Synergistic Effects of Combined Caryophyllene and Albendazole against Invaginated and Evaginated Protoscoleces of Echinococcus granulosus
-
Maryam khodadadi
,
Mohammad Javad Pour Maram
,
Hossein Amini Khoei
,
Hadi Raeisi Shahraki
,
Ebrahim Saedi Dezaki
Abstract
Background: Natural compounds have garnered increasing attention as candidate antiparasitic therapeutics. We aimed to evaluate the protoscolicidal efficacy of caryophyllene (CAP), alone and in combination with albendazole (ALZ), against hydatid cyst protoscoleces.
Methods: Suspensions of 10,000 invaginated and evaginated protoscoleces (400 µL normal saline) were exposed to CAP (62.5–500 µg/mL), ALZ (250 µg/mL), or CAP+ALZ combinations in 24-well culture plates. Protoscolex viability was assessed by eosin exclusion test. Reactive oxygen species (ROS) induction was quantified using the DCFH-DA assay, and pre-apoptotic gene expression was analyzed by real-time PCR.
Results: CAP, particularly in combination with ALZ, significantly reduced the viability of both invaginated and evaginated protoscoleces in a concentration- and time-dependent manner. Combination therapy achieved 100% mortality of invaginated protoscoleces within 10–30 minutes, demonstrating significant synergism over either monotherapy. Evaginated protoscolex viability was also markedly diminished. The observed scolicidal activity was mechanistically correlated with apoptosis induction, evidenced by elevated ROS production and significant upregulation of caspase-3 and P53 gene expression (P<0.001).
Conclusion: CAP, particularly in combination with ALZ, exhibits potent in vitro protoscolicidal activity against hydatid cyst protoscoleces, with apoptosis induction and ROS generation implicated as underlying mechanisms. Further pharmacodynamic investigations and clinical trials are warranted to elucidate the precise mechanisms of action and evaluate the therapeutic potential of CAP-based combination therapy in the management of cystic echinococcosis.
2. Khalili N, Iranpour P, Khalili N, Haseli S. Hydatid disease: a pictorial review of uncommon locations. Iran J Med Sci. 2023;48(2):118-129.
3. Dietrich CF, Douira-Khomsi W, Gharbi H, et al. Cystic echinococcosis, review and illustration of non-hepatic manifestations. Med Ultrason. 2020;22(3):319-24.
4. Ün M, Yaman SS, Erbaş O. Hydatid cyst and treatment. Demiroglu Science University Florence Nightingale Journal of Transplantation. 2020;5(2):070-8.
5. Cheraghipour K, Rouzbahani AK, Fallahi S, et al. Recent advances in therapeutic strategies against hydatid cysts using nanomaterials: A systematic review. Letters in Drug Design & Discovery. 2023 1;20(9):1185-93.
6. Abdelmohsen UR, Balasubramanian S, Oelschlaeger TA, et al. Potential of marine natural products against drug-resistant fungal, viral, and parasitic infections. Lancet Infect Dis. 2017;17(2):e30-41.
7. Gyrdymova YV, Rubtsova SA. Caryophyllene and caryophyllene oxide: A variety of chemical transformations and biological activities. Chemical Papers. 2022;76(1):1-39.
8. Blake K. Beta-Caryophyllene: A review of current research. Alternative and Complementary Therapies. 2021;27(5):222-6.
9. Francomano F, Caruso A, Barbarossa A, et al. β-Caryophyllene: a sesquiterpene with countless biological properties. Applied Sci. 2019;9(24):5420.
10. Sharma C, M. Al Kaabi J, M. Nurulain S, N. Goyal S, Amjad Kamal M, Ojha S. Polypharmacological properties and therapeutic potential of β-caryophyllene: a dietary phytocannabinoid of pharmaceutical promise. Curr Pharm Des. 2016;22(21):3237-64.
11. Mahmoudvand H, Kheirandish F, Dezaki ES, Shamsaddini S, Harandi MF. Chemical composition, efficacy and safety of Pistacia vera (var. Fandoghi) to inactivate protoscoleces during hydatid cyst surgery. Biomed Pharmacother. 2016;82:393-8.
12. Mahmoudvand H, Kheirandish F, Ghasemi Kia M, Tavakoli Kareshk A, Yarahmadi M. Chemical composition, protoscolicidal effects and acute toxicity of Pistacia atlantica Desf. fruit extract. Nat Prod Res. 2016;30(10):1208-11.
13. Mahmoudvand H, Asadi A, Harandi MF, Sharififar F, Jahanbakhsh S, Dezaki ES. In vitro lethal effects of various extracts of Nigella sativa seed on hydatid cyst protoscoleces. Iran J Basic Med Sci. 2014;17(12):1001–1006.
14. Marhoon IA, Alwan MS, Al-Ibrahimi LA. Promising Anthelminthic Effects of Camphene Against Protoscoleces and Hydatid Cysts of Echinococcus granulosus. Acta Parasitol. 2025;70(3):113.
15. Lu S, Wen L, Gong Y, Tian C, et al. In vitro effects of harmine against Echinococcus granulosus protoscoleces by stimulating DNA damage. Exp Parasitol. 2021;226-227:108121.
16. Mahmoudvand H, Khalaf AK, Moghadam EF, Yadegari JG, Adineh A, Jalili A. Investigating the Lethal Effects of Carvone on the Protoscoleces of Hydatid Cyst, In vitro and Ex Vivo. Iran J Parasitol. 2025;20(1):91-9.
17. Shang X, Dai L, Cao X, Ma Y, Gulnaz I, Miao X, Li X, Yang X. Natural products in antiparasitic drug discovery: advances, opportunities and challenges. Natural Product Reports. 2025; 42(9):1419-58.
18. Boța M, Vlaia L, Jîjie AR, et al. Exploring synergistic interactions between natural compounds and conventional chemotherapeutic drugs in preclinical models of lung cancer. Pharmaceuticals. 2024;17(5):598.
19. Maroufi Y, Hoseini SR, Alavi M. Antiparasitic effect of leaf extract and major metabolites of Pelargonium quercetorum Agnew. against Leishmania major: in vitro and in silico studies. Journal of Applied Biotechnology Reports. 2022;9(4):817-30.
20. Estevam EB, Deus IP, Silva VP, et al. In vitro antiparasitic activity and chemical composition of the essential oil from Protium ovatum leaves (Burceraceae). Anais da Academia Brasileira de Ciências. 2017;89(04):3005-13.
21. Ahmed EA. The Potential Therapeutic Role of Beta-Caryophyllene as a Chemosensitizer and an Inhibitor of Angiogenesis in Cancer. Molecules. 2025;30(8):1751.
22. Bonincontro G, Scuderi SA, Marino A, Simonetti G. Synergistic Effect of Plant Compounds in Combination with Conventional Antimicrobials against Biofilm of Staphylococcus aureus, Pseudomonas aeruginosa, and Candida spp. Pharmaceuticals. 2023;16(11):1531.
23. Lim CR, Ogawa S, Kumari Y. β-Caryophyllene: a comprehensive review of its pharmacological activities, mechanisms of action, and therapeutic potential. Archives of Pharmacal Research. 2025;48(1):1–42. doi: 10.1007/s12272-024-01523-z.
24. Moo CL, Yang SK, Osman MA, et al. Antibacterial activity and mode of action of β-caryophyllene on Bacillus cereus. Pol J Microbiol. 2020;69(1):1-6.
25. Cancela M, Paes JA, Moura H, Barr JR, Zaha A, Ferreira HB. Unraveling oxidative stress response in the cestode parasite Echinococcus granulosus. Sci Rep. 2019;9(1):15876.
26. Fidyt K, Fiedorowicz A, Strządała L, Szumny A. β‐caryophyllene and β‐caryophyllene oxide—natural compounds of anticancer and analgesic properties. Cancer Med. 2016;5(10):3007-17.
27. Buret AG, Bhargava A. Modulatory mechanisms of enterocyte apoptosis by viral, bacterial and parasitic pathogens. Crit Rev Microbiol. 2014;40(1):1-17.
28. Barcinski MA, DosReis GA. Apoptosis in parasites and parasite-induced apoptosis in the host immune system: a new approach to parasitic diseases. Braz J Med Biol Res. 1999;32:395-401.
29. Pawłowska M, Mila-Kierzenkowska C, Szczegielniak J, Woźniak A. Oxidative stress in parasitic diseases—Reactive oxygen species as mediators of interactions between the host and the parasites. Antioxidants. 2023;13(1):38.
30. Ullah H, Di Minno A, Santarcangelo C, Khan H, Daglia M. Improvement of oxidative stress and mitochondrial dysfunction by β-caryophyllene: A focus on the nervous system. Antioxidants. 2021;10(4):546.
| Files | ||
| Issue | Vol 21 No 2 (2026) | |
| Section | Original Article(s) | |
| Keywords | ||
| Natural products Scolicidal Apoptosis Protoscoleces Echinococcus granulosus | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
