Betaine Attenuates The Expression of Vasoactive Mediators and Histological Alterations Associated with Ovarian Hyperstimulation Syndrome in Rats

Document Type : Original Article


1 Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran

2 Department of Pharmacology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran

3 Department of Basic Sciences, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran

4 Department of Biochemistry and Molecular Biology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran

5 Stem Cells and Transgenic Technology Research Center, Shahid Chamran University of Ahvaz, Ahvaz, Iran


Objective: Ovarian hyperstimulation syndrome (OHSS) is one female reproductive disorder that can occur after
administration of injectable hormonal drugs to stimulate ovulation. Betaine (BET) is an intracellular biomolecule with
anti-inflammatory and tissue protective effects. There is no information about its effects in an experimental model of
OHSS. The current study aims to investigate the possible effects of BET on abnormal expressions of vasoconstrictor
proteins and ovarian histological changes in an experimental OHSS rat model.
Materials and Methods: In this experimental study, 30 adult female rats (two months old) were randomly divided into
six groups (n=5 per group): i. Control, ii. OHSS [10 IU sc equine chorionic gonadotropin (eCG) for 4 days followed by
30 IU sc human chorionic gonadotropin (hCG) on the fifth day], iii. OHSS+BET (200 mg/kg/day, orally for seven days),
iv. OHSS+Cabergoline (CAB, 100 mg/kg/day, orally for six days), v. BET, and vi. CAB. Expression levels of vascular
endothelial growth factor (VEGF), cyclooxygenase-2 (COX-2), and blood levels of oestradiol (E2) and progesterone
(P4) were measured at the end of the experiment. The ovaries were studied for histomorphological changes.
Results: Induction of OHSS altered tissue histology, including an increase in the number of corpora lutea and atretic
follicles, and decreased the number of follicular reserves. In this group, we observed increased expressions of the
VEGF and COX-2 proteins, and increased serum E2 and P4 levels. Administration of CAB and BET significantly
attenuated all molecular and histological alterations observed in the OHSS animals.
Conclusion: Our findings, for first time, indicate the beneficial effects of BET to reduce OHSS complications in patients
by reducing the expressions of vasoactive proteins and improving changes to the ovarian tissues. The findings are
similar to CAB and can be a new avenue for future research on BET.


  1. Eurell JA, Frappier BL. Dellmann’s textbook of veterinary histology. 6th edition. John Wiley &Sons; Wiley-Blackwell: 2013.
  2. Shpakov AO. Improvement effect of metformin on female and male reproduction in endocrine pathologies and its mechanisms. Pharmaceuticals (Basel). 2021; 14(1): 42.
  3. Petrenko AP, Castelo-Branco C, Marshalov DV, Kuligin AV, Shifman EM, Nesnova ES. Assessing the usefulness of severity markers in women with ovarian hyperstimulation syndrome. Reprod Sci. 2021; 28(4): 1041-1048.
  4. Tso LO, Leis L, Glina CG, Busso CE, Romano RS, Busso NE, et al. Does the controlled ovarian stimulation increase the weight of women undergoing IVF treatment? Eur J Obstet Gynecol Reprod Biol. 2021; 263: 205-209.
  5. Li Y, Fang L, Zhang R, Wang S, Li Y, Yan Y, et al. Melatonin stimulates VEGF expression in human granulosa-lutein cells: a potential mechanism for the pathogenesis of ovarian hyperstimulation syndrome. Mol Cell Endocrinol. 2020; 518: 110981.
  6. Vidal A, Wachter C, Kohl Schwartz A, Dhakal C. A rare presentation of isolated right-sided pleural effusion in the context of ovarian hyperstimulation syndrome: a case report. Case Rep Womens Health. 2021; 32: e00347.
  7. Kasap E, Turan GA, Eskicioğlu F, Cengiz H, Gur EB, Sivrikoz ON, et al. Comparison between resveratrol and cabergoline in preventing ovarian hyperstimulation syndrome in a rat model. Gynecol Endocrinol. 2016; 32(8): 634-640.
  8. Ferrero H, García-Pascual CM, Pellicer N, Simón C, Pellicer A, Gómez R. Dopamine agonist inhibits vascular endothelial growth factor protein production and secretion in granulosa cells. Reprod Biol Endocrinol. 2015; 13: 104.
  9. Hulde N, Rogenhofer N, Brettner F, Eckert NC, Fetz I, Buchheim JI, et al. Effects of controlled ovarian stimulation on vascular barrier and endothelial glycocalyx: a pilot study. J Assist Reprod Genet. 2021; 38(9): 2273-2282.
  10. Mohamed MO, Adam EA, Abdlla BM, Abdelghani S, Eltayeb LB. Vascular endothelial growth factor (VEGF) as biological markers expressed in ovarian cancer using immuno-histochemical technique. J Med Sci. 2021; 25(112): 1311-1137.
  11. Fouda UM, Elshaer HS, Youssef GG, Hanafy A, Mehrem WM, Youssef MA, et al. Cabergoline versus calcium infusion in the prevention of ovarian hyperstimulation syndrome: a randomised controlled study. J Obstet Gynaecol. 2022; 42(1): 122-126.
  12. Tang H, Mourad SM, Wang A, Zhai SD, Hart RJ. Dopamine agonists for preventing ovarian hyperstimulation syndrome. Cochrane Database Syst Rev. 2021; 4(4): CD008605.
  13. Liu L, Wang X, Meng T, Jyu J, Lyu F, Zhang X. The influence of cabergoline and coasting in prevention of the ovarian hyperstimulation syndrome in patients undergoing IVF/ICSI-ET treatment: a systematic review and meta-analysis. Adv Reprod Sci. 2020; 8(2): 143.
  14. Şanlı C, Atılgan R, Kuloğlu T, Pala Ş, Aydın Türk B, Keser HB, et al. Transient receptor potential melastatin 2 ion channel activity in ovarian hyperstimulation syndrome physiopathology. Turk J Med Sci. 2021; 51(2): 787-795.
  15. Rubenfeld ES, Dahan MH. Does the timing of cabergoline administration impact rates of ovarian hyperstimulation syndrome? Obstet Gynecol Sci. 2021; 64(4): 345-352.
  16. Oishi S, Mekaru K, Nakamura R, Miyagi M, Akamine K, Heshiki C, et al. Two cases of polycystic ovary syndrome with onset of severe ovarian hyperstimulation syndrome following controlled ovarian stimulation with aromatase inhibitors for fertility preservation before breast cancer treatment. Taiwan J Obstet Gynecol. 2021; 60(5): 931-934.
  17. Samie KA, Tabandeh MR, Afrough M. Betaine ameliorates impaired steroidogenesis and apoptosis in mice granulosa cells induced by high glucose concentration. Syst Biol Reprod Med. 2020; 66(6): 400-409.
  18. Zhao G, He F, Wu C, Li P, Li N, Deng J, et al. Betaine in inflammation: mechanistic aspects and applications. Front Immunol. 2018; 9: 1070.
  19. Engin-Ustun Y, Yılmaz S, Timur H, Ustun Y, Moraloglu O, Deveer R, et al. Comparison of bevacizumab and cabergoline in the treatment of ovarian hyperstimulation syndrome in a rat model. Gynecol Endocrinol. 2013; 29(9): 851-854.
  20. Fatemi I, Khalili H, Mehrzadi S, Basir Z, Malayeri A, Goudarzi M. Mechanisms involved in the possible protective effect of chrysin against sodium arsenite-induced liver toxicity in rats. Life Sci. 2021; 267: 118965.
  21. Darabi Z, Basir Z, Tabandeh MR, Ghotbeddin Z. Coenzyme Q10 improves ovarian histology and attenuates the expression of angiogenesis- associated proteins in the ovary of rats with experimental hyperstimulation syndrome. Iran J Basic Med Sci. 2022; 25(8): 989-996.
  22. Shields R, Vollenhoven B, Ahuja K, Talmor A. Ovarian hyperstimulation syndrome: a case control study investigating risk factors. Aust N Z J Obstet Gynaecol. 2016; 56(6): 624-627.
  23. Barua S, Hng TM, Smith H, Bradford J, McLean M. Ovulatory disorders are an independent risk factor for pregnancy complications in women receiving assisted reproduction treatments. Aust N Z J Obstet Gynaecol. 2017; 57(3): 286-293.
  24. Zheng X, Guo W, Zeng L, Zheng D, Yang S, Xu Y, et al. In vitro maturation without gonadotropins versus in vitro fertilization with hyperstimulation in women with polycystic ovary syndrome: a noninferiority randomized controlled trial. Hum Reprod. 2022; 37(2): 242-253.
  25. Yanagihara Y, Tanaka A, Nagayoshi M, Tanaka I, Shinohara R, Fukushima F, et al. A modified GnRH antagonist method in combination with letrozole, cabergoline, and GnRH antagonist for PCOS: Safe and effective ovarian stimulation to treat PCOS and prevent OHSS. Reprod Med Biol. 2021; 21(1): e12429.
  26. Višnová H, Papaleo E, Martin FS, Koziol K, Klein BM, Mannaerts B. Clinical outcomes of potential high responders after individualized FSH dosing based on anti-Müllerian hormone and body weight. Reprod Biomed Online. 2021; 43(6): 1019-1026.
  27. Dirik D, Kömüro-lu AU. The effect of infliximab on oxidative stress in ovarian tissue of the rat with ovarian hyperstimulation syndrome. East J Med. 2021; 26(3): 475-480.
  28. Soares SR. Etiology of OHSS and use of dopamine agonists. Fertil Steril. 2012; 97(3): 517-522.
  29. Hortu I, Karadadas E, Ozceltik G, Tavmergen E, Tavmergen Goker EN, Yigitturk G, et al. Oxytocin and cabergoline alleviate ovarian hyperstimulation syndrome (OHSS) by suppressing vascular endothelial growth factor (VEGF) in an experimental model. Arch Gynecol Obstet. 2021; 303(4): 1099-1108.
  30. Pala Ş, Atilgan R, Ozkan ZS, Kavak SB, Ilhan N, Akpolat N, et al. Effect of varying doses of tamoxifen on ovarian histopathology, serum VEGF, and endothelin 1 levels in ovarian hyperstimulation syndrome: an experimental study. Drug Des Devel Ther. 2015; 9: 1761-1766.
  31. Karabulut S, Korkmaz O, Erdem Altun C, Keskin I. A histopathological evaluation of ovarian hyperstimulation syndrome on reproductive and vital organs and the role of the VEGF-PKA pathway in a mouse model. Cells Tissues Organs. 2021; 210(3): 218-238.
  32. Zimmerman L, Willson S, Setton R, Schattman G. Ovarian stimulation for fertility preservation (different protocols). In: Grynberg M, Patrizio P, editors. Female and male fertility preservation. Switzerland: Springer, Cham; 2022; 119-129.
  33. Hong YH, Kim SK, Lee JR, Jee BC, Suh CS. Clinical efficacy of dual trigger with human chorionic gonadotropin and a gonadotropin- releasing hormone agonist for women undergoing fertility preservation. Reprod Med Biol. 2022; 21(1): e12440.
  34. Hafez EM, Gowayed SM, Nehela Y, Sakran RM, Rady AMS, Awadalla A, et al. Incorporated biochar-based soil amendment and exogenous glycine betaine foliar application ameliorate rice (Oryza sativa L.) tolerance and resilience to osmotic stress. Plants (Basel). 2021; 10(9): 1930.
  35. Egbuniwe IC, Uchendu CN, Obidike IR. Ameliorative effects of betaine and ascorbic acid on endocrine and erythrocytic parameters of sexually-maturing female Japanese quails during the dry season. J Therm Biol. 2021; 96: 102812.
  36. Taghvaei S, Sabouni F, Minuchehr Z, Taghvaei A. Identification of novel anti-cancer agents, applying in silico method for SENP1 protease inhibition. J Biomol Struct Dyn. 2022; 40(14): 6228-6242.
  37. Wu Y, Jennings NB, Sun Y, Dasari SK, Bayraktar E, Corvigno S, et al. Targeting CCR2+ macrophages with BET inhibitor overcomes adaptive resistance to anti-VEGF therapy in ovarian cancer. J Cancer Res Clin Oncol. 2022; 148(4): 803-821.
  38. Brito DCC, Domingues SFS, Rodrigues APR, Silva LM, Alves KA, Wu X, et al. Betaine-loaded CaCO3 microparticles improve survival of vitrified feline preantral follicles through higher mitochondrial activity and decreased reactive oxygen species. Reprod Fertil Dev. 2020; 32(5): 531-537.