Synergic effect of bee pollen and metformin on proliferation and apoptosis of granulosa cells: Rat model of polycystic ovary syndrome
Leila Naseri
Department of Anatomical Sciences, Student Research Committee, Kermanshah University of Medical Sciences, Kermanshah, Iran
Contribution: Writing - original draft
Search for more papers by this authorMohammad Rasool Khazaei
Department of Molecular Medicine, Fertility and Infertility Research Center, Research Institute for Health Technology, Kermanshah University of Medical Sciences, Kermanshah, Iran
Contribution: Writing - review & editing
Search for more papers by this authorCorresponding Author
Mozafar Khazaei
Department of Anatomical Sciences, Fertility and Infertility Research Center, Research Institute for Health Technology, Kermanshah, Iran
Correspondence
Mozafar Khazaei, Department of Anatomical Sciences, Fertility and Infertility Research Center, Research Institute for Health Technology, University Ave, Shahid Shirodi Blvd., P.O. Box: 6714869914, Kermanshah, Iran.
Email: [email protected]
Contribution: Writing - review & editing
Search for more papers by this authorLeila Naseri
Department of Anatomical Sciences, Student Research Committee, Kermanshah University of Medical Sciences, Kermanshah, Iran
Contribution: Writing - original draft
Search for more papers by this authorMohammad Rasool Khazaei
Department of Molecular Medicine, Fertility and Infertility Research Center, Research Institute for Health Technology, Kermanshah University of Medical Sciences, Kermanshah, Iran
Contribution: Writing - review & editing
Search for more papers by this authorCorresponding Author
Mozafar Khazaei
Department of Anatomical Sciences, Fertility and Infertility Research Center, Research Institute for Health Technology, Kermanshah, Iran
Correspondence
Mozafar Khazaei, Department of Anatomical Sciences, Fertility and Infertility Research Center, Research Institute for Health Technology, University Ave, Shahid Shirodi Blvd., P.O. Box: 6714869914, Kermanshah, Iran.
Email: [email protected]
Contribution: Writing - review & editing
Search for more papers by this authorAbstract
Polycystic ovary syndrome (PCOS) is the most common cause of infertility associated with metabolic complications. Several classes of pharmacological agents have been used to manage PCOS. These drugs have shown adverse effects. Various studies showed the bee pollen (BP) as a substance rich in phytoestrogens. This study aimed to investigate the effects of BP and metformin alone and in combination with proliferation and apoptosis of granulosa cells in the rat model of PCOS. In this experimental study, 54 Wistar rats (180–210 g), was injected 2 mg of estradiol valerate intramuscularly and six rats were considered as control. After 60 days, the rats were divided into control, sham, and experimental groups. The rats were treated with bee pollen (50, 100, and 200 mg/kg) and metformin (300 mg/kg), either individually or in combination. Ovarian histology assessment was examined by H&E staining. The serum levels of NO and TNF-α were evaluated. The expressions of P53 and Ki67 were measured by IHC. In the BP and metformin-treated PCOS group, the preantral and antral follicles increased, and cystic follicles significantly decreased (p < .01). The levels of TNF-α, NO, as well as the expressions of Ki67 were decreased in the treated groups compared to the PCOS group (p < .01). On the contrary, apoptosis increased in the groups treated with BP compared to the untreated group (p < .01). BP individually or synergistically with metformin improved the symptoms of PCOS.
CONFLICT OF INTEREST
The authors declare no conflict of interest in this study and they are responsible for the content and writing of the paper.
REFERENCES
- Abdel-Gawad, E. I. (2010). Potential impact of bee pollen administration during pregnancy in rats. Journal of American Science, 6(5), ISSN: 1545–1003.
- Abtahi-Eivari, S. H., Moghimian, M., Soltani, M., Shoorei, H., Asghari, R., Hajizadeh, H., Shokoohi, M., Alami, S., & Khalife Ghaderi, F. (2018). The effect of galega officinalis on hormonal and metabolic profile in a rat model of polycystic ovary syndrome. International Journal of Womens Health and Reproduction Sciences, 6(3), 276–282. https://doi.org/10.15296/ijwhr.2018.46
- Bottini, A., Berruti, A., Bersiga, A., Brizzi, M. P., Bruzzi, P., Aguggini, S., Brunelli, A., Bolsi, G., Allevi, G., Generali, D., Betri, E., Bertoli, G., Alquati, P., & Dogliotti, L. (2001). Relationship between tumour shrinkage and reduction in ki67 expression after primary chemotherapy in human breast cancer. British Journal of Cancer, 85(8), 1106–1112. https://doi.org/10.1054/Bjoc.2001.2048
- Braillon, A. (2005). Polycystic ovary syndrome. New England Journal of Medicine, 352(26), 2756. https://doi.org/10.1056/Nejmra041536
- Chen, J., Shen, S., Tan, Y., Xia, D., Xia, Y., Cao, Y., Wang, W., Wu, X., Wang, H., Yi, L., Gao, Q., & Wang, Y. (2015). The correlation of aromatase activity and obesity in women with or without polycystic ovary syndrome. Journal of Ovarian Research, 8(1), 11. https://doi.org/10.1186/s13048-015-0139-1
- Corakci, A., Filiz, S., Caliskan, E., Dalcik, C., Ozeren, S., & Dalcik, H. (2005). The effects of ovulation induction on ovarian epithelium dysplasia scores and Ki67 expression: An experimental study on rats. International Journal of Gynecologic Cancer, 15(5), 866–871. https://doi.org/10.1111/J.1525-1438
- Das, M., Djahanbakhch, O., Hacihanefioglu, B., Saridogan, E., Ikram, M., Ghali, L., Raveendran, M., & Storey, A. (2008). Granulosa cell survival and proliferation are altered in polycystic ovary syndrome. Journal of Clinical Endocrinology and Metabolism, 93(3), 881–887. https://doi.org/10.1210/Jc.2007-1650
- De Leo, V., La Marca, A., Ditto, A., Morgante, G., & Cianci, A. (1999). Effects of metformin on gonadotropin-induced ovulation in women with polycystic ovary syndrome. Fertility and Sterility, 72(2), 282–285. https://doi.org/10.1016/s0015-0282(99)00208-3
- Denisow, B., & Denisow-Pietrzyk, M. (2016). Biological and therapeutic properties of bee pollen: A review. Journal of the Science of Food and Agriculture, 96(13), 4303–4309. https://doi.org/10.1002/Jsfa.7729
- Fleming, R., Hopkinson, Z. E., Wallace, A. M., Greer, I. A., & Sattar, N. (2002). Ovarian function and metabolic factors in women with oligomenorrhea treated with metformin in a randomized double blind placebo-controlled trial. The Journal of Clinical Endocrinology & Metabolism, 87(2), 569–574. https://doi.org/10.1210/Jcem.87.2.8261
- Franks, S., & Hardy, K. (2020). What causes anovulation in PCOS? Current Opinion in Endocrine and Metabolic Research, 297–302. https://doi.org/10.1111/J.1365-2605.2005.00623
- Ghanbari, E., Khazaei, M. R., Khazaei, M., & Nejati, V. (2018). Royal jelly promotes ovarian follicles growth and increases steroid hormones in immature rats. International Journal of Fertility & Sterility, 11(4), 263–269. https://doi.org/10.22074/Ijfs.2018.5156
- Heibashy, M., Mazen, G., & Shahin, M. (2013). Metabolic changes and hormonal disturbances in polycystic ovarian syndrome rats and the amelioration effects of metformin and/or cinnamon extraction. Journal of American Science, 9(12), 54–P62. https://doi.org/10.1016/J.Molmet.2020.01.001
- Ibáñez, L., Valls, C., & De Zegher, F. (2006). Discontinuous low-dose flutamide–metformin plus an oral or a transdermal contraceptive in patients with hyperinsulinaemic hyperandrogenism: Normalizing effects on CRP, TNF-α and the neutrophil/lymphocyte ratio. Human Reproduction, 21(2), 451–456. https://doi.org/10.1093/Humrep/Dei329
- Jablonka-Shariff, A., & Olson, L. M. (1998). The role of nitric oxide in oocyte meiotic maturation and ovulation: Meiotic abnormalities of endothelial nitric oxide synthase knock-out mouse oocytes. Endocrinology, 139(6), 2944–2954. https://doi.org/10.1210/Endo.139.6.6054
- Jakubowska, J., Bohdanowicz-Pawlak, A., Milewicz, A., Szymczak, J., Bednarek-Tupikowska, G., & Demissie, M. (2008). Plasma cytokines in obese women with polycystic ovary syndrome, before and after metformin treatment. Gynecological Endocrinology, 24(7), 378–384. https://doi.org/10.1080/09513590802128968
- Johansson, J., & Stener-Victorin, E. (2013). Polycystic ovary syndrome: Effect and mechanisms of acupuncture for ovulation induction. Evidence-Based Complementary and Alternative Medicine, 2013, 762615. https://doi.org/10.1155/2013/762615
- Karimi Jashni, H., Kargar Jahromi, H., & Bagheri, Z. (2016). The effect of palm pollen extract on polycystic ovary syndrome (POS) in rats. International Journal of Medical Research & Health Sciences, 5(5), 317–321.ISSN:2319–5886.
- Kimura, Y., Ito, H., Ohnishi, R., & Hatano, T. (2010). Inhibitory effects of polyphenols on human cytochrome P450 3A4 and 2C9 activity. Food and Chemical Toxicology, 48(1), 429–435. https://doi.org/10.1016/j.fct.2009.10.041
- King, L. A., Okagaki, T., Gallup, D. G., Twiggs, L. B., Messing, M. J., & Carson, L. F. (1996). Mitotic count, nuclear atypia, and immunohistochemical determination of Ki-67, C-Myc, P21-Ras, C-Erbb2, and P53 expression in granulosa cell tumors of the ovary: Mitotic count and Ki-67 are indicators of poor prognosis. Gynecologic Oncology, 61(2), 227–232. https://doi.org/10.1006/Gyno.1996.0130
- Kohzadi, M., Khazaei, M. R., Choobsaz, F., & Khazaei, M. (2020). Relationship between serum levels of anti-mullerian hormone, adiponectin and oxidative stress markers in patients with polycystic ovary syndrome. International Journal of Fertility & Sterility, 14(1), 27. https://doi.org/10.22074/Ijfs.2020.5809
- Malini, T., & Vanithakumari, G. (1991). Antifertility effects of β-sitosterol in male albino rats. Journal of Ethnopharmacology, 35(2), 149–153. https://doi.org/10.1016/0378-8741(91)90066-M
- Mokhtari, M., Ebrahimpoor, M. R., & Harfsheno, S. (2014). The effects of alcoholic extract of marrubum vulgare on hormonal parameters in female rat model of polycystic ovarian syndrome. Medical Science Journal of Islamic Azad Univesity-Tehran Medical Branch, 24(2), 74–80.
- Naseri, L., Khazaei, M. R., & Khazaei, M. (2020). Potential therapeutic effect of bee pollen and metformin combination on testosterone and estradiol levels, apoptotic markers and total antioxidant capacity in rats model of polycystic ovary syndrome. International Journal of Fertility and Sterility. https://doi.org/10.22074/IJFS.2020.134604
- Nestler, J. E., Powers, L. P., Matt, D. W., Steingold, K. A., Plymate, S. R., Rittmaster, R. S., Clore, J. N., & Blackard, W. G. (1991). A direct effect of hyperinsulinemia on serum sex hormone-binding globulin levels in obese women with the polycystic ovary syndrome. The Journal of Clinical Endocrinology & Metabolism, 72(1), 83–89. https://doi.org/10.1210/jcem-72-1-83
- Quaas, A. M., & Legro, R. S. (2019). Pharmacology of medications used for ovarian stimulation. Best Practice & Research Clinical Endocrinology & Metabolism, 33(1), 21–33. https://doi.org/10.1016/J.Beem.2018.10.002
- Rencber, S. F., Ozbek, S. K., Eraldemır, C., Sezer, Z., Kum, T., Ceylan, S., & Guzel, E. (2018). Effect of resveratrol and metformin on ovarian reserve and ultrastructure in PCOS: An experimental study. Journal of Ovarian Research, 11(1), 55. https://doi.org/10.1186/S13048-018-0427-7
- Rzepecka-Stojko, A., Stojko, J., Kurek-Górecka, A., Górecki, M., Kabała-Dzik, A., Kubina, R., Moździerz, A., & Buszman, E. (2015). Polyphenols from bee pollen: Structure, absorption, metabolism and biological activity. Molecules, 20(12), 21732–21749. https://doi.org/10.3390/molecules201219800
- Saadati, N., Yaghmaei, P., Hashemi, F., Ramezani Tehrani, F., & Hedayati, M. (2012). Association of serum omentin levels in women with polycystic ovarian syndrome. Iranian Journal of Endocrinology and Metabolism, 14(4), 375–379.
- Sajadimajd, S., & Khazaei, M. (2018). Oxidative stress and cancer: The role of Nrf2. Current Cancer Drug Targets, 18(6), 538–557. https://doi.org/10.2174/15680096176661710021144228
- Samy, N., Hashim, M., Sayed, M., & Said, M. (2009). Clinical significance of inflammatory markers in polycystic ovary syndrome: Their relationship to insulin resistance and body mass index. Disease Markers, 26(4), 163–170. https://doi.org/10.3233/DMA-2009-0627
- Silverman, M. N., Pearce, B. D., Biron, C. A., & Miller, A. H. (2005). Immune modulation of the hypothalamic-pituitary-adrenal (HPA) axis during viral infection. Viral Immunology, 18(1), 41–78. https://doi.org/10.1089/Vim.2005.18.41
- Stubbs, S. A., Stark, J., Dilworth, S. M., Franks, S., & Hardy, K. (2007). Abnormal preantral folliculogenesis in polycystic ovaries is associated with increased granulosa cell division. Journal of Clinical Endocrinology and Metabolism, 92(11), 4418–4426. https://doi.org/10.1210/Jc.2007-0729
- Tao, X., Cai, L., Chen, L., Ge, S., & Deng, X. (2019). Effects of metformin and exenatide on insulin resistance and Ampkα-SIRT1 molecular pathway in PCOS rats. Journal of Ovarian Research, 12(1), 1–8. https://doi.org/10.1186/s13048-019-055508
- Thakur, M., & Nanda, V. (2020). Composition and functionality of bee pollen: A review. Trends in Food Science & Technology, 98, 82–106. https://doi.org/10.1016/J.Tifs.2020.02.001
- Vendola, K. A., Zhou, J., Adesanya, O. O., Weil, S. J., & Bondy, C. A. (1998). Androgens stimulate early stages of follicular growth in the primate ovary. The Journal of Clinical Investigation, 101(12), 2622–2629. https://doi.org/10.1172/JCI2081
- Whittles, C. E., Biddlestone, L. R., Burton, A., Barr, H., Jankowski, J. A., Warner, P. J., & Shepherd, N. A. (1999). Apoptotic and proliferative activity in the neoplastic progression of barrett's oesophagus: A comparative study. The Journal of Pathology, 187(5), 535–540. https://doi.org/10.1002/(Sici)1096-9896(199904)187:5<535::Aid-Path302>3.0.Co;2-G
10.1002/(SICI)1096-9896(199904)187:5<535::AID-PATH302>3.0.CO;2-G CAS PubMed Web of Science® Google Scholar
