online ISSN 2415-3176
print ISSN 1609-6371
logoExperimental and Clinical Physiology and Biochemistry
  • 5 of 9
ECPB 2020, 89(1): 36–43
Research articles

The aminoguanidine effect on the content of cerebellar glial fibrillary acidic protein in experimental antiphospholipid syndrome with pregnancy


Antiphospholipid syndrome (APS) is an autoimmune disease that is characterized by arterial or venous thrombosis, thrombocytopenia. APS is one of the autoimmune causes of pregnancy miscarriages. According to the World Health Organization, the incidence of pregnancy miscarriages in the world is 15-25%. Habitual miscarriage is a multifactorial, genetically determined disease. In 50% of cases of habitual miscarriage the establishment of its cause is not possible. About 80% of unexplained reproductive losses are due to immunological disorders. The pathogenetic mechanisms of pregnancy loss mediated by antiphospholipid antibodies (aPL) are thrombosis, placental dysfunction and local inflammation. The nitric oxide system is one of the significant links in the mechanisms of APS development. The effect of selective inhibitor of aminoguanidine inducible NO synthase on the content of glial fibrillary acidic protein (GFAP) and the level of nitric oxide (NO) synthesis in the cerebellum in cases of APS in the pregnant BALB/c mice have been studied. The experimental animals were divided into 5 groups: the 1st – control; the 2nd – animals with experimental APS, the 3rd – animals with APS and administration of aminoguanidine (10 mg/kg). Aminoguanidine was administered intraperitoneally once a day, on a daily basis for 10 days before fertilization and for 17 days during pregnancy. After APS confirmation (on the 10th day), the females of all groups were mated with the males. The animals were removed from the experiment on the 18th day of pregnancy. Western blot analysis for GFAP in cerebellar samples of the control and experimental groups of mice was conducted as well as the densitometric analysis of immunoreactive zones. The content of NO in the samples was determined by the number of its stable metabolites of nitrite anions (NO2¯) and nitrate anions (NO3¯). During the studies, a 1.9 times increase in GFAP (total) and 12.9 times GFAP (49-37 kDa) in the cerebellum of the APS mice on the 18th day of pregnancy as compared to the control group has been established. The increase in GFAP content in the cerebellum of the BALB/c mice with APS is the evidence of the development of reactive astrogliosis. The increase in the content of stable metabolites of nitric oxide NO2 ¯ by 54% and NO3 ¯ by 65% in the cerebellum of mice with APS was found relative to the control group. With the administration of aminoguanidine in the cerebellum of the pregnant mice with APS, a decrease of GFAP (total) by 26%, GFAP (49-37 kDa) by 89%, NO2 ¯ by 31% and NO3 ¯ by 30% was observed in comparison with the indicators of the group of pregnant animals with APS. Therefore, aminoguanidine has a neuroprotective effect in antiphospholipid syndrome with underlying pregnancy which may be mediated not only by inhibiting the activity of inducible NO synthase but also through other cellular targets.

Recieved: 12.02.2020

Keywords: antiphospholipid syndrome, cerebellum, nitric oxide, glial fibrillary acidic protein, aminoguanidine

Full text: PDF (Ukr) 401K

  1. 1. Arachchillage DRJ, Laffan M. Pathogenesis and management of antiphospholipid syndrome. British Journal of Haematology 2017;178:181-95.
  2. 2. Fleetwood T, Cantello R, Comi C. Antiphospholipid Syndrome and the Neurologist: From Pathogenesis to Therapy. Front. Neurol. 2018;9:1001.
  3. 3. Rahman A. Antiphospholipid syndrome in pregnancy. Indian J Rheumatol 2016;11:117-21.
  4. 4. Alves JD, Mason LJ, Ames PRJ, Chen PP, Rauch J, Levine JS et al. Antiphospholipid antibodies are associated with enhanced oxidative stress, decreased plasma nitric oxide and paraoxonase activity in an experimental mouse model. Rheumatology. 2005;44:1238-44.
  5. 5. Ferreira EI, Serafim RAM. Nitric Oxide Synthase Inhibitors, Nitric Oxide Synthase - Simple Enzyme-Complex Roles, Seyed Soheil Saeedi Saravi, IntechOpen, 2017.
  6. 6. Atochin DN. Endothelial nitric oxide synthase transgenic models of endothelial dysfunction. Pflugers Arch. 2010;460(6):965-74.
  7. 7. Moore C., Tymvios C., Emerson M. Functional regulation of vascular and platelet activity during thrombosis by nitric oxide and endothelial nitric oxide synthase. Thromb Haemost. 2010;104(2):342-9.
  8. 8. Cella M, Farina MG, Dominguez AP, Girolamo RD, Ribeiro ML, Franchi AM. Dual effect of nitric oxide on uterine prostaglandin synthesis in a murine model of preterm labour. Br J Pharmacol. 2010;161(4):844-55.
  9. 9. Song Y, Zhang F, Ying C, Kumar KA, Zhou X. Inhibition of NF-κB activity by aminoguanidine alleviates neuroinflammation induced by hyperglycemia. Metab Brain Dis. 2017;32(5):1627-37.
  10. 10. Dingman A, Lee SY, Derugin N, Wendland MF, Vexler ZS. Aminoguanidine inhibits caspase-3 and calpain activation without affecting microglial activation following neonatal transient cerebral ischemia. Journal of Neurochemistry. 2006;
  11. 11. Tykhomyrov АA, Pavlova AS, Nedzvetsky VS. Glial Fibrillary Acidic Protein (GFAP): on the 45th Anniversary of Its Discovery. Neurophysiology. 2016;48:54-71.
  12. 12. Zaichenko GV, Laryanovskaya YB, Deyeva TV, Shevchenko OI, Starokozhko VY, Kudina OV et al. Morphological status of the uterus and placenta in experimental model of gestational antiphospholipid syndrome on mice. Ukrainian Medical Almanac 2011;14(4):136-41.
  13. 13. Lowry ОМ, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 1951;193(1):265-75.
  14. 14. Laemmli UK. Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4. Nature. 1970;227:680-5.
  15. 15. Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA. 1979;76(9):4350-4.
  16. 16. Pavlova O. S., Tykhomyrov A. A., Mejenskaya O. A., Stepanenko S. P., Chehivska L. I., Parkhomenko Yu. M. High thiamine dose restores levels of specific astroglial proteins in rat brain astrocytes affected by chronic ethanol consumption. Ukr. Biochem. J. 2019;91(4):41-9.
  17. 17. Green LC, Davie AW, Golawski J. Analisis of nitrate, nitrite and [15N] nitrate in biological fluids. Anal. biochem. 1982;126(1):131-8.
  18. 18. Kiselik IO, Lutsik MD, Shevchenko LJ. Features of determination of nitrates and nitrites in blood of patients with viral hepatitis and jaundice of other etiology. Lab. diagnostics. 2001;3:43-5.
  19. 19. Richa R, Yadawa AK, Chaturvedi CM. Hyperglycemia and high nitric oxide level induced oxidative stress in the brain and molecular alteration in the neurons and glial cells of laboratory mouse, Mus musculus. Neurochem Int. 2017;104:64-79.
  20. 20. Bellaver B, Souza DG, Souza DO, Quincozes-Santos A. Hippocampal astrocyte cultures from adult and aged rats reproduce changes in glial functionality observed in the aging brain. Mol Neurobiol. 2016;54:2969-285.
  21. 21. Svenungsson E, Andersson M, Brundin L, van Vollenhoven R, Khademi M, Tarkowski A, et al. Increased levels of proinflammatory cytokines and nitric oxide metabolites in neuropsychiatric lupus erythematosus. Ann Rheum Dis. 2001;60:372-9.
  22. 22. Reutov VP, Sorokina EG, Samosudova NV, Zakharchuk NV. Brain hemodynamics: glutamatergic system and nitric oxide cycle in the regulation of cerebral circulation. New concept. Pacific Medical Journal. 2017; 3: 37-45.
  23. 23. Bayır H, Kagan VE, Borisenko GG, Tyurina YY, Janesko KL, Vagni VA et al. Enhanced oxidative stress in iNOS-deficient mice after traumatic brain injury: support for a neuroprotective role of iNOS. Journal of Cerebral Blood Flow & Metabolism. 2005;25:673-84.
  24. 24. Genc H, Baysal B, Eren B, Yılmaz BD. The Protective Effect of Amino-guanidine, an Inducible Nitric Oxide Synthase Inhibitor, on Aluminium Sulphate Neuro-toxicity in the Rat (Wistar albino) Cerebellar Purkinje Cells: Stereological Study. Middle Black Sea Journal of Health Science. 2017;3(3):7-14
  25. 25. Anaeigoudari A, Soukhtanloo M, Reisi P, Beheshti F, Hosseini M. Inducible nitric oxide inhibitor aminoguanidine, ameliorates deleterious effects of lipopolysaccharide on memory and long term potentiation in rat. Life Sciences. 2016;158:22-30.
  26. 26. Danielisova V, Burda J, Nemethova M, Gottlieb M. Aminoguanidine Administration Ameliorates Hippocampal Damage After Middle Cerebral Artery Occlusion in Rat. Neurochem Res. 2011;36:476-86.
  27. 27. Tsuji M, Higuchi Y, Shiraishi K., Kume T, Akaike A, Hattori H. Protective Effect of Aminoguanidine on Hypoxic-Ischemic Brain Damage and Temporal Profile of Brain Nitric Oxide in Neonatal Rat. Pediatr Res. 2000;47:79.
  28. 28. Cash D, Beech JS, Rayne R, Bath PMW, Meldrum BS, Williams SCR. Neuroprotective effect of aminoguanidine on transient focal ischaemia in the rat brain. Brain Research. 2001;905(1- 2):91-103.

Програмування -