Several plants have the potential to protect essential reproductive processes such as spermatogenesis or steroidogenesis, however, effective concentrations and main mechanisms of action are still unknown. This in vitro study was aimed to assess the effects of Apium graveolens L., Levisticum officinale, and Calendula officinalis L. extracts on the structural integrity, functional activity and gap junctional intercellular communication (GJIC) in mice Leydig cells. TM3 cells were grown in the presence of experimental extracts (37.5; 75; 150 and 300 µg/ml) for 24 h. For the present study, high-performance liquid chromatography analysis was used to quantify flavonoids or phenolic acids. Subsequently, Leydig cell viability was assessed by alamarBlue assay, while the cell membrane integrity was detected by 5-carboxyfluorescein diacetate-acetoxymethyl ester. The level of steroid hormones production was determined by enzyme-linked immunosorbent assay. Additionally, GJIC was assessed by scalpel loading/dye transfer assay. According to our results, Apium graveolens L. significantly increased the viability and cell membrane integrity at 75 µg/ml (109.0±4.3%) followed by a decline at 300 µg/ml (89.4±2.3%). In case of Levisticum officinale and Calendula officinalis L. was observed significant decrease at 150 µg/ml (88.8±11.66%; 87.4±6.0%) and 300 µg/ml (86.2±9.3%; 84.1±4.6%). Furthermore, Apium graveolens L. significantly increased the progesterone and testosterone production (75 and 150 µg/ml) however, Levisticum officinale and Calendula officinalis L. significantly reduced steroid hormones synthesis at 150 and 300 µg/ml. Finally, the disturbance of GJIC was significantly affected at 300 µg/ml of Levisticum officinale (82.5±7.7%) and Calendula officinalis L. (79.8±7.0%). The balanced concentration ratio may support the Leydig cell function, steroidogenesis as well as all essential parameters that may significantly improve reproductive functions.

BioFood Centre Faculty of Biotechnology and Food Sciences Slovak University of Agriculture in Nitra Nitra Slovak Republic

Zobrazit více v PubMed

ABBAS MA. Is the use of plants in Jordanian folk medicine for the treatment of male sexual dysfunction scientifically based? Review of in vitro and in vivo human and animal studies. Andrologia. 2017;49:1–21. doi: 10.1111/and.12619. PubMed DOI

ALNUQAYDAN AM, LENEHAN CE, HUGHES RR, SANDERSON BJ. Extracts from Calendula officinalis offer in vitro protection agains H2O2 induced oxidative stress cell killing of human skin cells. Phytother Res. 2015;29:120–124. doi: 10.1002/ptr.5236. PubMed DOI

BENKO F, PALKOVIČOVÁ V, ĎURAČKA M, ÁRVAY J, LUKÁČ N, TVRDÁ E. Antioxidant effects of marigold (Calendula officinalis) flower extract on the oxidative balance of bovine spermatozoa. Contemporary Agriculture. 2019;68:92–102. doi: 10.2478/contagri-2019-0015. DOI

EKIERT H. Medicinal plant biotechnology: the Apiaceae family as the example of rapid development. Pharmazie. 2000;55:561–567. PubMed

FRUM A. HPLC determination of polyphenols from Calendula officinalis L. flowers. AUCFT. 2017;21:97–101. doi: 10.1515/aucft-2017-0020. DOI

GAO LL, ZHOU CHX, SONG XF, FAN KW, LI FR. Inhibition effects of celery seed extract on human stomach cancer cell lines Hs746T. In: Li S, Jin Q, Jiang X, Park JJ, editors. Frontier and Future Development of Information Technology in Medicine and Education. Springer; Switzerland AG: 2014. pp. 2553–2560. DOI

GHAEDI N, POURABOLI I, DABIRI S. Effect of Levisticum officinale extract on oxidative stress markers, testosterone level and histology of testis tissue in diabetic rats. Ir J Physiol Pharmacol. 2018;2:192–200.

GILLERON J. Connexins as potential therapeutic targets for testis pathologies. Cell Mol Med. 2015;1:1–3. doi: 10.21767/2573-5365.100001. DOI

HALO M, JR, MASSANYI P, GREN A, LASAK A, SLANINA T, ONDRUSKA L, MUCHACKA R, GALBAVY D, IVANIC P, SCHNEIR R, FORMICKI G. Time and dose-dependent effect of Viscum album quercus on rabbit spermatozoa motility and viability in vitro. Physiol Res. 2019;68:955–972. doi: 10.33549/physiolres.934223. PubMed DOI

HAMID R, ROTSHTEYN Y, RABADI L, PARIKH R, BULLOCK P. Comparison of alamar blue and MTT assays for high through-put screening. Toxicol In Vitro. 2004;18:703–710. doi: 10.1016/j.tiv.2004.03.012. PubMed DOI

HASAN A, SHAHRAKI A, SHAHRAKI J. The hepatoprotective effects of aquatic extract of Levisticum officinale against paraquat hepatocyte toxicity. Pak J Pharm Sci. 2017;30:2363–2368. PubMed

HELAL MAM. Celary oil modualtes DEHP-induced treproductive toxicity in male rats. Reprod Biol. 2014;14:182–189. doi: 10.1016/j.repbio.2014.04.002. PubMed DOI

JAMBOR T, KOVACIKOVA E, GREIFOVA H, KOVACIK A, LIBOVA L, LUKAC N. Assesment of the effective impact of bishpenols on mitochondrial activity and steroidogenesis in a dose-dependency in mice TM3 Leydig cells. Physiol Res. 2019;68:689–693. doi: 10.33549/physiolres.934200. PubMed DOI

JAMBOR T, ARVAY J, IVANISOVA E, TVRDA E, KOVACIK A, GREIFOVA H, LUKAC N. Investigation of the properties and effects of Salvia Officinalis L. on the viability, steroidogenesis and reactive oxygen species production in TM3 Leydig cells in vitro. Physiol Res. 2020;69:661–673. doi: 10.33549/physiolres.934457. PubMed DOI PMC

KOOTI W, FAROKHIPOUR M, ASADZADEH Z, ASHTARY-LARKY D, ASADI-SAMANI M. The role of medicinal plants in the treatment of diabetes: a systematic review. Electron Physician. 2016;8:1832–1842. doi: 10.19082/1832. PubMed DOI PMC

KUSHWAHA S, AGARWAL M, MUTREJA A, CHAUHAN A. Impact of 50 % ethanolic extract of Calendula officinalis (flower) on the reproductive funcion of male albino rats (Rattus norvegicus) Egypt J Biol. 2007;9:1–5. doi: 10.4314/ejb.v9i1.56552. DOI

LIU J, LIANG P, YIN C, WANG T, LI H, LI Y, YE Z. Effects of several Chinese herbal aqueous extracts on human sperm motility in vitro. Andrologia. 2004;36:78–83. doi: 10.1111/j.1439-0272.2004.00607.x. PubMed DOI

LUKŠIČ L, ÁRVAY J, VOLLMANNOVÁ A, TÓTH T, ŠKRABANJA V, TRČEK J, GERM M, KREFT I. Hydrothermal treatment of Tartary buckwheat grain hinders the transformation of rutin to quercetin. J Cer Sci. 2016;72:131–134. doi: 10.1016/j.jcs.2016.10.009. DOI

MADKOUR NK. Beneficial role of celery oil in lowering the di(2-ethylhexyl) phtalata-induced testicular damage. Toxicol Ind Health. 2014;30:861–872. doi: 10.1177/0748233712464808. PubMed DOI

NAKAMURA Y, CHANG CH, MORI T, SATO K, OHTSUKI K, UPHAM BL, TROSKO JE. Augmentation of differentiation of gap juncion function by kaempferol in partially differentiated colon cancer cells. Carcinogenesis. 2005;26:665–671. doi: 10.1093/carcin/bgi003. PubMed DOI

NANTIA EA, MOUNDIPA PF, MONSEES TK, CARREAU S. Medicinal plants as potential male anti-invertility agents: a review. Bas Clin Adrol. 2009;19:148–158. doi: 10.1007/s12610-009-0030-2. DOI

NOUR V, TRANDAFIR I, COSMELESCU S. Bioactive compounds, antioxidat activity and nutritional quality of different culinar aromatic herbs. Not Bot Horti Agro Cluj Nap. 2017;45:179–184. doi: 10.15835/nbha45110678. DOI

PREETHI KC, SIVEEN KS, KUTTAN R, KUTTAN G. Inhibition of metastasis of B16F-10 melanoma ce BL/6 mice and extract of calendula officinalis L flowers. Asian Pac J Cancer Prev. 2010;11:1773–1779. PubMed

SAHA R, ROYCHODHURY S, KAR K, VARGHESE AC, NANDI P, SHARMA GD, FORMICKI G, SLAMA P, KOLESAROVA A. Coenzyme Q10 ameliorates cadmium induced reproductive toxicity in male rats. Physiol Res. 2019;68:141–145. doi: 10.33549/physiolres.934000. PubMed DOI

SEDEH MA, SIAHPOUSH EA, DARVISHZADEH Z. The investigation of fertility increase and effective factors on it among the kord clan in Andimeshk. JISDS. 2012;4:81–98.

SERTEL S, EICHHORM T, PLINKERT PK, EFFERTH T. Chemical composition and antiproliferative activity of essential oil from the leaves of a medicinal herb, Levisticum officinale, against UMSCC1 head and neck squamous carcinoma cells. Anticancer Res. 2011;32:185–191. PubMed

SCHNEIDER CA, RASBAND WS, ELICEIRI KW. NIH image to ImageJ: 25 years of image analysis. Nat Methods. 2012;9:671–675. doi: 10.1038/nmeth.2089. PubMed DOI PMC

SCHREER A, TINSON CH, SHERRY JP, SCHIRMER K. Application of alamar blue/5-carboxylfluorescein diacetate acetoxymethly ester as a noninvasive cell viability assay in primary hepatocytes from rainbow trout. Anal Biochem. 2005;344:76–85. doi: 10.1016/j.ab.2005.06.009. PubMed DOI

SMITH S. Drubs that cause sexual dysfuncion. Psychiatry. 2007;6:111–114. doi: 10.1016/j.mppsy.2006.12.004. DOI

SUBHADRADEVI V, KHAIRUNISSA K, ASOKKUMAR K, SIVASHANMUGAM MUA, JAGANNAT P. Induction of apoptosis and cytotoxic activities of Apium graveolnes Linn. using in vitro models. Mid East J Sci Res. 2011;9:90–94.

SUN J, WANG H, LIU B, SHI W, SHI J, ZHANG Z, XING J. Rutin attenuates H2O2-iduced oxidation damage and apoptosis in Leydig cells by activating PI3K/Akt signal pathways. Biomed Pharm. 2017;88:500–506. doi: 10.1016/j.biopha.2017.01.066. PubMed DOI

TVRDÁ E, MICHALKO J, MATUŠÍKOVÁ I, LUKÁČ N. In vitro effects of the Chlamydomonas reinhardtii extract on bovine spermatozoa. J Microbiol Biotech Food Sci. 2016;6:972–975. doi: 10.15414/jmbfs.2016/17.6.3.972-975. DOI

TVRDÁ E, VARGA A, SLÁVIK M, ÁRVAY J. Levisticum officinale and its effects on bovine spermatozoa activity. J Microbiol Biotech Food Sci. 2019;8:1212–1216. doi: 10.15414/jmbfs.2019.8.5.1212-1216. DOI

UPHAM BL, SOVADINOVA I, BABICA P. Gap juncional intercellualr communication: a funcional biomarker to assess adverse effects of toxicants and toxins, and health benefits of natural product. J Vis Exp. 2016;118:54281. doi: 10.3791/54281. PubMed DOI PMC

YAO Y, SANG W, ZHOU M, REN G. Phenolic composition and antioxidant activities of 11 celery cultivars. J Food Sci. 2010;75:9–13. doi: 10.1111/j.1750-3841.2009.01392.x. PubMed DOI