Sains Malaysiana 40(6)(2011): 595–599

Allicin-Induced Cellular Destruction of Haemoflagellate Protozoa, Trypanosoma evansi In Mice

(Kemusnahan Sel Protozoa Hemoflagelat Trypanosoma evansi Aruhan Alisin dalam Mencit)

B. A. H. Zainal-Abidin* & Mohd Shukri Hj Baba

Pusat Pengajian Biosains & Bioteknologi, Fakulti Sains & Teknologi

Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor D.E., Malaysia

Received: 21 May 2010 / Accepted: 25 October 2010

ABSTRACT

Allicin or diallyl thiosulfinate (C3H5SS(O)C3H5), an active compound of garlic (Allium sativum) is known for its pharmaceutical properties. In this study, the cellular destructive effects of allicin on a haemoflagellate protozoa parasite Trypanosoma evansi was investigated. Groups of male ICR mice were infected with a lethal dose of the parasite (1×105 parasites per mouse) and each group was either treated intraperitoneally with berenil (0.01 mL per mouse, a commercial anti-trypanosomal drug) on D+3 post-infection as the positive control group, treated orally with allicin (0.1 mL of 15 μg/mL allicin solution per mouse) for 30 days starting from D-7 pre-infection as the experimental group, or left untreated as the negative control group. Thin-stained blood smears were prepared from each mouse every alternate day, starting from D+3 post-infection and continued until the animal succumbed or until D+90 post-infection. Parasitaemias were determined using light microscope. Unstained blood smears were also prepared for direct observation under Phillips XL30 and Leo 1450VP scanning electron microscopes. All mice in the negative group succumbed to the infection with drastic increase of parasitaemias while all the positive control mice had minimal parasitaemias and cleared from the infection and survived for more than 100 days. On the other hand mice in the experimental group, experienced a prolonged suppressed parasitaemias which became patent later and caused death to all mice. Micrograph observations of parasites in the positive group showed that the parasites had adverse morphological changes due to berenil treatment which lead to cell destruction and death within 5 – 6 hours post-treatment. Likewise parasites in the experimental group too had undergone profound physical damages which caused cell death. This is the first report which shows that allicin actually induced cellular damage to haemoflagellate cells of T. evansi in vivo.

Keywords: Allicin; cellular destruction; haemoflagellate protozoa; parasitaemia; Trypanosoma evansi

ABSTRAK

Alisin atau ‘diallyl thiosulfinate’ (C3H5SS(O)C3H5) adalah sebatian aktif bawang putih (Allium sativum) yang mempunyai banyak khasiat farmaseutikal. Dalam kajian ini kesan kemusnahan alisin ke atas sel parasit protozoa hemoflagelat Trypanosoma evansi telah diselidiki. Kumpulan-kumpulan mencit jantan strain ICR telah dijangkitkan dengan takaran maut parasit (1×105 parasit per mencit) dan setiap kumpulan mencit sama ada dirawat dengan berenil iaitu sejenis dadah anti-tripanosom kemersial secara suntikan intraperitonem pada hari D+3 pasca-jangkitan (kumpulan kawalan positif), atau mencit dirawat dengan alisin secara oral (0.1 mL of 15 μg/mL larutan alisin per mencit) selama 30 hari bermula pada hari D-7 pasca-jangkitan (kumpulan kajian), atau mencit tidak mendapat sebarang rawatan (kumpulan kawalan negatif). Slaid darah calitan nipis terwarna disediakan daripada setiap ekor mencit berselang hari bermula pada hari D+3 sehingga mencit maut atau sehingga hari D+90 pos-jangkitan. Parasitemia telah ditentukan di bawah mikroskop cahaya. Calitan darah tak berwarna turut disediakan untuk cerapan langsung menggunakan mikroskop elektron pengimbasan Phillips XL30 dan Leo 1450VP. Didapati semua mencit kumpulan kawalan negatif mati akibat peningktan parasitemia yang cepat manakala mencit dalam kumpulan kawalan positif menunjukkan parasitemia yang minima dan bebas daripada jangkitan parasit dan bermandiri melebihi 100 hari. Sebaliknya mencit dalam kumpulan kajian mengalami perkembangan parasitemia tertindas yang kemudiannya meningkat semula dan menyebabkan kematian mencit. Cerapan ke atas mikrograf darah daripada kumpulan kawalan positif menunjukkan morfologi parasit mengalami kerosakan teruk kesan tindakan berenil yang menyebabkan kemusnahan dan kematian sel parasit 5-6 jam pasca-rawatan. Sebagai perbandingan parasit daripada kumpulan kajian juga mengalami kerosakan fizikal yang parah dan membawa kepada kematian. Hasil kajian ini merupakan laporan pertama yang menunjukkan alisin mengaruh kemusnahan sel-sel parasit T. evansi secara in vivo.

Kata kunci: Alisin; kemusnahan sel; parasitemia; protozoa hemoflagelat; Trypanosoma evansi

REFERENCES

Ali, M., Thomson, M. & Afzal, M. 2000. Garlic and onions: their effect on eicosanoid metabolism and its clinical relevance. Prostaglandins Leukotrienes and Essential Fatty Acids 62: 55-73.

An, MM., Shen, H., Cao, YB., Zhang, JD., Cai, Y., Wang, R. & Jiang, Y.Y. 2009. Allicin enhances the oxidative damage effect of amphotericin B against Candida albicans. International Journal of Antimicrobial Agents 33: 258-263.

Anonymous. 2003. Universiti Kebangsaan Malaysia Animal Ethics Committee Guidelines (UKMAEC). UKMAEC Secretariat, Kuala Lumpur, 1-40.

Ariyanayagam, M.R. & Fairlamb, A.H. 2001. Ovothiol and trypanothione as antioxcidants in trypanosomatids. Molecular and Biochemical Parasitology 115: 189-198.

Mirelman, D., Monheit, D. & Varon, S. 1987. Inhibition of growth of Entamoeba histolytica by allicin, the active principle of garlic extract (Allium sativum). Journal of Infectious Diseases 156(1): 243-244.

Oommen, S., Anto, R.J., Srinivas, G. & Karunagaran, D. 2004. Allicin (from garlic) induces caspace-mediated apoptosis in cancer cells. European Journal of Pharmacology 485: 97-103.

Pilch, D.S. Kirolos, M.A. & Breslauer D.S. 1995. Berenil binding to higher ordered nucleic acid structures: complexion with a DNA and RNA triple helix. Biochemistry 34: 16107-16124.

Rabinkov, A., Miron, T., Konstaninovski, L., Wilchek, M., Mirelman, D. & Weiner, L. 1998. The mode of action of allicin: trapping of radicals and interaction with thiol containing proteins. Biochemicaet Biophysica Acta 1379: 233-244.

Ramos, F.A., Takaishi, Y., Shirotori, M., Kawaguchi, Y., Tsuchiya, K. & Shibata H. 2006. Antibacterial and antioxcidant activities of quercetin oxidation products from yellow anion (Allium cepa) skin. Journal of Agricultural and Food Chemistry 54: 3551-3557.

Romão, P.R.T., Tovar, J. Fonseca, S.G. Moraes, R.H., Cruz, A.K., Hothersall, J.S., -Noronha-Dutra, A.A., Ferreira, S.H. & Cunha, F.Q. 2006. Glutathione and the redox control system trypanothione/trypanothione reductase are involved in the protection of Leishmania spp. against nitrosothiol-induced cytotoxicity. Brazilian Journal of Medical and Biological Research 39(3): 355-363.

Saleehen, D., Ali, S.A. & Yasinzai M.M. 2004. Antileishmanial activity of aqueous onion extract in vitro. Fitoterapia 7(51): 9-13.

Verma, B.B., Gautam, O.P. & Malik P.D. 1976. Trypanosoma evansi: therapeutic efficacy of diaminazine aceturate in crossbred calves, Bos Taurus and B. indicus. Experimental Parasitology 40: 406-410.

Yamada, Y. & Azuma, K. 1997. Evaluation of the in vitro antifungal activity of allicin. Antimicrob. Agents Chemother. 11: 743-749.

Zainal-Abidin, B.A.H.1992. Infections of Trypanosoma evansi in Malaysia. Malaysian Applied Biology 2(10): 1-8.

*Corresponding author; e-mail: zainala@ukm.my