Beneficial effects of Sambucus nigra L. (black elder) as a traditional medicine have been associated with the phytoconstituents including polyphenols, terpenes and lectins. Various antioxidant rich natural products have also been implicated with improvement of reproductive health and fertility, however, the effect of Sambucus nigra on the ovarian cell functions has not been investigated yet. The objectives of the present study were to screen the polyphenols in the elderflower and elderberry extracts, and to examine the secretion activity of steroid hormones 17beta-estradiol and progesterone by human ovarian granulosa cells HGL5 after supplementation of the extracts at a concentration range of 12.5 to 100 microg.ml-1. Qualitative as well as quantitative screening of polyphenols by high-performance liquid chromatography with diode-array detector (HPLC-DAD) analysis revealed rutin to be the most abundant polyphenol in both elderflower and elderberry extracts. In culture, neither elderflower nor elderberry extract caused any significant impact (p>0.05) in cell viability as studied by AlamarBlue assay in comparison to control. However, a dose-dependent stimulation of 17beta-estradiol release was detected by ELISA after supplementation of elderflower (at 50 microg.ml-1; p<0.01) and elderberry (at 100 microg.ml-1; p<0.05) extracts at higher doses used in the study. On the other hand, both elderflower and elderberry extracts stimulated the secretion of progesterone by HGL5 cells at a lower dose (12.5 microg.ml-1; p<0.05), as compared to control. Therefore, elderflower and elderberry extracts may have the potential to regulate steroidogenesis in ovarian cells.

AgroBioTech Research Centre Slovak University of Agriculture in Nitra Nitra Slovak Republic Department of Animal Physiology Faculty of Biotechnology and Food Sciences Slovak University of Agriculture in Nitra Nitra Slovak Republic

Zobrazit více v PubMed

ABDRAMANOV A, MASSANYI P, SARSEMBAYEVA N, USENBAYEV A, ALIMOV J, TVRDÁ E. The in vitro effect of elderberry (Sambucus nigra) extract on the activity and oxidative profile of bovine spermatozoa. J Microbiol Biotechnol Food Sci. 2017;6:1319–1322. doi: 10.15414/jmbfs.2017.6.6.1319-1322. DOI

AI A, TANG Z, LIU Y, YU S, LI B, HUANG H, WANG X, CAO Y, ZHANG W. Characterization and identification of human immortalized granulosa cells derived from ovarian follicular fluid. Exp Ther Med. 2019;18:2167–2177. doi: 10.3892/etm.2019.7802. PubMed DOI PMC

ARNHOLD IJ, LOFRANO-PORTO A, LATRONICO AC. Inactivating mutations of luteinizing hormone beta-subunit or luteinizing hormone receptor cause oligo-amenorrhea and infertility in women. Horm Res. 2009;71:75–82. doi: 10.1159/000183895. PubMed DOI

BALDOVSKÁ S, MARUNIAKOVÁ N, SLÁMA P, PAVLÍK A, KOHÚT L, KOLESÁROVÁ A. Efficacy of phytonutrients from pomegranate peel on human ovarian cells in vitro. J Microbiol Biotechnol Food Sci. 2020;10:511–516. doi: 10.15414/jmbfs.2020.10.3.511-516. DOI

BOURAKI G, METALLINOU C, SIMOPOULOU M, CHARALABOPOULOS K, ASIMAKOPOULOS B. Comparison of nine media in the culture of human ovarian granulosa lutein cells. In Vivo. 2012;26:823–825. PubMed

CASTILLO-MALDONADO I, MORENO-ALTAMIRANO MMB, SERRANO-GALLARDO LB. Anti-dengue serotype-2 activity effect of Sambucus nigra leaves- and flowers-derived compounds. Virol Res Rev. 2017;1:1–5. doi: 10.15761/VRR.1000117. DOI

CEDERROTH CR, ZIMMERMANN C, NEF S. Soy, phytoestrogens and their impact on reproductive health. Mol Cell Endocrinol. 2012;355:192–200. doi: 10.1016/j.mce.2011.05.049. PubMed DOI

CHEN H, MIAO Q, GENG M, LIU J, HU Y, TIAN L, PAN J, YANG Y. Anti-tumor effect of rutin on human neuroblastoma cell lines through inducing G2/M cell cycle arrest and promoting apoptosis. Sci World J. 2013;2013:269165. doi: 10.1155/2013/269165. PubMed DOI PMC

CIOCOIU M, MIRON A, MARES L, TUTUNARU D, POHACI C, GROZA M, BADESCU M. The effects of Sambucus nigra polyphenols on oxidative stress and metabolic disorders in experimental diabetes mellitus. J Physiol Biochem. 2009;65:297–304. doi: 10.1007/BF03180582. PubMed DOI

DEROO BJ, KORACH KS. Estrogen receptors and human disease. J Clin Invest. 2006;116:561–570. doi: 10.1172/JCI27987. PubMed DOI PMC

CHOWDHURY S, RAY U, CHATTERJEE BP, ROY SS. Targeted apoptosis in ovarian cancer cells through mitochondrial dysfunction in response to Sambucus nigra agglutinin. Cell Death Dis. 2017;8:e2762. doi: 10.1038/cddis.2017.77. PubMed DOI PMC

DESMAWATI D, SULASTRI D. Phytoestrogens and their health effect. Open Access Maced J Med Sci. 2019;7:495–499. doi: 10.3889/oamjms.2019.086. PubMed DOI PMC

DODE MAN, GRAVES CN. Role of estradiol-17β on nuclear and cytoplasmic maturation of pig oocytes. Anim Reprod Sci. 2003;78:99–110. doi: 10.1016/S0378-4320(03)00080-0. PubMed DOI

GABRIELE M, PUCCI L, ÁRVAY J, LONGO V. Anti-inflammatory and antioxidant effect of fermented whole wheat on TNFα-stimulated HT-29 and NF-κB signaling pathway activation. J Funct Foods. 2018;45:392–400. doi: 10.1016/j.jff.2018.04.029. DOI

HAGAN CR, FAIVRE EJ, LANGE CA. Scaffolding actions of membrane-associated progesterone receptors. Steroids. 2009;74:568–572. doi: 10.1016/j.steroids.2008.12.004. PubMed DOI PMC

HAVELOCK JC, RAINEY WE, CARR BR. Ovarian granulosa cell lines. Mol Cell Endocrinol. 2004;228:67–78. doi: 10.1016/j.mce.2004.04.018. PubMed DOI

HSIEH CY, SANTELL RC, HASLAM SZ, HELFERICH WG. Estrogenic effects of genistein on the growth of estrogen receptor-positive human breast cancer (MCF-7) cells in vitro and in vivo. Cancer Res. 1998;58:3833–3838. PubMed

JEFFERSON WN, WILLIAMS C. Circulating levels of genistein in the neonate, apart from dose and route, predict future adverse female reproductive outcomes. Reprod Toxicol. 2011;31:272–279. doi: 10.1016/j.reprotox.2010.10.001. PubMed DOI PMC

KADASI A, KOLESAROVA A, MARUNIAKOVA N, GROSSMANN R, STOCHMALOVA A, ALEXA R, SIROTKIN AV. The effect of green tea extract - epigallocatechin gallate (EGCG) on porcine ovarian granulosa cell. J Microbiol Biotechnol Food Sci. 2014;3:107–109.

KALTSA O, LAKKA A, GRIGORAKIS S, KARAGEORGOU I, BATRA G, BOZINOU E, LALAS S, MAKRIS DP. A green extraction process for polyphenols from elderberry (Sambucus nigra) flowers using deep eutectic solvent and ultrasound-assisted pretreatment. Molecules. 2020;25:921. doi: 10.3390/molecules25040921. PubMed DOI PMC

KNUDSEN BF, KAACK KV. A review of human health and disease claims for elderberry (Sambucus nigra) fruit. Acta Hortic. 2015;1061:121–131. doi: 10.17660/ActaHortic.2015.1061.12. DOI

KOLESÁROVÁ A, SIROTKIN AV, MELLEN M, ROYCHOUDHURY S. Possible intracellular regulators of female sexual maturation. Physiol Res. 2015;64:379–386. doi: 10.33549/physiolres.932838. PubMed DOI

KOLESAROVA A, ROYCHOUDHURY S, KLINEROVA B, PACKOVA D, MICHALCOVA K, HALENAR M, KOPCEKOVA J, MNAHONCAKOVA E, GALIK B. Dietary bioflavonoid quercetin modulates porcine ovarian granulosa cell functions in vitro. J Environ Sci Health B. 2019;54:533–537. doi: 10.1080/03601234.2019.1586034. PubMed DOI

KRAWITZ C, MRAHEIL MA, STEIN M, IMIRZALIOGLU C, DOMANN E, PLESCHKA S, HAIN T. Inhibitory activity of a standardized elderberry liquid extract against clinically-relevant human respiratory bacterial pathogens and influenza A and B viruses. BMC Complement Altern Med. 2011;11:16. doi: 10.1186/1472-6882-11-16. PubMed DOI PMC

LECOMTE S, DEMAY F, FERRIERE F, PAKDEL F. Phytochemicals targeting estrogen receptors: Beneficial rather than adverse effects? Int J Mol Sci. 2017;18:1381. doi: 10.3390/ijms18071381. PubMed DOI PMC

MAHAJAN DK. Pig model to study dynamics of steroids during ovarian follicular growth and maturation. In: CONN PM, editor. Sourcebook of Models For Biomedical Research. Humana Press; 2008. pp. 425–436. DOI

MAHBOUBI M. Sambucus nigra (black elder) as alternative treatment for cold and flu. Adv Tradit Med. 2020:1–10. doi: 10.1007/s13596-020-00469-z. DOI

MICHALCOVA K, ROYCHOUDHURY S, HALENAR M, TVRDA E, KOVACIKOVA E, VASICEK J, CHRENEK P, BALDOVSKA S, SANISLO L, KREN V, KOLESAROVA A. In vitro response of human ovarian cancer cells to dietary bioflavonoid isoquercitrin. J Environ Sci Health B. 2019;54:752–757. doi: 10.1080/03601234.2019.1633214. PubMed DOI

MŁYNARCZYK K, WALKOWIAK-TOMCZAK D, ŁYSIAK GP. Bioactive properties of Sambucus nigra L as a functional ingredient for food and pharmaceutical industry. J Funct Foods. 2018;40:377–390. doi: 10.1016/j.jff.2017.11.025. PubMed DOI PMC

MOTA AH, DUARTE N, SERRA AT, FERREIRA A, BRONZE MR, CUSTÓDIO L, GASPAR MM, SIMOES S, RIJO P, ASCENSAO L, FAÍSCA P, VIANA AS, PINTO R, KUMAR P, ALMEIDA AJ, REIS CP. Further evidence of possible therapeutic uses of Sambucus nigra L. extracts by the assessment of the in vitro and in vivo anti-inflammatory properties of its PLGA and PCL-based nanoformulations. Pharmaceutics. 2020;12:1181. doi: 10.3390/pharmaceutics12121181. PubMed DOI PMC

NUNES HP, FURNAS S, DINIS M, BORBA A, Da SILVA JM. Effects of chlorogenic acid on the maturation and fertilisation of bovine oocytes and their embryonic development with a comparative bovine granulosa cell co-culture. Anim Reprod (AR) 2018;14:1057–1061. doi: 10.21451/1984-3143-AR885. DOI

ONISZCZUK A, OLECH M, ONISZCZUK T, WOJTUNIK-KULESZA K, WÓJTOWICZ A. Extraction methods, LC-ESI-MS/MS analysis of phenolic compounds and antiradical properties of functional food enriched with elderberry flowers or fruits. Arab J Chem. 2019;12:4719–4730. doi: 10.1016/j.arabjc.2016.09.003. DOI

PATISAUL HB, JEFFERSON W. The pros and cons of phytoestrogens. Front Neuroendocrinol. 2010;31:400–419. doi: 10.1016/j.yfrne.2010.03.003. PubMed DOI PMC

RAINEY WH, SAWETAWAN C, SHAY JW, MICHAEL MD, MATHIS JM, KUTTEH W, BYRD W, CARR BR. Transformation of human granulosa cells with the e6 and e7 regions of human papillomavirus. J Clin Endocrinol Metab. 1994;78:705–710. doi: 10.1210/jcem.78.3.8126145. PubMed DOI

MARISA RIBEIRO A, ESTEVINHO BN, ROCHA F. Microencapsulation of polyphenols - The specific case of the microencapsulation of Sambucus Nigra L. extracts - A review. Trends Food Sci Tech. 2020;105:454–467. doi: 10.1016/j.tifs.2019.03.011. DOI

ROYCHOUDHURY S, HALENAR M, MICHALCOVA K, NATH S, KACANIOVA M, KOLESAROVA A. Green tea extract affects porcine ovarian cell apoptosis. Reprod Biol. 2018;18:94–98. doi: 10.1016/j.repbio.2018.01.007. PubMed DOI

SARAVI SSS, SHOKRZADEH M, SHIRAZI FH. Cytotoxicity of Sambucus ebulus on cancer cell lines and protective effects of vitamins C and E against its cytotoxicity on normal cell lines. Afr J Biotechnol. 2013;12:3360–3365. doi: 10.5897/AJB09.1577. DOI

SCHRODER L, RICHTER DU, PIECHULLA B, CHROBAK M, KUHN C, SCHULZE S, ABARZUA S, JESCHKE U, WEISSENBACHER T. Effects of phytoestrogen extracts isolated from elder flower on hormone production and receptor expression of trophoblast tumor cells JEG-3 and BeWo, as well as MCF7 breast cancer cells. Nutrients. 2016;8:616. doi: 10.3390/nu8100616. PubMed DOI PMC

SIDOR A, GRAMZA-MICHAŁOWSKA A. Advanced research on the antioxidant and health benefit of elderberry (Sambucus nigra) in food - a review. J Funct Foods. 2015;18:941–958. doi: 10.1016/j.jff.2014.07.012. DOI

SIROTKIN A, ZAHORANSKA Z, TARKO A, FABOVA Z, ALWASEL S, HARRATH AH. Plant polyphenols can directly affect ovarian cell functions and modify toluene effects. J Anim Physiol Anim Nutr. 2020;105:80–89. doi: 10.1111/jpn.13461. PubMed DOI

SIROTKIN AV, PELLEOVA B, FABOVA Z, MAKOVICKY P, ALWASEL S, HARRATH AH. Rutin directly affects stimulatory action of FSH on the ovarian cell. Pharma Nutrition. 2021;15:100247. doi: 10.1016/j.phanu.2020.100247. DOI

WOCŁAWEK-POTOCKA I, MANNELLI C, BORUSZEWSKA D, KOWALCZYK-ZIEBA I, WAŚNIEWSKI T, SKARŻYŃSKI DJ. Diverse effects of phytoestrogens on the reproductive performance: Cow as a model. Int J Endocrinol. 2013;2013:650984. doi: 10.1155/2013/650984. PubMed DOI PMC