Antiparasitic effects of Elettaria cardamomum L. essential oil and its main compounds, 1-8 Cineole alone and in combination with albendazole against Echinococcus granulosus protoscoleces

Abstract

Background: The present investigation aims to determine the chemical structure and protoscolicidal effects of Elettaria cardamomum L. essential oil (ECEO) and its main compounds 1-8 cineole alone and along with albendazole (ALZ) against Echinococcus granulosus protoscoleces in vitro and ex vivo. We also decided to evaluate some cellular mechanisms such as the apoptotic activity and the permeability of plasma membrane of protoscoleces treated with ECEO and 1-8 cineole.

Methods: Hydatid cyst protoscoleces were divided into seven groups including protoscoleces treated with ECEO 50 µl/mL (T1), protoscoleces treated with ECEO 100 µl/mL (T2), protoscoleces treated with ECEO 200 µl/mL (T3), protoscoleces treated with 1-8 cineole 100 µg/mL (T4), protoscoleces treated with 1-8 cineole 200 µg/mL (T5), protoscoleces treated with 1-8 cineole 100 µg/mL + albendazole 50 µg/mL (T6), and protoscoleces treated with 1-8 cineole 200 µg/mL + albendazole ALZ-50 µg/mL (T7). The viability of protoscoleces were recorded by eosin staining examination. Moreover, the induction of apoptosis and the plasma membrane permeability of the protoscoleces treated with ECEO and 1-8 cineole were evaluated.

Results: The highest protoscolicidal effect of ECEO was observed at the dose of 200 µl/ml (T3). 1,8-Cineole alone and combined with ALZ, particularly at the dose of 200 µg/ml (T5 and T7), destroyed the 100% protoscolices after 10 min incubation. The ECEO (T1-T3) and 1-8 cineole alone (T4 and T5) and in combination with ALZ (T6 and T7) took longer to display their protoscolicidal effect ex vivo. The obtained results of relative fuorescent items exhibited that the protoscoleces incubated with ECEO and 1,8-Cineole, alter the permeability of plasma membrane by Sytox Green with increasing the concentration. The findings revealed exhibited that ECEO and 1,8-Cineole increasingly and dose-dependently induced activation of caspase-3 enzyme ranging from 6.8 to 23.3%.

Conclusion: Our obtained results revealed that ECEO and its main compound, 1,8-Cineole exhibited the potent protoscolicidal in vitro and ex vivo; and if more research is done on their efficacy and toxicity in animal models and even clinical setting, it can be suggested as a protoscolicidal agent to use during hydatid cyst surgery.

Keywords: Apoptosis; Cardamom; Cystic echinococcosis; Herbal medicines; Plasma membrane.

Fig. 1

Map of study site in Riyadh Province, Saudi Arabia.

Fig. 2

In vitro protoscolicidal activity of (A) various concentration of E. cardamomum essential oil (ECEO) and its main components, (B) 1,8-Cineole alone and in combined with albendazole (ALZ, 50 µg/ml) against hydatid cyst protoscoleces following various exposure times. Data are expressed as mean ± SD. (n = 3).

Fig. 3

Ex vivo protoscolicidal activity of (A) various concentration of E. cardamomum essential oil (ECEO) and its main components, (B) 1,8-Cineole alone and in combined with albendazole (ALZ, 50 µg/ml) against hydatid cyst protoscoleces following various exposure times. Data are expressed as mean ± SD. (n = 3).

Fig. 4

The plasma membrane permeability of the protoscoleces treated with various concentration of E. cardamomum essential oil (ECEO) and its main components, 1,8-Cineole. Data are expressed as mean ± SD. (n = 3).

Fig. 5

Evaluation of the induction of apoptosis in hydatid cyst protoscoleces by assess Caspase-3 activity of protoscoleces treated with various concentration of E. cardamomum essential oil and its main components, 1,8-Cineole. Data show as mean ± SD from three experiments in duplicate. * p < 0.05 difference was significant compared with control group.