A preventive effect of the combination of albendazole and pomegranate peel aqueous extract treatment in cystic echinococcosis mice model: An alternative approach

doi:10.1016/j.actatropica.2019.105050

Acta Tropica

Volume 197, September 2019, 105050

https://doi.org/10.1016/j.actatropica.2019.105050 Get rights and content

Abstract

To investigate the preventive effect of the combination of albendazole (ABZ) and pomegranate peel aqueous extract (PGE) treatment in cystic echinococcosis, we assess in vivo the antihydatic and the anti-inflammatory effects of the combination of ABZ/ PGE in cystic echinococcosis mice model. To evaluate the in vivo efficacy, mice were inoculated intraperitoneally with viable protoscolices and then treated with ABZ and/or PGE during cystic echinococcosis development. Mice were randomly allocated into eight groups: ABZ/CE group, PGE/CE group, (ABZ+PGE)/CE group, CE group, and control groups (Ctrl, PBS, ABZ, and PGE groups). Drugs in diverse treated groups were orally administered daily during CE development for two months. Mice were then euthanized and associated indications were investigated to evaluate the therapeutic efficacy. Cyst development and hepatic damage were macroscopically and histologically analyzed. The hepatic expression of iNOS, TNF-α, NF-κβ, vimentin, and CD68 was examined. Interestingly, the association of ABZ and PGE enhanced a significant reduction of the rate of hydatid cyst growth inhibition in comparison to the infected or ABZ-treated groups. This effect was strongly related to the histological structure of liver improvement. A significant iNOS, TNF-α, NF-κβ, vimentin, and CD68 decrease expression was observed in liver tissue of (ABZ+PGE)-treated group compared with infested and ABZ-treated groups. PGE treatment indicates a significant beneficial additive antihydatic effect with a reduction of the liver side effects. The combination of albendazole and PGE treatment is more efficient and suggests its potential preventive value against Echinococcus granulosus infection.

Introduction

Cystic echinococcosis (CE) is a chronic zoonosis affecting humans as well as domestic animals (Mandal et al., 2012). CE constitutes a serious public health problem in central Asia and China, South America, and Mediterranean countries (Deplazes et al., 2017). E. granulosus infection is characterized by a prolonged coexistence of the parasite and the host with no effective rejection reaction, although specific cellular and humoral immune responses are present (Margutti et al., 2002).

Currently, treatment options for CE include active surveillance, antiparasitic and surgery, including percutaneous treatment. The treatments depend on the type of the cysts. In fact, resolution of CE in non-surgical way with albendazole is confirmed to be effective in asymptomatic carriers with CE1 or CE3a cysts (Larrieu et al., 2018). WHO-IWGE (Meeting of the WHO Informal Working Group on Echinococcosis, 2017) mostly recommends ABZ for the treatment of cystic and alveolar echinococcosis. This is the only option in many inoperable cases like cyst in brain or multiple cysts and in the immune-depressed host. Nevertheless, this drug is not efficient in some cases and leads to liver toxicity and other side effects (Brunetti et al., 2010). Development of synergistic combinations of drugs, e.g. with albendazole (ABZ), can overcome toxicity and other side effects associated with high doses and/or long time dosage of single drugs (Siles-Lucas et al., 2018).

Alternative drugs and several natural compounds have been tested in in vitro and in vivo models of the Echinococcus species complex, but only few have reached clinical use (Hemphill et al., 2014). Punica granatum L. (common name pomegranate; subfamily Punicaceae) is a small tree originating from Asia and now widely cultivated in the Mediterranean basin, particularly in Algeria. Pomegranate peels or skin or rind are underestimated as an agricultural waste, though it is part of an ancient fruit with exceptionally rich ethnomedical applications and astringent properties (Middha et al., 2013). Many researchers have focused on the biological waste part of pomegranate for the purpose of discovering many beneficial effects for human health. The potential therapeutic properties of Pomegranate peels are wide-ranging and include treatment and prevention for cancer (Dikmen et al., 2011), cardiovascular disease (Jurenka, 2008), and diabetes (Middha et al., 2012), protection from ultraviolet radiation (Kanatt et al., 2010), and antimicrobial (Hayouni et al., 2011). Other potential applications include infant brain ischemia, Alzheimer's disease (Middha et al., 2012), male infertility, obesity, arthritis (Kanatt et al., 2010), anthelmintic and anti-protozoan activities (Calzada et al., 2006; El-Sherbini et al., 2009), and has shown efficacy in the treatment of parasitic worm infections (Pradhan et al., 1992). Previous works of our team reported the scolicidal, antihydatic and immunomodulatory effect of Pomegranate peel aqueous extract (PGE) (Labsi et al., 2016).

Several studies have demonstrated the therapeutic antioxidant and anti-inflammatory effects of P. granatum fruit, peel, and juice. These effects are mainly exerted by molecules like polyphenols and tannins (Gracious et al., 2001; Kaur et al., 2006; Moneim, 2012). In addition, it was ascertained that pomegranate peel possesses certain hepato-protective properties making it an important therapeutic agent in the treatment of fibrosis and oxidative damage (Toklu et al., 2007; Xu et al., 2008).

In this study and in the basis of our previous work, we investigate the combinatory effect of albendazole with Pomegranate in the murine model of echinococcosis. The antihydatic and immunomodulatory effects are analyzed in each case: ABZ treatment, PGE treatment and combinated ABZ/PGE treatment. In this regard, we evaluated the preventive effect of ABZ and/or PGE on the development of murine echinococcosis and on the integrity of liver function and architecture. Hepatic histological changes and inflammatory responses are tested such as the presence of glycoproteins and hepatic expression of iNOS, NF-κβ, TNF-α, CD68, and vimentin.

Section snippets

Collection of protoscoleces and initiation of experimental echinococcosis

E. granulosus PSC collection was realized as previously described (Amri and Touil-Boukoffa, 2015). PSC viability was assessed prior to inoculation and determined by 0.1% eosin staining. All samples exhibited viability >98% at the time of the experiment. Swiss mice were inoculated intraperitoneally (i.p.) with 2000 viable PSCs resuspended in sterile PBS (Urrea-Paris et al., 2002; Labsi et al., 2016; Khelifi et al., 2017; Labsi et al., 2018).

Experimental design

Female Swiss Albino mice (4–6 weeks old) were purchased

Effect of albendazole and/or Pomegranate peel aqueous extract on clinical parameters of experimental echinococcosis

Three months post-infection, 100% of animals from CE and PGE/CE groups developed cysts in the liver and other intraperitoneal areas. In ABZ/CE and ABZ+PGE/CE groups the infectivity was 77.8% and 44.4%, respectively. Hydatid cystic liver localization has dominated for all groups.

The diameter and the weight of developed cysts were measured to evaluate the effect of the combinatory therapy (ABZ, PGE) on cystic growth (Table 1). The mean of surface and weight of hydatid cysts were significantly

Discussion

The liver is the major site of CE involvement. The infection induces an immune imbalance on the hepatic tissue, leading to severe destruction of the architecture due to intensive inflammatory cell infiltrates and formation of fibrosis (Zhang et al., 2003). Inflammation and immunopathology is scarce, indicating that the parasite actively modulates the host innate and immune reaction. Similar to malignant tumors, metastasis formation into other organs can take place at a later stage of infection (

Conflict of interest statement

The authors declare that they have no competing interests.

Competing interests

The authors declare that they have no competing interests.

Acknowledgements

The author would like to thank Mohamed-Amine Lazourgui for his important contribution in histological study and for his technical help.

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A preventive effect of the combination of albendazole and pomegranate peel aqueous extract treatment in cystic echinococcosis mice model: An alternative approach

Abstract

Introduction

Section snippets

Collection of protoscoleces and initiation of experimental echinococcosis

Experimental design

Effect of albendazole and/or Pomegranate peel aqueous extract on clinical parameters of experimental echinococcosis

Discussion

Conflict of interest statement

Competing interests

Acknowledgements

Exp. Parasitol.

Acta Trop.

Acta Trop.

J. Ethnopharmacol.

Adv. Parasitol.

Comp. Immunol. Microbiol. Infect. Dis.

Phytomedicine.

Trends Pharmacol. Sci.

Food Chem. Toxicol.

Acta Trop.

Neuropharmacology.

Asian Pac. J. Trop. Med.

Acta Trop.

Nat. Prod. Res.

Asian Pac. J. Trop. Biomed.

Vet. Parasitol.

Res. Vet. Sci.

J. Hepatol.

Vet. Parasitol.

The antioxidant potency of Punica granatum L. Fruit peel reduces cell proliferation and induces apoptosis on breast cancer

J. Med. Food

Efﬁcacy of two plant extracts against vaginal trichomoniasis

J. Egypt. Soc. Parasitol.

Immunomodulatory activity of Punica granatum in rabbits-a preliminary study

J. Ethnopharmacol.

Treatment of echinococcosis: albendazole and mebendazole-what else?

Parasite.