et al. A laminin-based local regulatory network in the testis that supports spermatogenesis. Semin Cell Dev Biol. 2022;121:40–52. doi: 10.1016/j.semcdb.2021.03.025.
  10. Wu S., Yan M., Ge R., Cheng C.Y. Crosstalk between Sertoli and Germ Cells in Male Fertility. Trends Mol Med. 2020;26:215–231. doi: 10.1016/j.molmed.2019.09.006.
  11. Rizzetti D.A., Martinez C.S., Escobar A.G. et al. Egg white-derived peptides prevent male reproductive dysfunction induced by mercury in rats. Food Chem Toxicol Int J Publ Br Ind Biol Res Assoc. 2017;100:253–264. doi: 10.1016/j.fct.2016.12.038.
  12. Wu Y., Tian Q., Li L. et al. Inhibitory effect of antioxidant peptides derived from Pinctada fucata protein on ultraviolet-induced photoaging in mice. J Funct Foods. 2013;5:527–538. doi: 10.1016/j.jff.2013.01.016.
  13. EAU Guidelines on Sexual and Reproductive Health - Uroweb. In: Uroweb - European Association of Urology [Internet]. [cited 22 Aug 2022]. Available: https://uroweb.org/guidelines/sexual-and-reproductive-health
  14. Pushkar D.Yu., Kupriyanov Y.A., Gamidov S.I., Teteneva A.V., Spivak L.G., Shormanov I.S., Novikov A.I., Al-Shukri S.Kh., Bogdan E.N., Shchukin V.L., Boriskin A.G. Assessment of the safety and efficacy of medicinal product PPR-001 based on regulatory polypeptides of the testes. Urologiia. 2021;6:100–108. Russian (Пушкарь Д.Ю., Куприянов Ю.А., Берников А.Н., Гамидов С.И., Тетенева А.В., Спивак Л.Г., Шорманов И.С., Новиков А.И., Аль-Шукри С.Х., Богдан Е.Н., Щукин В.Л., Борискин А.Г. Оценка безопасности и эффективности лекарственного препарата на основе регуляторных полипептидов семенников PPR-001. Урология. 2021;6:100–108). doi: 10.18565/urology.2021.6.100-108.
  15. Agarwal A., Majzoub A., Parekh N., Henkel R. A Schematic Overview of the Current Status of Male Infertility Practice. World J Mens Health. 2020;38:308–322. doi: 10.5534/wjmh.190068.
  16. Liao C.-H., Chen B.-H., Chiang H.-S. et al. Optimizing a Male Reproductive Aging Mouse Model by d-Galactose Injection. Int J Mol Sci. 2016;17:98. doi: 10.3390/ijms17010098.
  17. Beumer T.L., Roepers-Gajadien H.L., Gademan I.S. et al. The role of the tumor suppressor p53 in spermatogenesis. Cell Death Differ. 1998;5:669–677. doi: 10.1038/sj.cdd.4400396.
  18. Liang M., Wen J., Dong Q. et al. Testicular hypofunction caused by activating p53 expression induced by reactive oxygen species in varicocele rats. Andrologia. 2015;47: 1175–1182. doi: 10.1111/and.12400.
  19. Lindsay J., Esposti M.D., Gilmore A.P. Bcl-2 proteins and mitochondria-specificity in membrane targeting for death. Biochim Biophys Acta. 2011;1813: 532–539. doi: 10.1016/j.bbamcr.2010.10.017.
  20. Eliveld J., van den Berg E.A., Chikhovskaya J.V. et al. Primary human testicular PDGFRα+ cells are multipotent and can be differentiated into cells with Leydig cell characteristics in vitro. Hum Reprod Oxf Engl. 2019;34:1621–1631. doi: 10.1093/humrep/dez131.
  21. Basciani S., Mariani S., Arizzi M. et al. Expression of Platelet-Derived Growth Factor-A (PDGF-A), PDGF-B, and PDGF Receptor-α and -β during Human Testicular Development and Disease. J Clin Endocrinol Metab. 2002;87:2310–2319. doi: 10.1210/jcem.87.5.8476.
  22. Nelson J.F., Latham K.R., Finch C.E. Plasma testosterone levels in C57BL/6J male mice: effects of age and disease. Acta Endocrinol (Copenh). 1975;80:744–752. doi: 10.1530/acta.0.0800744.

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2. Fig. 1. Research design

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3. Fig. 2. The number of motile spermatozoa on days TR1,10,20,30 after administration of the drug Fertivell, or Ac-L-carnitine with L-carnitine, or saline solution (*, **, ***, **** p<0.05 is a significant difference compared to the Gal group, #, ##, ###, #### — p<0.05 significant difference between PP group and LC group)

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4. Fig. 3. Mitochondrial activity of spermatozoa (Mitotracker Green staining). The blue arrow indicates the direction of increased staining and, consequently, an increase in the number of mitochondria, the red arrow indicates the difference in mitochondrial activity between the intact group (Ctrl group, blue) and the experimental groups PP, LC and Gal (the group is indicated on the left side of the figure)

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5. Fig. 4. Oxidative stress in the spermatozoa of the vas deferens and levels of ROS (reactive oxygen species) (CellRox DeepRed staining). The blue arrow indicates the direction of the increase in staining and, consequently, the increase in ROS levels, the red arrow indicates the difference in ROS levels between the intact group (Ctrl group, blue) and the experimental groups PP, LC and Gal (the group is indicated in the left part of the figure)

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6. Fig. 5. Determination of the level of DNA fragmentation by the TUNEL method on day T20 (arrow shows TUNEL+ cells, green). The cores are colored using DAPI (blue). Magnification x 100

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7. Fig. 6. Immunohistochemical staining of testes in mice on day TR20 (arrow shows positively stained cells). Expression of proteins. Magnification *200

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8. Fig. 7. Possible mechanisms of action of the drug Fertivell

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