1. Harley RM, Brighton CA. Chromosome number in the genus Mentha L. Bot J Linn Soc. 1977;74:71–96.
2. Chambers H. Mentha: Genetic resources and the collection at USDA-ARSNCGR-Corvallis. Lam Newsl. 1992;1:3–4.
3. Stamenkovic V. Revised and Expanded. 2nd ed. Leskovac: NIGP Trend; 2005. Our Non-Harming Medicinal Herbs.
4. Naghibi F, Mosaddegh M, Motamed SM, Ghorbani A. Labiatae family in folk medicine in Iran: From ethnobotany to pharmacology. Iran J Pharm Res. 2005;4:63–79.
5. Mkaddem M, Bouajila J, Ennajar M, Lebrihi A, Mathieu F, Romdhane M. Chemical composition and antimicrobial and antioxidant activities of Mentha (longifolia L. and viridis) essential oils. J Food Sci. 2009;74:M358–63.[PubMed]
6. Mimica-Dukic N, Bozin B, Sokovic M, Mihajlovic B, Matavulj M. Antimicrobial and antioxidant activities of three Mentha species essential oils. Planta Med. 2003;69:413–9.[PubMed]
7. Gulluce M, Sahin F, Sokmen M, Ozer H, Daferera D, Sokmen A, et al. Antimicrobial and antioxidant properties of the essential oils and methanol extract from Mentha longifolia L. ssp. longifolia. Food Chem. 2007;103:1449–56.
8. Hajlaoui H, Snoussi M, Ben Jannet H, Mighri Z, Bakhrouf A. Comparison of chemical composition and antimicrobial activities of Mentha longifolia L. ssp. longifolia essential oil from two Tunisian localities (Gabes and Sidi Bouzid) Ann Microbiol. 2008;58:513–20.
9. Al-Bayati FA. Isolation and identification of antimicrobial compound from Mentha longifolia L. leaves grown wild in Iraq. Ann Clin Microbiol Antimicrob. 2009;8:20.[PMC free article][PubMed]
10. Hussain AI, Anwar F, Nigam PS, Ashraf M, Gilani AH. Seasonal variation in content, chemical composition and antimicrobial and cytotoxic activities of essential oils from four Mentha species. J Sci Food Agric. 2010;90:1827–36.[PubMed]
11. Karaman I, Sahin F, Güllüce M, Ogütçü H, Sengül M, Adigüzel A. Antimicrobial activity of aqueous and methanol extracts of Juniperus oxycedrus L. J Ethnopharmacol. 2003;85:231–5.[PubMed]
12. Kitic D, Jovanovic T, Ristic M, Palic R, Stojanovic G. Chemical composition and antimicrobial activity of the essential oil of Calamintha nepeta (L.) Savi ssp. glandulosa (Req.) P.W. Ball from Montenegro. J Essent Oil Res. 2002;14:150–2.
13. Sahin F, Karaman I, Güllüce M, Ogütçü H, Sengül M, Adigüzel A, et al. Evaluation of antimicrobial activities of Satureja hortensis L. J Ethnopharmacol. 2003;87:61–5.[PubMed]
14. Hafedh H, Fethi BA, Mejdi S, Emira N, Amina B. Effect of Mentha longifolia L. ssp longifolia essential oil on the morphology of four pathogenic bacteria visualized by atomic force microscopy. Afr J Microbiol Res. 2010;4:1122–7.
15. Hajlaoui H, Trabelsi N, Noumi E, Snoussi M, Fallah H, Ksouri R, et al. Biological activities of the essential oils and methanol extract of tow cultivated mint species (Mentha longifolia and Mentha pulegium) used in the Tunisian folkloric medicine. World J Microbiol Biotechnol. 2009;25:2227–38.
16. Khattak S, Rehman SU, Khan T, Shah HU, Shad AA, Ahmad M. In vitro screening for biological pharmacological effects of indigenous medicinal plants, Mentha longifolia and Aloe vera. J Chem Soc Pak. 2004;26:248–51.
17. Gibriel YA, Hamza AS, Gibriel AY, Mohsen SM. In vivo effect of mint (Mentha viridis) essential oil on growth and aflatoxin production by Aspergillus flavus isolated from stored corn. J Food Saf. 2011;31:445–51.
18. El-Badry AA, Al-Ali KH, El-Badry YA. Activity of Mentha longifolia and Ocimum basilicum against Entamoeba histolytica and Giardia duodenalis. Sci Parasitol. 2010;11:109–17.
19. Mann CM, Cox SD, Markham JL. The outer membrane of Pseudomonas aeruginosa NCTC 6749 contributes to its tolerance to the essential oil of Melaleuca alternifolia (tea tree oil) Lett Appl Microbiol. 2000;30:294–7.[PubMed]
20. Cosentino S, Tuberoso CI, Pisano B, Satta M, Mascia V, Arzedi E, et al. In-vitro antimicrobial activity and chemical composition of sardinian thymus essential oils. Lett Appl Microbiol. 1999;29:130–5.[PubMed]
21. Al-Younis NK, Argushy ZM. Antibacterial evaluation of some medicinal plants from Kurdistan region. J Duhok Univ. 2009;12:256–61.
22. Akroum S, Bendjeddou D, Dand S, Lalaoui K. Antibacterial activity and acute toxicity effect of flavonoids extracted from Mentha longifolia. Am Eurasian J Sci Res. 2009;4:93–6.
23. Matsuda H, Morikawa T, Ando S, Toguchida I, Yoshikawa M. Structural requirements of flavonoids for nitric oxide production inhibitory activity and mechanism of action. Bioorg Med Chem. 2003;11:1995–2000.[PubMed]
24. Miyazawa M, Hisama M. Antimutagenic activity of flavonoids from Chrysanthemum morifolium. Biosci Biotechnol Biochem. 2003;67:2091–9.[PubMed]
25. Xiao M, Shao YD, Yan WD, Zhang ZZ. Measurement and correlation of solubilities of apigenin and apigenin7-O-rhamnosylglucoside in seven solvents at different temperatures. J Chem Thermodyn. 2011;43:240–3.
26. Sarac N, Ugur A. Antimicrobial activities and usage in folkloric medicine of some Lamiaceae species growing in Mugla, Turkey. Eurasian J Bio Sci. 2007;4:28–37.
27. Yigit D, Yigit N, Ozgen U. An investigation on the anticandidal activity of some traditional medicinal plants in Turkey. Mycoses. 2009;52:135–40.[PubMed]
28. Gursoy N, Sihoglu-Tepe A, Tepe B. Determination of in vitro antioxidative and antimicrobial properties and total phenolic contents of Ziziphora clinopodioides, Cyclotrichium niveum, and Mentha longifolia ssp. typhoides var. typhoides. J Med Food. 2009;12:684–9.[PubMed]
29. Snoussi M, Hajlaoui H, Noumi E, Usai D, Sechi LA, Zanetti S, Bakhrouf A. In vitro anti-Vibrio spp. activity and chemical composition of some Tunisian aromatic plants. World J Microbiol Biotechnol. 2008;24:3071–6.
30. Shahverdi AR, Rafii F, Tavassoli F, Bagheri M, Attar F, Ghahraman A. Piperitone from Mentha longifolia var. chorodictya Rech F. reduces the nitrofurantoin resistance of strains of enterobacteriaceae. Phytother Res. 2004;18:911–4.[PubMed]
31. Lang G, Buchbauer G. A review on recent research results (2008-2010) on essential oils as antimicrobials and antifungals. A review. Flavour Fragr J. 2011;27:13–39.
32. Pajohi MR, Tajik H, Farshid AA, Basti AA, Hadian M. Effects of Mentha longifolia L. essential oil and nisin alone and in combination on Bacillus cereus and Bacillus subtilis in a food model and bacterial ultrastructural changes. Foodborne Pathog Dis. 2011;8:283–90.[PubMed]
33. Kozan E, Küpeli E, Yesilada E. Evaluation of some plants used in Turkish folk medicine against parasitic infections for their in vivo anthelmintic activity. J Ethnopharmacol. 2006;108:211–6.[PubMed]
34. Abou-Jawdah Y, Sobh H, Salameh A. Antimycotic activities of selected plant flora, growing wild in Lebanon, against phytopathogenic fungi. J Agric Food Chem. 2002;50:3208–13.[PubMed]
35. Cordero C, Zebelo SA, Gnavi G, Griglione A, Bicchi C, Maffei ME, et al. HS-SPME-GC×GC-qMS volatile metabolite profiling of Chrysolina herbacea frass and Mentha spp. leaves. Anal Bioanal Chem. 2012;402:1941–52.[PubMed]
36. Odeyemi OO, Masika P, Afolayan AJ. Insecticidal activities of essential oil from the leaves of Mentha longifolia L. subsp. capensis against Sitophilus zeamais (Motschulsky) (Coleoptera: Curculionidae) Afr Entomol. 2008;16:220–5.
37. Kumar A, Shukla R, Singh P, Singh AK, Dubey NK. Use of essential oil from Mentha arvensis L. to control storage moulds and insects in stored chickpea. J Sci Food Agric. 2009;89:2643–9.
38. Pascual-Villalobos MJ, Robledo A. Screening for anti-insect activity in Mediterranean plants. Ind Crops Prod. 1998;8:183–94.
39. Cetin H, Cinbilgel I, Yanikoglu A, Gokceoglu M. Larvicidal activity of some Labiatae (Lamiaceae) plant extracts from Turkey. Phytother Res. 2006;20:1088–90.[PubMed]
40. Saljoqi AU, Afridi MK, Khan SA, Rehman S. Effects of six plant extracts on rice weevil Sitophilus oryzae L. in the stored wheat grains. J Agric Biol Sci. 2006;1:1–5.
41. Amabeoku GJ, Erasmus SJ, Ojewole JA, Mukinda JT. Antipyretic and antinociceptive properties of Mentha longifolia Huds. (Lamiaceae) leaf aqueous extract in rats and mice. Methods Find Exp Clin Pharmacol. 2009;31:645–9.[PubMed]
42. López V, Martín S, Gómez-Serranillos MP, Carretero ME, Jäger AK, Calvo MI. Neuroprotective and neurochemical properties of mint extracts. Phytother Res. 2010;24:869–74.[PubMed]
43. Pérez Raya MD, Utrilla MP, Navarro MC, Jiménez J. CNS activity of Mentha rotundifolia and Mentha longifolia essential oil in mice and rats. Phytother Res. 2006;4:232–4.
44. Mimica-Dukiç N, Jakovljeviç V, Mira P, Gasiç O, Zabo A. Pharmacological study of Mentha longifolia phenolic extracts. Pharm Biol. 1996;34:359–64.
45. Jan S, Khan MA, Uddin S, Murad W, Hussain M, Ghani A. Herbal recipes used for gastrointestinal disorders in Kaghan valley, nwfp, Pakistan. Pak J Weed Sci Res. 2008;14:169–200.
46. Khan SW, Khatoon S. Ethnobotanical studies on some useful herbs of Haramosh and Bugrote valleys in Gilbit, Northern areas of Pakistan. Pak J Bot. 2008;40:43–58.
47. Hussain K, Shahazad A, Zia-ul-Hussnain S. An ethnobotanical survey of important wild medicinal plants of Hattar District Haripur, Pakistan. Ethnobotanical Lealf. 2008;12:29–35.
48. Cakilcioglu U, Khatun S, Turkoglu I, Hayta S. Ethnopharmacological survey of medicinal plants in Maden (Elazig-Turkey) J Ethnopharmacol. 2011;137:469–86.[PubMed]
49. Shah AJ, Bhulani NN, Khan SH, Ur Rehman N, Gilani AH. Calcium channel blocking activity of Mentha longifolia L. explains its medicinal use in diarrhoea and gut spasm. Phytother Res. 2010;24:1392–7.[PubMed]
50. Naseri MK, Naseri ZG, Mohammadian M, Birgani MO. Ileal relaxation induced by Mentha longifolia (L.) leaf extract in rat. Pak J Biol Sci. 2008;11:1594–9.[PubMed]
51. Jalilzadeh AG, Maham M, Dalir-Naghadeh B, Kheiri F. Effects of Mentha longifolia essential oil on ruminal and abomasal longitudinal smooth muscle in sheep. J Essent Oil Res. 2012;24:61–9.
52. Sousa PJ, Magalhães PJ, Lima CC, Oliveira VS, Leal-Cardoso JH. Effects of piperitenone oxide on the intestinal smooth muscle of the guinea pig. Braz J Med Biol Res. 1997;30:787–91.[PubMed]
53. Stanfill SB, Calafat AM, Brown CR, Polzin GM, Chiang JM, Watson CH, et al. Concentrations of nine alkenylbenzenes, coumarin, piperonal and pulegone in Indian bidi cigarette tobacco. Food Chem Toxicol. 2003;41:303–17.[PubMed]
54. Nagy M, Krizková L, Mucaji P, Kontseková Z, Sersen F, Krajcovic J. Antimutagenic activity and radical scavenging activity of water infusions and phenolics from ligustrum plants leaves. Molecules. 2009;14:509–18.[PubMed]
55. Romanová D, Vachálková A, Cipák L, Ovesná Z, Rauko P. Study of antioxidant effect of apigenin, luteolin and quercetin by DNA protective method. Neoplasma. 2001;48:104–7.[PubMed]
56. Baris O, Karadayi M, Yanmis D, Guvenalp Z, Bal T, Gulluce M. Isolation of 3 flavonoids from Mentha longifolia (L.) Hudson subsp. longifolia and determination of their genotoxic potentials by using the E. coli WP2 test system. J Food Sci. 2011;76:T212–7.[PubMed]
57. Ahmad N, Fazal H, Ahmad I, Abbasi BH. Free radical scavenging (DPPH) potential in nine Mentha species. Toxicol Ind Health. 2012;28:83–9.[PubMed]
58. Berselli PV, Zava S, Montorfano G, Corsetto PA, Krzyzanowska J, Oleszek W, et al. A mint purified extract protects human keratinocytes from short-term, chemically induced oxidative stress. J Agric Food Chem. 2010;58:11428–34.[PubMed]
59. Said O, Saad B, Fulder S, Khalil K, Kassis E. Weight loss in animals and humans treated with “weighlevel”, a combination of four medicinal plants used in traditional arabic and islamic medicine. Evid Based Complement Alternat Med 2011. 2011:874538.[PMC free article][PubMed]
60. Stanisavljevic DM, Stojicevic SS, Ðorcevic SM, Zlatkovic BP, Veliĉkovic DT, Karabegovic IT, et al. Antioxidant activity, the content of total phenols and flavonoids in the ethanol extracts of Mentha longifolia (l.) Hudson dried by the use of different techniques. Chem Ind Chem Eng Q. 2012;18:411–20.
61. Khan FA, Khan A, Hussain J, Khattak MR, Shah SM, Hassan M. Assessment of antioxidant and antibacterial activities of Mentha longifolia. J Pharm Res. 2011;4:2338–9.
62. Ebrahimzadeh MA, Nabavi SM, Nabavi SF. Antioxidant and antihemolytic activities of Mentha longifolia. Pharmacologyonline. 2010;2:464–71.
63. Raj JX, Bajpjpai PK, Kumar PG, Murugan PM, Kumar J, Chaurasia OP, et al. Determination of total phenols, free radical scavenging and antibacterial activities of Mentha longifolia Linn. Hudson from the cold desert, Ladakh, India. Pharmacogn J. 2010;2:470–5.
64. Dudai N, Segey D, Haykin-Frenkel D, Eshel A. Genetic variation of phenolic compounds content essential oil composition and antioxidative activity in Israel-grown Mentha longifolia L. ISHS Acta Hortic. 2006;709:69–78.
65. Odeyemi OO, Yakubu MT, Masika PJ, Afolayan AJ. Toxicological evaluation of the essential oil from Mentha longifolia L. subsp. capensis leaves in rats. J Med Food. 2009;12:669–74.[PubMed]
66. Okoh OO, Afolayan AJ. The effects of hydrodistillation and solvent free microwave extraction methods on the chemical composition and toxicity of essential oils from the leaves of Mentha longifolia L. subsp. capensis. Afr J Pharm Pharmacol. 2011;5:2474–8.
67. Asekun OT, Grierson DS, Afolayan AJ. Effects of drying methods on the quality and quantity of the essential oil of Mentha longifolia L. subsp. Capensis. Food Chem. 2007;101:995–8.
68. Orhan F, Baris Ö, Yanmis D, Bal T, Güvenalp Z, Güllüce M. Isolation of some luteolin derivatives from Mentha longifolia (L.) Hudson subsp. longifolia and determination of their genotoxic potencies. Food Chem. 2012;135:764–9.[PubMed]
69. Gulluce M, Orhan F, Adiguzel A, Bal T, Guvenalp Z, Dermirezer LO. Determination of antimutagenic properties of apigenin-7-O-rutinoside, a flavonoid isolated from Mentha longifolia (L.) Huds. ssp. longifolia with yeast DEL assay. Toxicol Ind Health. 2013;29:534–40.[PubMed]
70. Gulluce M, Yanmis D, Orhan F, Bal T, Karadayi M, ªahin F. Determination of antimutagenic properties of rosmarinic acid, a phenolic compound isolated from Mentha longifolia ssp. longifolia with yeast DEL assay. Microbes Appl Res. 2011;107:526–30.
71. Tuncturk M, Tuncturk R, Sekeroglu N, Ertus MM, Ozgokce F. Lead concentrations of herbs used in Van Herby cheese. Nat Prod Commun. 2011;6:1473–4.[PubMed]
72. Ehsani A, Mahmoudi R. Effects of Mentha longifolia L. essential oil and Lactobacillus casei on the organoleptic properties and on the growth of Staphylococcus aureus and Listeria monocytogenes during manufacturing, ripening and storage of Iranian white-brined cheese. Int J Dairy Technol. 2012;66:70–6.
73. Mahmoudi R, Tajik H, Ehsani A, Farshid AA, Zare P, Hadian M. Effects of Mentha longifolia L. essential oil on viability and cellular ultrastructure of Lactobacillus casei during ripening of probiotic Feta cheese. Int J Dairy Technol. 2012;66:77–82.
74. Hamayun M, Khan SA, Sohn EY, Lee IJ. Folk medicinal knowledge and conservation status of some economically valued medicinal plants of District Swat, Pakistan. Lyonia. 2006;11:101–13.
75. Dwivedi S, Dwivedi A, Dwivedi SN. Folk lore uses of some plants by the tribes of Madhya Pradesh with special reference to their conservation. Ethnobotanical Lealf. 2008;12:763–71.
76. Pirbalouti AG, Malekpoor F, Enteshari S, Yousefi M, Momtaz H, Hamedi B. Antibacterial activity of some folklore medicinal plants used by Bakhtiari tribal in Southwest Iran. Int J Biol. 2010;2:55–63.
77. Darwish RM, Aburjai TA. Effect of ethnomedicinal plants used in folklore medicine in Jordan as antibiotic resistant inhibitors on Escherichia coli. BMC Complement Altern Med. 2010;10:9.[PMC free article][PubMed]
78. Spiridon I, Bodirlau R, Teaca CA. Total phenolic content and antioxidant activity of plants used in traditional Romanian herbal medicine. Cent Eur J Biol. 2011;6:388–96.
79. Karousou R, Balta M, Hanlidou E, Kokkini S. “Mints”, smells and traditional uses in Thessaloniki (Greece) and other Mediterranean countries. J Ethnopharmacol. 2007;109:248–57.[PubMed]
80. Tuzlacı E, Doğan A. Turkish folk medicinal plants, IX: Ovacık (Tunceli) Marmara Pharm J. 2010;14:136–43.
81. Ozgen U, Kaya Y, Houghton P. Folk medicines in the villages of Ilıca District (Erzurum, Turkey) Turk J Biol. 2012;36:93–106.
82. Agelet A, Vallès J. Studies on pharmaceutical ethnobotany in the region of Pallars (Pyrenees, Catalonia, Iberian Peninsula). Part II. New or very rare uses of previously known medicinal plants. J Ethnopharmacol. 2003;84:211–27.[PubMed]
83. López V, Martín S, Gómez-Serranillos MP, Carretero ME, Jäger AK, Calvo MI. Neuroprotective and neurological properties of Melissa officinalis. Neurochem Res. 2009;34:1955–61.[PubMed]
84. Ghoulami S, Il Idrissi A, Fkih-Tetouani S. Phytochemical study of Mentha longifolia of Morocco. Fitoterapia. 2001;72:596–8.[PubMed]
85. Rasooli I, Rezaei MB. Bioactivity and chemical properties of essential oils from Zataria multiflora Boiss and Mentha longifolia (L.) Huds. J Essent Oil Res. 2002;14:141–6.
86. Santos FA, Rao VS. Antiinflammatory and antinociceptive effects of 1,8-cineole a terpenoid oxide present in many plant essential oils. Phytother Res. 2000;14:240–4.[PubMed]
87. Rice KC, Wilson RS. (-)-3-Isothujone, a small nonnitrogenous molecule with antinociceptive activity in mice. J Med Chem. 1976;19:1054–7.[PubMed]
88. Galeotti N, Di Cesare Mannelli L, Mazzanti G, Bartolini A, Ghelardini C. Menthol: A natural analgesic compound. Neurosci Lett. 2002;322:145–8.[PubMed]
89. Oyedeji OA, Afolayan AJ. Chemical composition and antibacterial activity of the essential oil isolated from South African Mentha longifolia L. subsp. capensis (Thunb.) Briq. J Essent Oil Res. 2006;18:57–9.
90. Kaur C, Kapoor HP. Anti-oxidant activity and total phenolic content of some Asian vegetables. Int J Food Sci Technol. 2002;37:153–63.
91. Daferera DJ, Ziogas BN, Pollissiou MG. The effectiveness of plant essential oils on the growth of botrycinera, Fusarium sp. And Clavibacter michiganensis ssp. Michiganensis. Crop Prot. 2003;22:39–44.
Evaluation of possible toxic effects of spearmint (Mentha spicata) on the reproductive system, fertility and number of offspring in adult male rats
Fatemeh Nozhat,1Sanaz Alaee,2,*Khodabakhsh Behzadi,3 and Najmeh Azadi Chegini4
1Department of Biology, Payame Noor University (PNU), IRAN
2Department of Reproductive Biology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, I. R. Iran
3Department of Biology, Sciences and Research Branch, Islamic Azad University, Tehran, I. R. Iran
4Biology Group, Department of Education, Fars, I. R. Iran
*Corresponding Author: Tel: +989171094051, Fax: +987112304372, email@example.com
Author information ►Article notes ►Copyright and License information ►
Received 2013 Oct 18; Revised 2014 Mar 26; Accepted 2014 Jun 25.
Avicenna J Phytomed. 2014 Nov-Dec; 4(6): 420–429.
This article has been cited by other articles in PMC.
Objective: In this study we investigated the effects of spearmint (Mentha spicata Labiatae) on the reproductive system, fertility and number of offspring in adult male rats.
Materials and Methods: Adult Wistar male rats in one control (C) and three experimental groups (I, II and III) received 0, 10, 20 and 40 mg/kg spearmint extract orally for 45 days, respectively. Following this treatment, the animals’ weights, and the standard weight of reproductive tissues, sperm count, sperm motility and serum testosterone concentration were measured, and reproductive tissues were examined histopathologically. To evaluate the effects of spearmint on fertility of male rats and growth of their offspring, male rats of the control and experimental groups mated with untreated female rats.
Results: Results showed that spearmint did not affect the rats’ body and reproductive tissue weights. The sperm count, fast and slow progressive motility of sperm and serum testosterone concentration decreased while number of non-progressive sperm and immotile sperm increased in the experimental groups compared to the control group, but none of these changes were statistically significant. Histopathological studies showed no severe changes in reproductive tissues between control and experimental groups. Number and growth of offspring born from mating of male rats with untreated female rats showed no difference.
Conclusion: We concluded that spearmint has no significant toxic effect on the reproductive system, fertility and number of offspring in adult male rats at the above mentioned dose levels. However high levels of this extract may have adverse effects on male fertility.
Key Words: Fertility, Male, Spearmint, Sperm, Toxicity
Since Iran has many diverse herbal plants, traditional medicine is widely used for treatment of many diseases in this country. The Lamiaceae family is one of the families of flowering plants (Naghibi et al., 2005 ▶) and genus Mentha, an important member of this family, has 6 species in the flora of Iran. These species have great importance in folk medicine and are available in traditional medicinal plant stores and local markets (Amin, 2005 ▶). Spearmint is one of these species, with a good flavor and fragrance, used worldwide in pharmaceutical preparations, confectionery and food industries, and also in hygiene and cosmetic products (Kumar et al., 2008 ▶; Spirling and Daniels, 2001 ▶). Some studies have been shown that spearmint oil has anti-fungal, anti-microbial, anti-inflammatory, anti-tumor and antioxidant activity (Guimaraes et al., 2011 ▶; Lixandru et al., 2010 ▶; Mazzio and Soliman, 2009 ▶; Pearson et al., 2012 ▶; Soković et al., 2009 ▶). Furthermore, various beneficial medicinal effects of spearmint have been found, such as preventing chemotherapy-induced nausea and vomiting (CINV), treatment of respiratory and digestive system disorders, hypertension, anxiety and even for relieving menstrual pain (Cakilcioglu et al., 2011 ▶; Gomez Estrada et al., 2011 ▶; Karousou et al., 2007 ▶; Rokaya et al., 2010 ▶; Tayarani-Najaran et al., 2013; Vejdani et al., 2006; Yoney et al., 2010 ▶). Spearmint is mainly recommended for its antispasmodic effects, which are related to its carvone content, the most important constituent of spearmint (29–74%). Spearmint also contains 4–24% limonene, 0.21–2.1% volatile oil and 3–18% cireole (Baser, 1993).
There is some evidence which show that despi te its beneficial effects, spearmint has some toxic and adverse effects. Severe histopathological changes in kidney, liver and uterus tissue (Akdogan et al., 2004a ▶; Akdogan et al., 2003 ▶; Guney et al., 2006 ▶) and also contact allergic reaction to the leaves of spearmint have been reported (Bonamonte et al., 2001). Furthermore, daily consumption of four cups of spearmint tea can diminish libido in men (Akdogan et al., 2007 ▶). In one study, Akdogan showed that spearmint herbal tea has adverse effects on testicular tissue and testosterone level, and alters the level of follicular stimulating hormone (FSH) and luteinizing hormone (LH) (Akdogan et al., 2004b ▶; Kumar et al., 2008 ▶).
Since spearmint is widely used for digestive problems in Iran, in this study we investigated the possible toxic effects of this agent on the male reproductive system by evaluation of reproductive tissues’ histopathology, plasma testosterone concentration, sperm concentration, spermmotility and number of offspring in adult male rats.
Materials and Methods
Preparation of hydroalcoholic extract of spearmint
Fresh spearmint was purchased from a local market source in Shiraz. The plant’s identity was confirmed by a botanist in the Biology Department, Payame Noor University, Shiraz, Iran, and a voucher specimen (1275) was deposited in this Department. Hydroalcoholic extract was prepared using the maceration method. Leaves were cleaned and dried under shade at room temperature. The dried leaves of the plant were powdered (400 g) and macerated in 1400 ml ethanol for 3 days. Then the solution from the total extract was filtered with filter paper, concentrated by evaporation and stored in refrigerator until used for our experiments. The yield (w/w) of the solution was 9.6% (g/g).
Adult Wistar male rats of proven fertility, 8–10 weeks old age and weighing 200–250 g were kept in laboratory conditions for adaptation two weeks before experiments. They were maintained in a well-ventilated animal house under standard conditions and controlled temperature (22–24 °C), with periods of 12 hours light and 12 hours darkness. The animals had sufficient access to food and water throughout the study. The weights of animals were recorded before and after the experiments. The animal experiments were performed according to the principles of the care and use of laboratory animals established by the National Institutes of Health (NIH Publication, 1985). Then male rats were randomly divided into one control (C) and three experimental groups (I, II, III).
There were 15 animals in each group (9 animals for histopathological and sperm parameters evaluation and 6 animals for mating with untreated female rats). Group Ι received 10 mg/kg, group II received 20 mg/kg and group Ш received 40 mg/kg of spearmint leaf extract orally for 45 days. The control group received 1 ml of distilled water daily.
After 45 days, animals were sacrificed using diethyl ether. Blood samples were collected by dorsal aorta puncture, centrifuged at 3000 rpm for 15 min, and serum was separated. The concentration of serum testosterone was assayed by solid phase radioimmunoassay (RIA) method.
Standard weight of reproductive tissues
After scarifying the animals and collecting blood samples, testes, epididymis, seminal vesicles and prostate were removed and weighed. The standard weights of right testis, epididymis, seminal vesicle and prostate was calculated by following formula: [tissue weight (g) / body weight (g)] × 100.
The testes, epididymis, seminal vesicles and prostate of each rat were fixed in 10% buffer formalin solution. Tissues were processed for preparation of paraffin blocks, which then were sectioned at a thickness of 6 µm using a microtome, and stained with hematoxylin and eosin.
Sperm movement and count
During dissection of each rat, the distal portion of each vas deferens (1 cm) was removed and placed in a falcon tube contained 5 ml Hanks’ solution at 37 ºC for capture of sperm exiting from the vas deferens (Seed et al., 1996 ▶). After 3 minutes, one drop of Hanks’ solution containing sperm was placed on a microscope slide and for each animal, movement of 100 spermatozoa were assessed by light microscope at 40x magnification. Sperm movements were divided into 4 grades of fast progressive movement, slow progressive movement, non-progressive movement and immotile sperm, according to the WHO Laboratory Manual for the Examination and Processing of Human Semen (WHO, 2010 ▶). After 10 minutes, sperm count was assessed by putting one drop of sperm suspension on a hemocytometer (Da Silveira et al., 2003 ▶). Total sperm number was calculated using the following formula: A= B×C×D,
A: total sperm number in 1 cm of vas deferens
B: total sperm number in 0.1 mm3 of solution
C: deep factor
D: dilution factor= 5000mm3
To evaluate the effect of spearmint on the fertility of male rats and the growth of their offspring, following the last day of spearmint administration, 6 male rats from each group were cohabitated with proestrus untreated females rats for mating, per the following design:
1) 6 male rats of group C mated with 6 untreated female rats
2) 6 male rats of group I mated with 6 untreated female rats
3) 6 male rats of group II mated with 6 untreated female rats
4) 6 male rats of group III mated with 6 untreated female rats
Vaginal smear was examined every morning to determine positive mating. After completion of the duration of the female rats’ pregnancies, the number, weight and crown-rump length (CRL) of offspring from each pregnant female rat were recorded (Monsefi et al., 2010 ▶).
Statistical analysis was done using SPSS 11.5 software. For data analysis, we used one-way ANOVA followed by the Tukey post hoc test. Results were expressed as mean ± SEM (standard error of the mean) and p<0.05 was considered statistically significant.
Comparison of body weight of rats at the beginning and end of the experiment showed no significant difference between control and experimental groups. The standard weight of right testis, epididymis, seminal vesicle and prostate had no significant change in experimental groups when compared to the control group (Table 1). The serum testosterone concentration, sperm count and the number of sperm with fast and slow progressive movement were lower in experimental groups compared to the control group but not statistically significant (Table 2). The number of non-progressive and immotile sperm was higher in experimental groups compared to the control group, but none of these changes were statistically significant (Table 2).
The standard weight (g) of right testis, seminal vesicle, epididymis and prostate in control, group I, group II and group III male rats. (n=9 for each group)
Serum testosterone concentration (ng/ml), sperm count and sperm motility in control, group I, group II and group III male rats. (n=9 for each group
Histopathological studies showed no change in the structure of reproductive tissues between experimental groups and the control group. All seminiferous tubules of testis tissue had normal histopathological features in control and experimental groups. Spermatogenic cells at all stages of spermatogenesis (spermatogonia, primary spermatocytes, secondary spermatocytes, spermatids and spermatozoids) were seen in these tubules, demonstrating that the normal spermatogenesis process has occurred in these animals (Figure 1). The epididymal tissue in all groups showed normal structure with pseudostratified columnar epithelium and normal sperm density (Figure 2).
The seminiferous tubules in the control group (A) group I (B) group II (C) and group III (D) of male rats. Hematoxylin and eosin staining, 10x magnification
The epididymis tissue in the control group (A) group I (B) group II (C) and group III (D) of male rats. Hematoxylin and eosin staining, 10x magnification
Furthermore, no noticeable histopathological change was observed in seminal vesicle and prostate. The tubules in the seminal vesicle showed normal pseudostratified columnar epithelium. In prostate tissue a normal tubuloalveolar structure with normal epithelium and smooth muscles fibers observed in all groups (Figures 3 and 4).
The seminal vesicle tissue in the control group (A) group I (B) group II (C) and group III (D) of male rats. Hematoxylin and eosin staining, 10x magnification
The prostate tissue in the control group (A) group I (B) group II (C) and group III (D) of male rats. Hematoxylin and eosin staining, 10x magnification
No statistically significant difference was observed in the weight and CRL of offspring and the pregnancy outcome of control and experimental male rats mated with untreated female rats (Table 3).
The number, weight (g) and crown-rump length (cm) of offspring from mating of control, group I, group II and group III male rats, (n=6 for each group) with 24 female untreated rats
Spearmint is a herbal plant which is usually recommended for treatment of many diseases, particularly digestive system problems; however, some studies show that despite its beneficial effects, spearmint has adverse effects on the reproductive system of male rats (Akdogan et al., 2004b ▶; Kumar et al., 2008 ▶). Spearmint reduces free testosterone concentration of serum and has been introduced as an anti-androgenic agent and proposed for treatment of hirsuitism in polycystic ovarian syndrome (PCOS) of women (Akdogan et al., 2007 ▶; Grant, 2010 ▶). Therefore, this study was designed to investigate the effects of spearmint extract on the male reproductive system and fertility outcome.
One of the methods to determine toxicity of plant extracts is measurement of any changes in the body weight (Gupta and Sharma, 2006 ▶). Given no alteration in the body weight of experimental rats in our study, this medicinal plant has no general toxicity effects at the level of administered doses; this circumstance was observed by other studies (Guney et al., 2006 ▶; Kumar et al., 2008 ▶).
For successful fertility, normal structure and accurate function of all parts of reproductive system is needed. A complex mechanism under the regulated function of the hypothalamic-pituitary-gonadal axis (HPG axis) is responsible for initiation and maintenance of spermatogenetic activity. Initially, by secretion of GnRH (gonadotropin-releasing hormone) from the hypothalamus, FSH and LH are released from the pituitary gland. In the testes, under the stimulatory action of LH, the Leydig cells, located in interstitial tissue, produce and secrete testosterone. Simultaneously, FSH supports the function of sertoli cells, a mediator for effects of testosterone and FSH on germ cells for successful spermatogenesis in seminiferous tubules.
After that, produced sperm pass through the epididymis, which secretes substances for sperm maturation (Cooper, 2002 ▶). Ultimately, through the secretion of substances such as fructose, citrate, inositol and prostaglandins from seminal vesicles and secretion of prostatic liquid from the prostate, semen is produced. Thus any pathological changes in male reproductive tissues may interfere with fertility by altering the level of testosterone hormone or disturbing the spermatogenesis and sperm maturation process (Hafez and Hafez, 2005 ▶). In this study, weight and structure of testes did not change in experimental groups compared to the normal group, which resulted in normal testosterone level and also sperm concentration and motility, two critical parameters for male fertility. As epididymis, seminal vesicle and prostate are androgen-dependent tissues, the normal histology of these tissues as seen in control and experimental groups is the consequence of normal concentration of serum testosterone hormone (Nieschlag et al., 2000 ▶).
For evaluation of the fertility outcome of male rats, we also determined the number of their offspring born from mating of animals of all 4 groups with untreated females. Since spearmint administration did not cause any decline in number and motility of sperms, no difference was seen in offspring numbers of experimental male rats compared to control. Moreover, the weight and crown-rump length of offspring were not affected.
Therefore, we concluded that treatment of male rats with the mentioned doses of spearmint extract has no pathological, antiandrogenic or antifertility effects. This result is in contrast with the results of some investigations, which showed that spearmint has an antiandrogenic effect in male rats (Akdogan et al., 2004b ▶; Kumar et al., 2008 ▶). In their studies they used higher doses of spearmint extract and treated animals in a different way. They steeped spearmint tea (dried leaves) in a cup of boiling water and added it to the drinking water. So animals received spearmint continuously at all times for maximum 35 days, while we administered spearmint in lower doses, once a day for a longer time (45 days). Kumar (2008) observed no significant damage to the reproductive system following the short-term use of spearmint, but long-term use caused irreversible damage to this system, such as significant decrease in the weights of seminal vesicle, epididymis, testis and prostate with significant histopathological changes in these tissues. Also the level of LH and FSH decreased, which was attributed to generation of oxidative stress in the hypothalamus and pituitary gland, leading to reduced production of GnRH and gonadotropins and resulting in an attenuated level of testosterone and spermatogenesis arrest in treated rats. In addition, severe histopathologic changes were observed in testicular tissue. Similar results were obtained in the study of Akdogan (2004b) ▶ with the same doses and same duration of experiment. But higher levels of LH and FSH were observed in experimental animals followed by decreases in testosterone level and deficiency of spermatogenesis. They explained that an increase in FSH and LH is a normal process which occurs as the result of a decrease in plasma total testosterone levels and concluded that the deficiency of spermatogenesis is the consequence of the direct effect of spearmint on testicular tissue and Leydig cell dysfunction.
In conclusion, the administration of spearmint at the dosage level used in the present study has no antifertility effect in adult male rats. However, high levels of this extract have adverse effects on male fertility. Therefore, people who consume spearmint should be advised to use this herbal plant in a proper manner and avoid high doses. Furthermore, because the exact constituents of spearmint which cause antifertility have not been clarified, we suggest that ongoing studies should be performed on different fractions of spearmint extract.
This study was financially supported by a research grant from the Vice-Chancellor of the Department of Biology, Payame Noor University, Shiraz, Iran ]grant number 1318.104.22.168.392[.
Conflict of Interest Statement
The authors declare that there are no conflicts of interest.
- Akdogan M, Wnc I, Oncu M, Karaoz E, Delibas NW. Investigation of biochemical and histopathological effects of Mentha piperita L. and Mentha spicata L. on kidney tissue in rats. Hum Exp Toxicol. 2003;22:213–219.[PubMed]
- Akdogan M, Ozguner M, Aydin G, Gokalp O. Investigation of biochemical and histopathological effects of Mentha piperita Labiatae and Mentha spicata Labiatae on liver tissue in rats. Hum Exp Toxicol. 2004a;23:21–28.[PubMed]
- Akdogan M, Ozguner M, Kocak A. Effects of peppermint teas on plasma testosterone, follicle-stimulating hormone and luteinizing hormone levels and testicular tissue in rats. Urology. 2004b;64:394–398.[PubMed]
- Akdogan M, Tamer MN, Cüre E, Cüre MC, Krolu BK, Delibat N. Effect of spearmint (Mentha spicata Labiatae) teas on androgen levels in women with hirsutism. Phytother Res. 2007;21:444–447.[PubMed]
- Amin G. Popular medicinal plants of Iran. Iran: Tehran University of Medical Sciences Press; 2005.
- Cakilcioglu U, Khatun S, Turkoglu I, Hayta S. Ethnopharmacological survey of medicinal plants in Maden (Elazig Turkey) J Ethnopharmacol. 2011;137:469–486.[PubMed]
- Cooper TG. Recent advances in sperm maturation in the human epididymis. Andrologie. 2002;12:38–51.
- Da Silveira RC, Leite MN, Reporedo MM, De Almeida RN. Evaluation of long-term exposure to Mikania glomerata (Sprengel) extract on male Wistar rats’ reproductive organs, sperm production and testosterone level. J Contraception. 2003;67:327–331.[PubMed]
- Gomez Estrada H, Diaz Castillo F, Franco Ospina L, Mercad Camargo J, Guzman Ledezma J, Medina JD, Gaitan Ibarra R. Folk medicine in the northern Coast of Colombia: an overview. J Ethnobiol Ethnomed. 2011;22:7–27.[PMC free article][PubMed]
- Grant P. Spearmint Herbal Tea has Significant Anti-androgen Effects in Polycystic Ovarian Syndrome A Randomized Controlled Trial. . Phytother Res. 2010;24:186–188.[PubMed]
- Guimaraes R, Barreira J C, Barros L, Carvalho AM, Ferreira IC. Effects of Oral Dosage Form and Storage Periodon the Antioxidant Properties of Four Species Used in Traditional Herbal Medicine. Phytother Res. 2011;25:484–492.[PubMed]
- Guney M, Oral B, Karahanl N, Mungana T, Akdogan M. The effect of Mentha spicata Labiatae on uterine tissue in rats. Toxicol Ind Health. 2006;22:343–348.[PubMed]
- Gupta RS, Sharma R. A review on medicinal plants exhibiting antifertility activity in males. Nat Prod Radiance. 2006;5:389–410.
- Hafez ESE, Hafez SD. Atlas of Clinical Andrology. United Kingdom: Taylor & Francis; 2005.
- Karousou R, Balta M, Hanlidou E, Kokkini S. "Mints", smells and traditional uses in Thessaloniki (Greece) and other Mediterranean countries. J Ethnopharmacol. 2007;109:248–257.[PubMed]
- Kumar V, Kural MR, Pereira BMJ, Roy P. Spearmint induced hypothalamic oxidative stress and testicular anti-androgenicity in male rats – altered levels of gene expression, enzymes and hormones. Food Chem Toxicol. 2008;46:3563–3570.[PubMed]
- Lixandru BE, Drăcea NO, Dragomirescu CC, Drăgulescu EC, Coldea IL, Anton L, Dobre E, Rovinaru C, Codiţă I. Antimicrobial activity of plant essential oils against bacterial and fungal species involved in food poisoning and/or food decay. Roum Arch Microbiol Immunol. 2010;69:224–230.[PubMed]
- Mazzio EA, Soliman KF. In Vitro Screening for the Tumoricidal Properties of International Medicinal Herbs. Phytother Res. 2009;23:385–398.[PMC free article][PubMed]
- Monsefi M, Alaee S, Moradshahi A, Rohani L. Cadmium‐induced infertility in male mice. Environ Toxicol. 2010;25:94–102.[PubMed]
- Naghibi F, Mosaddegh M, Mohammadi Motamed S, Ghorbani A. Labiatae Family in folk Medicine in Iran From Ethnobotany to Pharmacology. Iran J Pharm Res. 2005;2:63–79.
- NIH Publication revised. Anonymous Principles of National Institute of Health. 1985. No. 85–2.
- Nieschlag SMA, Nieschlag E, Behre H. Andrology: Male Reproductive Health and DysfunctionHeidelberg. 2nd Edition. Berlin : Spring Springer-Verlag; 2000.
- Pearson W, Fletcher RS, Kott LS. Oral rosmarinic Acid-enhanced Mentha spicata modulates synovial fluid biomarkers of inflammation in horses challenged with intra-articular LPS. J Vet Pharmacol Ther. 2012;35:495–502.[PubMed]
- Rokaya MB, Munzbergova Z, Timsina B. Ethnobotanical study of medicinal plants from The Humla district of western Nepal. J Ethnopharmacol. 2010;130:485–504.[PubMed]
- Seed J, Chapin RE, Clegg ED, Dostal LA, Foote RH, Hurtt ME. Methods for assessing sperm motility, morphology and counts in the rat, rabbit & dog: A consensus report. Reprod Toxicol. 1996;10:237–244.[PubMed]
- Soković MD, Vukojević J, Marin PD, Brkić DD, Vajs V, van Griensven LJ. Chemical Composition of essential oils of Thymus and Mentha species and their antifungal activities. Molecules. 2009;14:238–249.[PubMed]
- Spirling LI, Daniels IR. Botanical perspectives on health peppermint: more than just an after-dinner mint. J R Soc Health. 2001;121:62–63.[PubMed]
- Tayarani-Najaran T, Talasaz-Firoozi E, Nasiri R, Jalali N MK. Antiemetic activity of volatile oil from Mentha spicata and Mentha piperita in chemotherapy-induced nausea and vomiting. Ecancermedicalscience. 2013;7:290.[PMC free article][PubMed]
- Yoney A, Prieto JM, Lardos A, Heinrich M. Ethnopharmacy of Turkish-speaking Cypriots in Greater London. Phytother Res. 2010;24:731–740.[PubMed]
- Vejdani R, Shalmani HR, Mir-Fattahi M, Sajed-Nia F, Abdollahi M, Zali MR, Alizadeh AH, Bahari A, Amin G. The efficacy of an herbal medicine, Carmint, on the relief of abdominal pain and bloating in patients with irritable bowel syndrome: a pilot study. Dig Dis Sci. 2006;51:1501–1507.[PubMed]
- World Health Organization. WHO laboratory manual for the Examination and processing of human semen. Fifth Edition. Switzerland: WHO Press; 2010.
Articles from Avicenna Journal of Phytomedicine are provided here courtesy of Mashhad University of Medical Sciences