Ethanol Extract from Yellow Pumpkin Modulate Hormonal Levels and Ovarian Follicle Dynamics in Hypoestrogenic Wistar Rats
Article Sidebar
Main Article Content
Abstract
Depo Medroxyprogesterone Acetate (DMPA) is a progesterone analog that induces a hypoestrogenic response by inhibiting gonadotropin-releasing hormone (GnRH) needed for the release of follicle-stimulating hormone (FSH) and Luteinizing Hormone (LH), thereby preventing ovulation, decreasing the number of antral follicles, and suppressing stromal and epithelial cell proliferation. Side effects of DMPA can be reduced by using natural ingredients such as yellow pumpkin seeds (Cucurbita pepo L.). This study aimed to determine the effect of pumpkin seed ethanol extract on FSH levels and activation of Ki-67 expression in endometrial stromal cells, beta estrogen receptors, antral follicles, stromal cells, and endometrial epithelial cells in hypoestrogen type Wistar rats with DMPA injection. This in vivo study used 25 female rats (Rattus norvegicus) that were randomly divided into five groups: a control group and four treatment groups. The control group, without DMPA (sterile aquadest injection), as well as the treatment groups, were injected with a dose of 2.7 mg/kg bw of DMPA every 3 days for 21 days. Ovarian FSH levels were evaluated by ELISA, Ki-67, and ERβ expression by immunohistochemistry (IHC), antral follicle count, stromal cells, and endometrial epithelium by Hematoxylin Eosin (HE). The findings of the study revealed significant differences and correlations between extract dose and FSH levels, Ki-67 expression, ER-beta, antral follicle count, stromal cells, and endometrial epithelial cells. The result of the study showed that yellow pumpkin seed ethanol extract in various doses enhanced FSH levels, Ki-67 expression, ER-beta, antral follicle count, stromal cell count, and endometrial epithelial cell count compared to a positive control group exposed only to DMPA.
Downloads
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
References
1. Kemenkes RI. Indonesia Health Profile. (Indonesia KKR, ed.).; 2018. Primadi O, Budijanto D, Kurniawan R, Yudianto, Hardhana B, Siswanti T. Indonesia health profile. (edisi 1). Indonesia: Ministry of Health, Republic of Indonesia; 2018. 394 p
2. Sukatin, Nurkhalipah, Kurnia A, Ramadani D, Fatimah. The relationship between the use of 3-month DMPA birth control injections and side effects in birth control acceptors at the Teluk Bayur Health Center. Ind Mul Sci J. IJOMS. 2022;1(9):1278-1285.
3. Veri N, Mutiah C, Dewita D, Heniwati H, Fazdria F, Lajuna L, Salfiyadi T S. The effect of duration of use of depomedroxyprogesterone acetate on the thickness of the vaginal epithelium of mice. Med Sci J. 2021;9(A):73-77. doi:10.3889/oamjms.2021.5574
4. Nurhayati. Medroxy Progesterone Acetate (DMPA) Depot and Menstrual Cycle Disorders. In: Dewi YF, ed. Medroxy Progesterone Acetate (DMPA) Depot Monograph and Menstrual Cycle Disorders. 1st ed. PT. Pena Persada Kerta Utama; 2022:11-61.
5. Liu Y, Li X, Shen X, Chen Q, Yu S, Wang Y, Long H, Wang L, Liu Q, Kuang Y, and O’Byrne K T. Dynorphin and GABAA receptor signaling contribute to progesterone’s inhibition of the LH surge in female mice. Endocrinology. 2020;161(5):1-10. doi:10.1210/endocr/bqaa036
6. Paszkowski T, Bińkowska M, Dȩbski R, Krzyczkowska-Sendrakowska M, Skrzypulec-Plinta V, Zgliczyński W. Menopausal hormone therapy in questions and answers - A manual for physicians of various specialties. Przeglad Menopauzalny. 2019;18(1):1-8. doi:10.5114/pm.2019.84150
7. Lestari B, Meiyanto E. A Review: The Emerging Nutraceutical Potential of Pumpkin Seeds. Indones. J. Cancer Chemoprevention. 2018;9(2):92. doi:10.14499/indonesianjcanchemoprev9iss2pp92-101
8. Avenant C, Bick AJ, Skosana SB, Mandisa SM, Justus HG, Jenni S, Mags B, Ivana B, Ishen S, Joanne B, Pai Lien C, Karl Heinz S, Donita A, and Janet P. H. Misreporting contraceptive use and the association of peak study progestin levels with weight and BMI among women randomized to the progestin only injectable contraceptives DMPA-IM and NET-EN. PLoS One. 2023;18 doi:10.1371/journal.pone.0295959
9. Zhang Q, He S, Meng Y, and Saijiao L. Effect of medroxyprogesterone acetate dose in progestin-primed ovarian stimulation on pregnancy outcomes in poor ovarian response patients with different body mass index levels. Front Endocrinol (Lausanne). 2024;15:1-8.
doi:10.3389/fendo.2024.1352522
10. Oh J, Hong S, Ko SH, Kim HS. Evaluation of Antioxidant Effects of Pumpkin (Cucurbita pepo L.) Seed Extract on Aging and Menopause Related Diseases Using Saos-2 Cells and Ovariectomized Rats. Antioxidants. 2024;13(2). doi:10.3390/antiox13020241
11. Domínguez-López I, Yago-Aragón M, Salas-Huetos A, Tresserra-Rimbau A, Hurtado-Barroso S. Effects of dietary phytoestrogens on hormones throughout a human lifespan: A review. Nutrients. 2020;12(8):1-25. doi:10.3390/nu12082456
12. Sathe A, Gerriets V. Medroxyprogesterone. In: Gerriets AS; V, ed. National Center for Biotechnology Information. NCBI; 2023:1-8.
13. Wahyuni ES. Contraceptives. First. (Firrahmawati L, Amin MA, eds.). Pustaka Hanif; 2017.
14. Yanti, Lilis C., and Annisa Lamaindi. "Effect of Use of DMPA Injection Contraception Towards Changes of Menstrual Cycle Disorders in Kb Acceptors." J. Ilm. Kesehat. Sandi Husada. 2021; 10(1):314-318, doi:10.35816/jiskh.v10i1.596.
15. Sun X, Kaufman P. HHS Public Access Ki-67 : more than a proliferation marker. Chromosoma. 2018;127(2):175-186. doi:10.1007/s00412-018-0659-8.Ki-67
16. Delbandi AA, Mahmoudi M, Shervin A, Heidari S, Kolahdouz-Mohammadi R, Zarnani AH. Evaluation of apoptosis and angiogenesis in ectopic and eutopic stromal cells of patients with endometriosis compared to non-endometriotic controls. BMC Womens Health. 2020;20(1):1-9. doi:10.1186/s12905-019-0865-4
17. Meyler’s. Phytoestrogens. Elsevier, Meyler’s Side Effects of Drugs (Sixteenth Edition). 2019;4806:755-757. doi:https://doi.org/10.1016/B978-0-444-53717-1.00151-7
18. Monteseirín J, De La Calle A, Delgado J, Llamas E, and Conde J. Antigen receptor signaling. Allergol Immunopathol (Madr). 2019;24(5):185-192. doi:10.1016/bs.apcsb.2019.01.001.Estrogen
19. Yu K, Huang ZY, Xu XL, Li J, Fu XW, Deng SL. Estrogen Receptor Function: Impact on the Human Endometrium. Front Endocrinol (Lausanne). 2022;13:1-16. doi:10.3389/fendo.2022.827724
20. Firmawati A, Hutabarat MAK, Pratiwi H, Haryo A. Expression of estrogen receptors alpha (ERs α) and folliculogenesis profile in ovary of the rats ovarian hypofunction model. Jurnal Ilmu-Ilmu Peternakan. 2021;31(1):18-26. doi:10.21776/ub.jiip.2021.031.01.03
21. Muflihah IS, Analisawati T, Margiana W, Maulana AM. Differences in Estrogen Levels Before and After Giving Callanggi Tea To Wistar Straining White Female Rats (Rattus Norvegicus) in Medroxyprogesterone Acetate (DMPA) Induction. J. Inter. Multi. Res. Liter. 2023;2(1):105-110. doi:10.53067/ijomral.v2i1.94
22. Avenant C, Bick AJ, Skosana SB, Moliki JM, Madliki MS, Hofmeyr GJ, Smit J, Beksinska M, Beesham I, Seocharan I, Batting J, Chen PL, Storbeck KH, Africander D, Janet, and Hapgood P. Misreporting contraceptive use and the association of peak study progestin levels with weight and BMI among women randomized to the progestin-only injectable contraceptives DMPA-IM and NET-EN. PLoS One. 2023;18. doi:10.1371/journal.pone.0295959
23. Gorzkiewicz J, Bartosz G, Sadowska-Bartosz I. The potential effects of phytoestrogens: The Role in neuroprotection. Molecules. 2021;26(10):1-12. doi:10.3390/molecules26102954
24. Petrine JCP, Del Bianco-Borges B. The influence of phytoestrogens on different physiological and pathological processes: An overview. Phytotherapy Research. 2021;35(1):180-197. doi:10.1002/ptr.6816
25. Tsikouras P, Chalkidou A, Iatrakis G, Aise Chatzi Ismail M, Alexios A, Konstantinos N, Nektaria K, Theopi N, Sonia K, Stefanos Z, and Nikolaos N. The Contribution of Isoflavones in Menopausal Symptomatic as Alternative Treatment Option. Women’s Health Problems - A Global Perspective. IntechOpen; 2024. Available from: http://dx.doi.org/10.5772/intechopen.114212
26. Kawakita T, Yasui T, Yoshida K, Matsui S, Iwasa T. Associations of LH and FSH with reproductive hormones depending on each stage of the menopausal transition. BMC Womens Health. 2023;23(1):1-9. doi:10.1186/s12905-023-02438-5
27. Zhang J, Xu Y, Liu H, Pan Z. MicroRNAs in ovarian follicular atresia and granulosa cell apoptosis. Reprod Biol Endocrinol. 2019;17(1):1-11. doi:10.1186/s12958-018-0450-y
28. Echeverria V, Echeverria F, Barreto GE, Echeverría J, Mendoza C. Estrogenic Plants: to Prevent Neurodegeneration and Memory Loss and Other Symptoms in Women After Menopause. Front Pharmacol. 2021;12(May):1-25. doi:10.3389/fphar.2021.644103
29. Zalenskaya IA, Chandra N, Yousefieh N, Sharon MA, Christine KM, Jill LS, Andrea RT, and Gustavo FD. Use of contraceptive depot medroxyprogesterone acetate is associated with impaired cervicovaginal mucosal integrity. J. Clin. Investig. 2018;128(10):4622-4638. doi:10.1172/JCI120583
30. Queckbörner S, Syk Lundberg E, Gemzell-Danielsson K, Davies LC. Endometrial stromal cells exhibit a distinct phenotypic and immunomodulatory profile. Stem Cell Res Ther. 2020;11(1):1-15. doi:10.1186/s13287-019-1496-2
31. Delbandi AA, Mahmoudi M, Shervin A, Heidari S, Kolahdouz-Mohammadi R, Zarnani AH. Evaluation of apoptosis and angiogenesis in ectopic and eutopic stromal cells of patients with endometriosis compared to non-endometriotic controls. BMC Womens Health. 2020;20(1):1-9. doi:10.1186/s12905-019-0865-4
32. Yu K, Huang ZY, Xu XL, Li J, Fu XW, Deng SL. Estrogen Receptor Function: Impact on the Human Endometrium. Front Endocrinol (Lausanne). 2022;13:1-16. doi:10.3389/fendo.2022.827724
33. Dotto JM, Chacha JS. The potential of pumpkin seeds as a functional food ingredient: A review: Biofunctional ingredients of pumpkin seeds. Sci Afr. 2020;10:e00575. doi:10.1016/j.sciaf.2020.e00575
34. Mostrom M, Evans TJ. Phytoestrogens. Elsevier Inc.; 2011. doi:10.1016/B978-0-12-382032-7.10052-9
35. Primiani CN, Pujiati. Potential Estrogenic Pigeon Pea (Cajanus Cajan) On Uterus And Bone Tissue Structure Of Rat Female. National Seminar on Biodiversity VI. 2016;(September 2016).
36. Veri N, Mutiah C, Dewita D, and Salfiyadi T.S. The effect of duration of use of depomedroxyprogesterone acetate on the thickness of the vaginal epithelium of mice. Open Access Maced J Med Sci. 2021;9(A):73-77. doi:10.3889/oamjms.2021.5574
37. Fedotcheva TA, Fedotcheva NI, Shimanovsky NL. Progestins as anticancer drugs and chemosensitizers, new targets and applications. Pharmaceutics. 2021;13(10):1-21. doi:10.3390/pharmaceutics13101616
38. Bradley F, Boger MF, Kaldhusdal V, Lajoie J, Bergstrom S, Omollo K, Damdimopoulos A, Czarnewski P, Månberg A, Oyugi J, Kimani J, Nilsson P, Fowke K, Tjernlund A, and Broliden K. Multi-omics analysis of the cervical epithelial integrity of women using depot medroxyprogesterone acetate. PLoS Pathog. 2022;18(5):1-29. doi:10.1371/journal.ppat.1010494
39. Lindermann Peressoni Teixeira MJ, Colonetti Colombo C, Colonetti L, Rosa MI da, Colonetti T. Influence of phytoestrogens on endometrial thickness: a systematic review and meta-analysis. Climacteric. 2022;25(2):118-127. doi:10.1080/13697137.2021.1921728
40. Hadiningsih EF, Ardela MP, Suryanti S, Nurseta T, Noorhamdani, Winarsih S, and Anita KW, Angelina A. The effect of Bengkuang (Pachyrhizus erosus) ethanol extract on the number of ovarian follicles, amount of epithelium and endometrium stroma cells in DMPA-treated Rattus norvegicus. AIP Conf Proc. 2020;2231. doi:10.1063/5.0002908