The role of cannabinoid agonists and antagonists on folliculogenesis and evolutionary events in the mouse ovary

Articles in Press

Document Type : Original Article

Authors

1 Department of Anatomical Science, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran

2 Department of Anatomy and Cell Biology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

3 Department of Pathology, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran

4 Kerman Neuroscience Research Center, Neuropharmacology Institute, Kerman University of Medical Sciences, Kerman, Iran

5 Kerman Neuroscience Research Center (KNRC), Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran

10.22038/ijbms.2025.85417.18468

Abstract

Objective(s): Cannabinoids, derivatives of Cannabis sativa L., can activate the endocannabinoid system via two endogenous receptors, CB1 and CB2. This system is crucial in regulating folliculogenesis, fertility, and reproductive function. This study investigated the potential effects of cannabinoid agonists and antagonists on ovarian health and function in female mice.
Materials and Methods: 80 NMRI mice were divided into 10 groups. Treatment groups received CB1 or CB2 agonists, antagonists, or their combinations for five days. The animals were then sacrificed, the ovaries were excised and weighed, and their volume was measured. Total RNA was extracted from the left ovary for qPCR analysis, while the right ovary was fixed in Bouin’s solution for histological evaluation following H&E staining.
Results: Treatment with CB1/CB2 agonist+CB1 antagonist (W102+AM251) decreased the level of NAPE-PLD (a key factor in the production of endocannabinoids in cells) and increased the level of FAAH (responsible for cannabinoid degradation) genes compared to all groups. CB2 antagonist (AM630) increased the number of primary, preantral, and antral follicles, the volume and weight of ovaries, and estrogen levels. Meanwhile, the CB1 antagonist (AM251) significantly increased microvascular density in the ovaries.
Conclusion: Cannabinoids modulate ovarian physiology and folliculogenesis, with CB2 receptors playing a particularly significant role. Antagonism at CB2 appeared to differentially affect cannabinoid-metabolizing enzymes in ovarian follicles and differentially affect their maturation. However, our preliminary novel findings in mice require human studies before clinical application. 

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References

1. Walker OLS, Holloway AC, Raha S. The role of the endocannabinoid system in female reproductive tissues. J Ovarian Res 2019; 12:1-10.
2. Sampson PB. Phytocannabinoid pharmacology: Medicinal properties of Cannabis sativa constituents aside from the “Big Two”. J Nat Prod 2020; 84:142-160.
3. ElSohly MA, Radwan MM, Gul W, Chandra S, Galal A. Phytochemistry of Cannabis sativa L. Phytocannabinoids 2017:1-36.
4. Zhu N, Janssen AP, van der Stelt M. Understanding and targeting the endocannabinoid system with activity‐based protein profiling. Isr J Chem 2023; 63:e202200115.
5. Oyagawa CR, Grimsey NL. Cannabinoid receptor CB1 and CB2 interacting proteins: Techniques, progress and perspectives. Methods Cell Biol 2021; 166:83-132.
6. Han L, Xu S, Zhou D, Chen R, Ding Y, Zhang M, et al. Unveiling the causal link between metabolic factors and ovarian cancer risk using Mendelian randomization analysis. Front Endocrinol 2024; 15:1401648.
7. Chanda D, Neumann D, Glatz JF. The endocannabinoid system: Overview of an emerging multi-faceted therapeutic target. Prostaglandins Leukot Essent Fatty Acids 2019; 140:51-56.
8. Sun Y, Zong C, Liu J, Zeng L, Li Q, Liu Z, et al. C-myc promotes miR-92a-2-5p transcription in rat ovarian granulosa cells after cadmium exposure. Toxicol Appl Pharmacol 2021; 421:115536.
9. El-Talatini MR, Taylor AH, Elson JC, Brown L, Davidson AC, Konje JC. Localisation and function of the endocannabinoid system in the human ovary. PLoS One 2009; 4:e4579.
10. Gammon CM, Freeman GM, Jr., Xie W, Petersen SL, Wetsel WC. Regulation of gonadotropin-releasing hormone secretion by cannabinoids. Endocrinology 2005; 146:4491-4499.
11. Walker OLS, Holloway AC, Raha S. The role of the endocannabinoid system in female reproductive tissues. J Ovarian Res 2019; 12:3-12.
12. Albano F. 73rd Congress of the Italian Society of Pediatrics. Ital J Pediatr 2017; 43:110-151.
13. Maia J, Fonseca B, Teixeira N, Correia-da-Silva G. The fundamental role of the endocannabinoid system in endometrium and placenta: Implications in pathophysiological aspects of uterine and pregnancy disorders. Hum Reprod Update 2020; 26:586-602.
14. Cecconi S, Rapino C, Di Nisio V, Rossi G, Maccarrone M. The (endo) cannabinoid signaling in female reproduction: What are the latest advances? Prog Lipid Res 2020; 77:101019.
15. Fonseca BM, Rebelo I. Cannabis and cannabinoids in reproduction and fertility: Where we stand. Reprod Sci 2022; 29:2429-2439.
16. Karasu T, Marczylo T, Maccarrone M, Konje J. The role of sex steroid hormones, cytokines and the endocannabinoid system in female fertility. Hum Reprod Update 2011; 17:347-361.
17. Maccarrone M, Rapino C, Francavilla F, Barbonetti A. Cannabinoid signalling and effects of cannabis on the male reproductive system. Nat Rev Urol 2021; 18:19-32.
18. Scotchie JG, Savaris RF, Martin CE, Young SL. Endocannabinoid regulation in human endometrium across the menstrual cycle. Reprod Sci 2015; 22:113-123.
19. Struik D, Sanna F, Fattore L. The modulating role of sex and anabolic-androgenic steroid hormones in cannabinoid sensitivity. Front Behav Neurosci 2018:249-262.
20. Soltysik K, Czekaj P. Membrane estrogen receptors-is it an alternative way of estrogen action. J Physiol Pharmacol 2013; 64:129-142.
21. Rubino T, Parolaro D. Sexually dimorphic effects of cannabinoid compounds on emotion and cognition. Front Behav Neurosci 2011; 5:64-68.
22. Maia J, Almada M, Silva A, Correia-da-Silva G, Teixeira N, Sá S, et al. The endocannabinoid system expression in the female reproductive tract is modulated by estrogen. J Steroid Biochem Mol Biol 2017; 174:40-47.
23. Ayakannu T, Taylor AH, Konje JC. Cannabinoid receptor expression in estrogen-dependent and estrogen-independent endometrial cancer. J Recept Signal Transduct Res 2018; 38:385-392.
24. Wu N, Zhang X, Fang C, Zhu M, Wang Z, Jian L, et al. Progesterone enhances niraparib efficacy in ovarian cancer by promoting palmitoleic-acid-mediated ferroptosis. Res (Wash D C) 2024; 7:0371.
25. Kumar MS, Chen CL. Effect of an acute dose of delta 9-THC on hypothalamic luteinizing hormone releasing hormone and met-enkephalin content and serum levels of testosterone and corticosterone in rats. Subst Alcohol Actions Misuse 1983; 4:37-43.
26. Tyrey L. δ-9-Tetrahydrocannabinol suppression of episodic luteinizing hormone secretion in the ovariectomized rat. Endocrinology 1978; 102:1808-1814.
27. Dobovišek L, Hojnik M, Ferk P. Overlapping molecular pathways between cannabinoid receptors type 1 and 2 and estrogens/androgens on the periphery and their involvement in the pathogenesis of common diseases (Review). Int J Mol Med 2016; 38:1642-1651.
28. Sun X, Dey SK. Synthetic cannabinoids and potential reproductive consequences. Life Sci 2014; 97:72-77.
29. Carvalho RK, Souza MR, Santos ML, Guimarães FS, Pobbe RLH, Andersen ML, et al. Chronic cannabidiol exposure promotes functional impairment in sexual behavior and fertility of male mice. Reprod Toxicol 2018; 81:34-40.
30. Craft RM, Marusich JA, Wiley JL. Sex differences in cannabinoid pharmacology: A reflection of differences in the endocannabinoid system? Life Sci 2013; 92:476-481.
31. Dow-Edwards D. Sex differences in the interactive effects of early life stress and the endocannabinoid system. Neurotoxicol Teratol 2020; 80:106893.
32. Cora MC, Kooistra L, Travlos G. Vaginal cytology of the laboratory rat and mouse: review and criteria for the staging of the estrous cycle using stained vaginal smears. Toxicol Pathol 2015; 43:776-793.
33. Marcondes F, Bianchi F, Tanno A. Determination of the estrous cycle phases of rats: some helpful considerations. Braz J Biol 2002; 62:609-614.
34. Greish K, Mathur A, Al Zahrani R, Elkaissi S, Al Jishi M, Nazzal O, et al. Synthetic cannabinoids nano-micelles for the management of triple negative breast cancer. J Control Release 2018; 291:184-195.
35. Gatley SJ, Gifford AN, Volkow ND, Lan R, Makriyannis A. 123I-labeled AM251: A radioiodinated ligand which binds in vivo to mouse brain cannabinoid CB1 receptors. Eur J Pharmacol 1996; 307:331-338.
36. Sufian MS, Amin MR, Ali DW. Early suppression of the endocannabinoid degrading enzymes FAAH and MAGL alters locomotor development in zebrafish. J Exp Biol 2021; 224:jeb242635.
37. Pertwee R, Griffin G, Fernando S, Li X, Hill A, Makriyannis A. AM630, a competitive cannabinoid receptor antagonist. Life Sci 1995; 56:1949-1955.
38. Asiri YI, Fung T, Schwarz SK, Asseri KA, Welch ID, Schuppli CA, et al. An intraplantar hypertonic saline assay in mice for rapid screening of analgesics. Anesth Analg 2018; 127:548-555.
39. Li A-L, Carey LM, Mackie K, Hohmann AG. Cannabinoid CB2 agonist GW405833 suppresses inflammatory and neuropathic pain through a CB1 mechanism that is independent of CB2 receptors in mice. J Pharmacol Exp Ther 2017; 362:296-305.
40. Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative CT method. Nat Protoc 2008; 3:1101-1108.
41. Nematollahi-Mahani SN, Ganjalikhan-Hakemi S, Abdi Z. The regression of glioblastoma multiforme is time dependent in the wild-type rat xenograft model. Basic Clin Neurosci 2023; 14:263-272.
42. Musanejad E, Haghpanah T, Mirzaie V, Ezzatabadipour M. Effects of ethanol and nicotine co-administration on follicular atresia and placental histo-morphology in the first-generation mice pups during intrauterine development and lactation periods. Toxicol Rep 2021; 8:793-803.
43. Wood GA, Fata JE, Watson KL, Khokha R. Circulating hormones and estrous stage predict cellular and stromal remodeling in murine uterus. Reproduction 2007; 133:1035-1044.
44. Reggio PH. Endocannabinoid binding to the cannabinoid receptors: What is known and what remains unknown. Curr Med Chem 2010; 17:1468-1486.
45. Totorikaguena L, Olabarrieta E, Lolicato F, Romero‐Aguirregomezcorta J, Smitz J, Agirregoitia N, et al. The endocannabinoid system modulates the ovarian physiology and its activation can improve in vitro oocyte maturation. J Cell Physiol 2020; 235:7580-7591.
46. Atakan Z. Cannabis, a complex plant: different compounds and different effects on individuals. Ther Adv Psychopharmacol 2012; 2:241-254.
47. Green B, Kavanagh D, Young R. Being stoned: A review of self‐reported cannabis effects. Drug Alcohol Rev 2003; 22:453-460.
48. Cohen K, Weinstein A. The effects of cannabinoids on executive functions: Evidence from cannabis and synthetic cannabinoids—a systematic review. Brain Sci 2018; 8:40-58.
49. Cohen K, Weizman A, Weinstein A. Positive and negative effects of cannabis and cannabinoids on health. Clin Pharmacol Ther 2019; 105:1139-1147.
50. Sun D, Li X, Nie S, Liu J, Wang S. Disorders of cancer metabolism: The therapeutic potential of cannabinoids. Biomed Pharmacother 2023; 157:113993.
51. Bagavandoss P, Grimshaw S. Temporal and spatial distribution of the cannabinoid receptors (CB1, CB2) and Fatty acid amide hydroxylase in the rat ovary. Reprod Biol 2010; 293:1425-1432.
52. Lim J, Lee H-L, Nguyen J, Shin J, Getze S, Quach C, et al. Adolescent exposure to low-dose Δ9-tetrahydrocannabinol depletes the ovarian reserve in female mice. Toxicol Sci 2023; 193:31-47.
53. Li X, Wang Y-K, Song Z-Q, Du Z-Q, Yang C-X. Dimethyl sulfoxide perturbs cell cycle progression and spindle organization in porcine meiotic oocytes. PLoS One 2016; 11:e0158074.
54. Cecconi S, Capacchietti G, Russo V, Berardinelli P, Mattioli M, Barboni B. In vitro growth of preantral follicles isolated from cryopreserved ovine ovarian tissue. Biol Reprod 2004; 70:12-17.
55. Pirone A, Lenzi C, Briganti A, Abbate F, Levanti M, Abramo F, et al. Spatial distribution of cannabinoid receptor 1 and fatty acid amide hydrolase in the cat ovary and oviduct. Acta Histochem 2017; 119:417-422.
56. El-Talatini MR, Taylor AH, Elson JC, Brown L, Davidson AC, Konje JCJPO. Localisation and function of the endocannabinoid system in the human ovary. PLoS One 2009; 4:e4579.
57. Tahermanesh K, Hakimpour S, Govahi A, Keyhanfar F, Kashi AM, Chaichian S, et al. Treatment of ovarian hyperstimulation syndrome in a mouse model by cannabidiol, an angiogenesis pathway inhibitor. Biomed Res Int 2022; 2022:1111777.
Iranian Journal of Basic Medical Sciences

Articles in Press, Accepted Manuscript
Available Online from 18 May 2025

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