Sains Malaysiana 48(4)(2019): 781–790

http://dx.doi.org/10.17576/jsm-2019-4804-10

 

Protective Effects of Zingiber zerumbet Ethyl Acetate Extract on Hydrogen Peroxide-Induced Damage of Red Blood Cells

(Kesan Perlindungan Ekstrak Etil Asetat Zingiber zerumbet pada Kerosakan Sel Darah Merah Aruhan Hidrogen Peroksida)

 

MOHD FUAD RAHMAT SAM^1,2, ASMAH HAMID¹, AHMAD ROHI GHAZALI¹, SANTHANA RAJ LOUIS³ & SITI BALKIS BUDIN¹*

 

¹Programme of Biomedical Science, Centre of Applied and Health Sciences, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Federal Territory, Malaysia

 

²Department of Biomedical Science, Kulliyyah of Allied Health Sciences, International Islamic University Malaysia, Jalan Istana, 25200 Kuantan, Pahang Darul Makmur, Malaysia.

 

³Unit of Electron Microscope, Institute for Medical Research, Jalan Pahang, 50588 Kuala Lumpur, Federal Territory, Malaysia

 

Received: 30 March 2018/Accepted: 28 January 2019

 

ABSTRACT

Zingiber zerumbet is widely used as therapeutic agent in traditional medicine and is reported to exert antioxidant activities. This study evaluated in vitro antioxidant potential of Z. zerumbet rhizome ethyl acetate extract on hydrogen peroxide (H2O2)-induced red blood cells (RBCs) damage; by measuring percentage of haemolysis, oxidative damage and morphological changes in treated RBCs. On a preliminary dose-response analysis, it was observed that incubation of RBCs with extract at doses of 50, 25, 12.5 and 6.25 μg/mL all had no significant effect on RBCs similar to NaCl control and BHT (positive control), therefore, these concentrations were used in the subsequent study. The RBCs were pre-incubated with extract in the chosen concentrations, prior to treatment with H2O2. The results showed that only 6.25 μg/mL+H2O2 group showed significantly (p<0.05) lower percentage of haemolysis and oxidative damage; indicated by low level of malondialdehyde (MDA) and protein carbonyls (PC) compared to the H2O2 alone group. Electron microscopic examination further showed that pre-treatment with 6.25 μg/mL extract reduced the H2O2 –induced morphological changes of RBCs. Phytochemical analysis of extract using Gas Chromatography Mass Spectrometry (GCMS) identified Zerumbone as the highest constituent compound (51.57%) in this extract. In conclusion, this study indicated that 6.25 μg/mL ethyl acetate extract Z. zerumbet rhizome could efficiently protect RBCs against oxidative damage induced by H2O2 and this effects could possibly due the high constituent of Zerumbone.

 

Keywords: Haemolysis; lipid peroxidation; red blood cells; Zingiber zerumbet

 

ABSTRAK

Zingiber zerumbet digunakan secara meluas sebagai agen terapeutik dalam perubatan tradisi dan dilaporkan mempunyai sifat antioksidan. Kajian ini dinilai melalui potensi in vitro antioksidan ekstrak Z. zerumbet rizom etil asetat pada hidrogen peroksida (H2O2)-teraruh kerosakan sel-sel darah merah (RBCs); dengan mengukur peratus hemolisis, kerosakan oksidatif dan perubahan morfologi untuk RBCs terawat. Analisis awalan dos gerak balas, dicerapi bahawa inkubator RBCs dengan ekstrak pada dos 50, 25, 12.5 dan 6.25 μg/mL tidak menunjukkan kesan ketara RBCs yang menyerupai kawalan NaCl dan BHT (kawalan positif), maka kepekatan ini digunakan dalam kajian yang seterusnya. RBCs telah pra-inkubasi dengan ekstrak dalam kepekatan pilihan, sebelum dirawat dengan H2O2. Hasil kajian menunjukkan bahawa hanya kumpulan 6.25 μg/mL + H2O2 menunjukkan signifikan (p<0.05) mengurangkan peratusan hemolisis dan kerosakan oksidatif; ditunjukkan oleh tahap rendah malondialdehid (MDA) dan protein karbonil (PC) berbanding kumpulan H2O2 sahaja. Pemeriksaan mikroskop elektron menunjukkan bahawa pra rawatan dengan ekstrak 6,25 μg/mL mengurangkan morfologi H2O2–teraruh perubahan RBCs. Analisis fitokimia menggunakan Kromatografi Gas Jisim Spektroskopi (GCMS) mengenal pasti zerumbon sebagai yang bentuk sebatian tertinggi (51.57%) di dalam ekstrak. Kesimpulannya, kajian ini menunjukkan bahawa 6.25 μg/mL ethil asetat rizom ekstrak Z. zerumbet secara cekap dapat melindungi RBCs daripada kerosakan oksidatif teraruh oleh H2O2 dan kesan ini boleh disebabkan oleh sebatian zerumbon yang tinggi.

 

Kata kunci: Hemolisis; pemperoksidaan lipid; sel-sel darah merah; Zingiber zerumbet

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*Corresponding author; email: balkis@ukm.edu.my