Sains Malaysiana 47(9)(2018): 2129–2139

http://dx.doi.org/10.17576/jsm-2018-4709-22

Neuroprotective Effects of Ocimum basilicum Extract against Hydrogen Peroxide-Induced Oxidative Stress in SK-N-SH Neuroblastoma Cells

(Kesan Neuromelindungi Ekstrak Ocimum basilicum terhadap Tekanan Oksidatif yang Diaruh Hidrogen Peroksida dalam Sel Neuroblastoma SK-N-SH)

MANALI HANITI MOHD-ZAHID¹, JURIYATI JALIL¹, KOK MENG CHAN² & NORAZRINA AZMI^1*

¹Drug and Herbal Research Centre, Faculty of Pharmacy, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Federal Territory, Malaysia

²Toxicology Laboratory, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Federal Territory, Malaysia

Diserahkan: 4 Mac 2018/Diterima: 14 Mei 2018

ABSTRACT

Neurodegenerative diseases such as Alzheimer’s and Parkinson’s disease are characterized by the progressive loss of neurons. One of the contributing factors for these diseases is oxidative stress, characterized by the imbalance of free radicals production and antioxidant defense mechanisms. In the present study, the neuroprotective effects of Ocimum basilicum var. thyrsiflora against hydrogen peroxide (H₂O₂)-induced oxidative stress in SK-N-SH neuroblastoma cells were evaluated. The exposure of SK-N-SH cells to 50 μM H2O2 for 24 h induced cytotoxicity and apoptosis as measured by cell viability and flow cytometry, respectively. Pretreatment with ethyl acetate (ObEA) fraction at 3.1-25 μg/mL showed the highest protection against H₂O₂-induced cell death compared to other fractions and crude extract by increasing cell viability and reducing apoptosis. The evaluation of antioxidant capacity via 1, 1-diphenyl-2-picrylahydrazyl (DPPH) and ferric reducing/antioxidant power (FRAP) assays showed ObEA possessed the highest antioxidative properties. The intracellular reactive oxygen species (ROS) production of H₂O₂ in untreated cells increased by 2.39-fold compared to the control and was significantly attenuated by the 2 h pre-treatment of O. basilicum (p<0.05). The reduction in intracellular superoxide dismutase (SOD) induced by H₂O₂ was also abrogated by the pretreatment of O. basilicum. These findings suggested that O. basilicum is potentially neuroprotective against oxidative damage in neuronal cells by scavenging free radicals, restoring SOD activities and eventually prevent cell death.

Keywords: Antioxidant; neuroprotective; Ocimum basilicum; rosmarinic acid

ABSTRAK

Penyakit kemerosotan saraf seperti Alzheimer dan Parkinson dicirikan oleh kehilangan neuron secara progresif. Salah satu faktor penyumbang penyakit ini ialah tekanan oksidatif, yang dicirikan oleh ketidakseimbangan antara penghasilan radikal bebas dan mekanisme pertahanan antioksidan. Dalam kajian ini, kesan neuromelindungi Ocimum basilicum var. thyrsiflora terhadap tekanan oksidatif yang diaruh hidrogen peroksida (H₂O₂) dalam sel neuroblastoma SK-N-SH telah dinilai. Pendedahan sel SK-N-SH terhadap 50 μM H₂O₂ selama 24 jam telah menyebabkan sitotoksik dan apoptosis yang masing-masing diukur melalui kebolehhidupan sel dan aliran sitometri. Pra-rawatan dengan fraksi etil asetat (ObEA) pada 3.1-25 μg/mL menunjukkan kesan perlindungan tertinggi terhadap kematian sel yang diaruh H₂O₂ berbanding fraksi lain dan ekstrak mentah dengan meningkatkan kebolehhidupan sel dan mengurangkan apoptosis. Penilaian kapasiti antioksidan melalui asai difenil-pikrilhidrazil (DPPH) dan asai kuasa antioksidan penurunan ferik (FRAP) menunjukkan ObEA mempunyai ciri antioksidan tertinggi. Pengeluaran spesies oksigen reaktif (ROS) intrasel dalam sel yang hanya dirawat H₂O₂ meningkat sebanyak 3.29 kali ganda berbanding dengan sel kawalan dan dilemahkan secara signifikan oleh 2 jam pra-rawatan O. basilicum (p<0.05). Pengurangan superoksida dismutase (SOD) intrasel yang disebabkan oleh H₂O₂ juga turut dilemahkan oleh pra-rawatan O. basilicum. Keputusan kajian ini mencadangkan bahawa O. basilicum berpotensi berfungsi sebagai agen neuromelindungi terhadap kerosakan oksidatif dalam sel-sel neuron dengan merencatkan radikal bebas, memulihkan aktiviti SOD dan akhirnya menghalang kematian sel neuron.

Kata kunci: Antioksidan; asid rosmarinik; neuromelindungi; Ocimum basilicum

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*Pengarang untuk surat-menyurat; email: azrina.azmi@ukm.edu.my