Elsevier

Acta Tropica

Volume 98, Issue 2, May 2006, Pages 152-161
Acta Tropica

Cytotoxic effect of curcumin on Giardia lamblia trophozoites

https://doi.org/10.1016/j.actatropica.2006.03.005Get rights and content

Abstract

Giardia lamblia is one of the most important worldwide causes of intestinal infections produced by protozoa. Thus, the search for new alternative therapeutic approaches for this parasitic disease is very important. Common drugs used to control and eradicate this infection, frequently exhibit side effects that force patients to abandon treatment. The present work evaluates the anti-protozoan activity of curcumin, the main constituent of turmeric. Axenic G. lamblia (Portland 1 strain) cultures were exposed to different concentrations of curcumin. Its effects were evaluated on parasite growth, adhesion capacity and parasite morphology. We also evaluated the capacity of curcumin to induce an apoptosis-like effect. All curcumin concentrations inhibited trophozoite growth and adhesion in more than 50% in dose and time dependent manner. Morphological changes were described as protrusions formed under the cytoplasmic membrane, deformation due to swelling and cell agglutination. Curcumin induced apoptosis-like nuclear staining in dose and time dependent manner. In conclusion, curcumin exhibited a cytotoxic effect in G. lamblia inhibiting the parasite growth and adherent capacity, induced morphological alterations, provoked apoptosis-like changes. Future in vitro and in vivo experiments are endowed to elucidate the effect of curcumin in an experimental model of G. lamblia infection, analyze the involvement of ion channels in the swelling effect of curcumin during an apparent osmotic deregulation in G. lamblia trophozoites. This will lead to the proposal of the action mechanism of curcumin as well as the description of mechanism involved during the activation process for the apoptotic-like effect.

Introduction

Human and animal diseases caused by protozoan parasites have great impact on public health (Upcroft and Upcroft, 2001). Giardia lamblia, is one of the most common parasites found in the human intestinal tract which may cause an acute or chronic infection worldwide known as giardiosis. Clinical symptoms include: nausea, stomach cramps, diarrhea, vomiting, malabsorption syndrome, steatorrhea, weight loss, headache, low scholar progress, deficiencies in mental and physical development being more intense in childhood (Ridley and Ridley, 1976, Gardner and Hill, 2001, Ortiz et al., 2001).

A number of antigiardiosic drugs have been developed to control this infectious disease. Nitroimidazoles, as metronidazole, is known to induce DNA fragmentation and this may affect host cells, thus, its use during pregnancy is not recommended (Oxberry et al., 1994, Campanati and Monteiro-Leal, 2002). Benzimidazoles, as albendazole are drugs that affect parasite cytoskeleton causing parasite detachment from intestinal epithelium (Harder et al., 2001, Samuelson, 1999). Nitrofuran compounds, as furazolidone affect parasite adhesion capacity but not its viability, this drug is approved by FDA (Hoyne et al., 1989). Nitazoxamide is tiazolidic derivative that exhibits similar effects as metronidazole and has been tested for treatment of giardiosis (Ponce-Macotela et al., 2001, Ortiz et al., 2001). All anti-giardiosic drugs above mentioned cause side effects that frequently induce treatment interruption.

Curcumin is a yellow–orange dye derived from the rhizome of the plant Curcuma longa. Curcumin (diferuloyl methane, Fig. 1), one of the eleven curcuminoids identified by analytical methods, is the most abundant compound and exerts a variety of well documented effects. It is an anti-tumor agent with anti-inflammatory and anti-oxidant properties by inhibition of nuclear factor κ B and AP1 transcription which consequently decreases the expression of pro-inflammatory cytokines, inducible nitric oxide synthase, cyclooxygenase and lipoxygenase, which contribute to reduce the inflammation and radical oxidant synthesis (Huang et al., 1997, Chan et al., 1998, Skrzypezak-Jankun et al., 2000, Bhaumik et al., 2000, Lim et al., 2001, Kang et al., 1999, Pan et al., 2000).

Curcumin confers anti-oxidant protection against ethanol-induced lipid peroxidation in neuronal cells functioning as a scavenger for oxidant radicals (Rajakrishnan et al., 1999).

As anti-cancer, curcumin induces apoptosis in tumoral cell lines, in vivo chemically induced cancer and in clinical trails (Sharma et al., 2001, Shukla et al., 2003, Lin et al., 2001, Dorai et al., 2001). Curcumin also inhibits TPA-induced protein kinase C activity (Limtrakul et al., 1997).

Anti-genotoxic effects have also been documented against mutagenic and carcinogenic compounds (Limtrakul et al., 2001, Huang et al., 1997, Ishizaki et al., 1996).

The anti-atherogenic effect of curcumin has been demonstrated in an animal model of experimental atherosclerosis where the most striking fact is the anti-oxidant activity avoiding the oxidation of LDL-cholesterol as oxidized LDL-cholestrol is very injurious to blood vessels (Ramírez-Tortosa et al., 1999) Curcumin, also posses anti-bacterial (beta-lactamase positive) and anti-fungal activities (Apisariyakul et al., 1995, Wuthi-udomlert et al., 2003).

Anti-protozoal activities have been documented in in vitro studies against Leishmania major promastigotes (Rasmusen et al., 2000) and against leishmanial strains and also against amastigote like cells of L. major (Saleheen et al., 2002).

In this paper we describe the cytotoxic effect of curcumin against G. lamblia trophozoites, measuring parasite growth, adhesion capacity, viability, morphology and apoptotic-like changes.

Section snippets

Chemicals

Curcumin was purchased from Sigma Chemical Co., St. Louis, MO, purity: 65–70% and is an extract obtained from Curcuma longa.

Effect of curcumin on G. lamblia trophozoite growth

After seeding 1.5 × 105 trophozoites/ml, amount determined with a growth curve assay, the maximum amount of parasites was achieved after 72 h of incubation. Thus, the optimal times for analysis are 24, 48 and 72 h after seeding and initial exposure to curcumin.

G. lamblia trophozoites growth significantly decreased at all curcumin concentrations and at all times analyzed compared to the vehicle control group. Briefly, 0.3 μM of curcumin decreased G. lamblia growth at 24 h 31.7%, at 48 h 52% and at 72 h

Discussion

Turmeric is a spice traditionally used in Asian gastronomy. Curcumin is the most abundant curcuminoid extracted from this spice. This compound has been exhaustively studied since the early 1980's. The most striking biological effects of curcumin include anti-oxidant, anti-inflammatory, anti-genotoxic, anti-tumoral and anti-bacterial effects (Araújo and Leon, 2001). Recently, an anti-parasitic in vitro activity against Leishmania has been described (Saleheen et al., 2002, Koide et al., 2002).

Acknowledgements

This research work was supported by Universidad de Guadalajara, PROPESTI/PITI/65 2004, and represents the Doctoral Thesis of Pérez-Arriaga L.

Ramírez-Herrera M.A., Pérez-Arriaga L. and Cortés Zárate R. contributed equally for this research work.

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