Medicinal Chemistry Research

, Volume 24, Issue 11, pp 3876–3884

β-Caryophyllene as putative male contraceptive: enhances spermatogenesis but not spermiogenesis in albino rats

  • Zina M. Al-Alami
  • Ziad A. Shraideh
  • Mutasem O. Taha
Original Research


β-Caryophyllene (β-Car) is a natural antioxidant. It is Cannabis-derived cannabinoid receptors type 2 (CB2R) agonist. CBR2 are expressed in germ cells within the testes. The aim of this study was to investigate the effect of β-Car on the male rat body weight, sex organ weight, sperm count, motility and morphology, and on the histology and the ultrastructure of the testes and the cauda epididymides. Moreover, we compared these effects with the results of the reversible infertility inducer: metronidazole (MTZ). Sexually mature male rats were divided into five groups that received intraperitoneal injections of 10 mg/kg β-Car, 40 mg/kg MTZ, distilled water, tween 80 in normal saline solution (10 % v/v), or did not receive any treatment for an experimental period of 30 days. Cauda epididymal sperm suspensions were used to assess the sperm count, motility, and morphology. Histological and ultrastructural studies were performed on testes and cauda epididymides. Treatment with β-Car (10 mg/kg, i.p.) significantly increased the final body weight, but did not affect the sex organ weight. It resulted in a significant decrease in cauda epididymal sperm count, percentages of motile sperms, and sperms with normal morphology. Nevertheless, it neither affected the histology nor the ultrastructure of the testes or the cauda epididymides of the treated rats. As a CB2R, β-Car could be a promising natural male contraceptive, because 30-day treatment with 10 mg/kg β-Car did not affect spermatogenesis but negatively affected spermiogenesis.


β-Caryophyllene Male fertility Metronidazole Spermatogenesis Spermiogenesis 


  1. Abbas MA, Taha MO, Zihlif MA, Disi AM (2013) Beta-caryophyllene causes regression of endometrial implants in a rat model of endometriosis without affecting fertility. Eur J Pharmacol 702:12–19CrossRefPubMedGoogle Scholar
  2. Agirregoitia E, Carracedo A, Subiran N, Valdivia A, Agirregoitia N, Peralta L, Velasco G, Irazusta J (2010) The cb(2) cannabinoid receptor regulates human sperm cell motility. Fertil Steril 93:1378–1387CrossRefPubMedGoogle Scholar
  3. Amiel E, Ofir R, Dudai N, Soloway E, Rabinsky T, Rachmilevitch S (2012) Beta-caryophyllene, a compound isolated from the biblical balm of gilead (Commiphora gileadensis), is a selective apoptosis inducer for tumor cell lines. Evid Based Complement Alternat Med 2012:1–8CrossRefGoogle Scholar
  4. Baltus AE, Menke DB, Hu YC, Goodheart ML, Carpenter AE, de Rooij DG, Page DC (2006) In germ cells of mouse embryonic ovaries, the decision to enter meiosis precedes premeiotic DNA replication. Nat Genet 38:1430–1434CrossRefPubMedGoogle Scholar
  5. Calleja MA, Vieites JM, Montero-Melendez T, Torres MI, Faus MJ, Gil A, Suarez A (2013) The antioxidant effect of beta-caryophyllene protects rat liver from carbon tetrachloride-induced fibrosis by inhibiting hepatic stellate cell activation. Br J Nutr 109:394–401CrossRefPubMedGoogle Scholar
  6. Chang HJ, Kim JM, Lee JC, Kim WK, Chun HS (2013) Protective effect of beta-caryophyllene, a natural bicyclic sesquiterpene, against cerebral ischemic injury. J Med Food 16:471–480CrossRefPubMedGoogle Scholar
  7. Creasy DM (2003) Evaluation of testicular toxicology: a synopsis and discussion of the recommendations proposed by the society of toxicologic pathology. Birth Defects Res B Dev Reprod Toxicol 68:408–415CrossRefPubMedGoogle Scholar
  8. Di Sotto A, Maffei F, Hrelia P, Castelli F, Sarpietro MG, Mazzanti G (2013) Genotoxicity assessment of beta-caryophyllene oxide. Regul Toxicol Pharmacol 66:264–268CrossRefPubMedGoogle Scholar
  9. El-Nahas AF, El-Ashmawy IM (2004) Reproductive and cytogenetic toxicity of metronidazole in male mice. Basic Clin Pharmacol Toxicol 94:226–231CrossRefPubMedGoogle Scholar
  10. Galdino PM, Nascimento MV, Florentino IF, Lino RC, Fajemiroye JO, Chaibub BA, de Paula JR, de Lima TC, Costa EA (2012) The anxiolytic-like effect of an essential oil derived from spiranthera odoratissima a. St. Hil. Leaves and its major component, beta-caryophyllene, in male mice. Prog Neuropsychopharmacol Biol Psychiatry 38:276–284CrossRefPubMedGoogle Scholar
  11. Gertsch J, Leonti M, Raduner S, Racz I, Chen JZ, Xie XQ, Altmann KH, Karsak M, Zimmer A (2008) Beta-caryophyllene is a dietary cannabinoid. Proc Natl Acad Sci USA 105:9099–9104PubMedCentralCrossRefPubMedGoogle Scholar
  12. Grimaldi P, Orlando P, Di Siena S, Lolicato F, Petrosino S, Bisogno T, Geremia R, De Petrocellis L, Di Marzo V (2009) The endocannabinoid system and pivotal role of the CB2 receptor in mouse spermatogenesis. Proc Natl Acad Sci USA 106:11131–11136PubMedCentralCrossRefPubMedGoogle Scholar
  13. Horvath B, Mukhopadhyay P, Kechrid M, Patel V, Tanchian G, Wink DA, Gertsch J, Pacher P (2012) Beta-caryophyllene ameliorates cisplatin-induced nephrotoxicity in a cannabinoid 2 receptor-dependent manner. Free Radic Biol Med 52:1325–1333PubMedCentralCrossRefPubMedGoogle Scholar
  14. Legault J, Pichette A (2007) Potentiating effect of beta-caryophyllene on anticancer activity of alpha-humulene, isocaryophyllene and paclitaxel. J Pharm Pharmacol 59:1643–1647CrossRefPubMedGoogle Scholar
  15. Lucio RA, Tlachi-Lopez JL, Eguibar JR, Agmo A (2013) Sperm count and sperm motility decrease in old rats. Physiol Behav 110–111:73–79CrossRefPubMedGoogle Scholar
  16. Maccarrone M (2013) Endocannabinoids as markers of sperm quality: hot spots. Front Endocrinol (Lausanne) 4:169Google Scholar
  17. McClain RM, Downing JC, Edgcomb JE (1989) Effect of metronidazole on fertility and testicular function in male rats. Fundam Appl Toxicol 12:386–396CrossRefPubMedGoogle Scholar
  18. Molina-Jasso D, Alvarez-Gonzalez I, Madrigal-Bujaidar E (2009) Clastogenicity of beta-caryophyllene in mouse. Biol Pharm Bull 32:520–522CrossRefPubMedGoogle Scholar
  19. Mudry MD, Palermo AM, Merani MS, Carballo MA (2007) Metronidazole-induced alterations in murine spermatozoa morphology. Reprod Toxicol 23:246–252CrossRefPubMedGoogle Scholar
  20. Narayana K, Prashanthi N, Nayanatara A, Kumar HH, Abhilash K, Bairy KL (2005) Effects of methyl parathion (O,O-dimethyl O-4-nitrophenyl phosphorothioate) on rat sperm morphology and sperm count, but not fertility, are associated with decreased ascorbic acid level in the testis. Mutat Res 588:28–34CrossRefPubMedGoogle Scholar
  21. Narayana K, Verghese S, Jacob SS (2009) l-ascorbic acid partially protects two cycles of cisplatin chemotherapy-induced testis damage and oligo-astheno-teratospermia in a mouse model. Exp Toxicol Pathol 61:553–563CrossRefPubMedGoogle Scholar
  22. Page ST, Amory JK, Bremner WJ (2008) Advances in male contraception. Endocr Rev 29:465–493PubMedCentralCrossRefPubMedGoogle Scholar
  23. Rapino C, Battista N, Bari M, Maccarrone M (2014) Endocannabinoids as biomarkers of human reproduction. Hum Reprod Update 20:501–516CrossRefPubMedGoogle Scholar
  24. Rossi P, Lolicato F, Grimaldi P, Dolci S, Di Sauro A, Filipponi D, Geremia R (2008) Transcriptome analysis of differentiating spermatogonia stimulated with kit ligand. Gene Expr Patterns 8:58–70CrossRefPubMedGoogle Scholar
  25. Schuel H, Goldstein E, Mechoulam R, Zimmerman AM, Zimmerman S (1994) Anandamide (arachidonylethanolamide), a brain cannabinoid receptor agonist, reduces sperm fertilizing capacity in sea urchins by inhibiting the acrosome reaction. Proc Natl Acad Sci USA 91:7678–7682PubMedCentralCrossRefPubMedGoogle Scholar
  26. Seed J, Chapin RE, Clegg ED, Dostal LA, Foote RH, Hurtt ME, Klinefelter GR, Makris SL, Perreault SD, Schrader S, Seyler D, Sprando R, Treinen KA, Veeramachaneni DN, Wise LD (1996) Methods for assessing sperm motility, morphology, and counts in the rat, rabbit, and dog: a consensus report. Ilsi risk science institute expert working group on sperm evaluation. Reprod Toxicol 10:237–244CrossRefPubMedGoogle Scholar
  27. Tahtamouni LH, Alqurna NM, Al-Hudhud MY, Al-Hajj HA (2011) Dandelion (taraxacum officinale) decreases male rat fertility in vivo. J Ethnopharmacol 135:102–109CrossRefPubMedGoogle Scholar
  28. Whan LB, West MC, McClure N, Lewis SE (2006) Effects of delta-9-tetrahydrocannabinol, the primary psychoactive cannabinoid in marijuana, on human sperm function in vitro. Fertil Steril 85:653–660CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Zina M. Al-Alami
    • 1
  • Ziad A. Shraideh
    • 2
  • Mutasem O. Taha
    • 3
  1. 1.Faculty of PharmacyIsra UniversityAmmanJordan
  2. 2.Department of Biological Sciences, Faculty of ScienceThe University of JordanAmmanJordan
  3. 3.Department of Pharmaceutical Sciences, Faculty of PharmacyThe University of JordanAmmanJordan

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