Sains Malaysiana 50(4)(2021): 967-987

http://doi.org/10.17576/jsm-2021-5004-08

Phytochemical Profile, Antioxidant and Anti Proliferative Studies in Different Extracts of Artocarpus kemando Miq. Bark

(Kajian Profil Fitokimia, Antioksidan dan Anti Proliferatif dalam Ekstrak Berbeza Batang Artocarpus kemando Miq.)

NOOR SHAFIFIYAZ MOHD YAZID¹*, NAJIHAH MOHD HASHIM¹, HAPIPAH MOHD AL^I2 & RUSEA GO³

¹Department of Pharmacy, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Federal Territory, Malaysia

²Department of Chemistry, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Federal Territory, Malaysia

³Department of Biology, Faculty of Science, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia

Received: 20 September 2019/Accepted: 22 September 2020

ABSTRACT

In this study, the stem bark of Artocarpus kemando was used to find alternative antioxidants from natural sources with fewer side effects. A. kemando was extracted successively using hexane, chloroform and methanol solvents, and evaluated for antioxidant, cytotoxic, and antiproliferative activities. The extracts were investigated for determination of their total phenolic content (TPC) and total flavonoid content (TFC). Then, the antioxidant activities were evaluated using chemical based assays such as ferric reducing antioxidant power (FRAP), total antioxidant capacity, radical scavenging of 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) and 2,2'-azino-bis (3-ethylbenzothia zoline-6-sulphonic) (ABTS), β-carotene-linoleic acid (BC) assay, oxygen radical absorbance capacity (ORAC), and cell based assay. The cytotoxic study was done using four different cell lines namely human estrogen receptor positive (ER+) breast cancer cell line (MCF7), human ovarian cancer cell line (CAOV-3), human promyelocytic leukemia cell line (HL60), and normal immortalised human ovarian surface epithelial cell line (TI074), and were evaluated using microculture tetrazolium salt (MTT) before morphological change study was done on CAOV-3 cell. In this study, methanol extract displayed the most promising antioxidant activity compared to other extracts when tested with DPPH, FRAP, ABTS, TAOC, BC, ORAC, and cytoprotective assays. The remarkable activity showed by the methanol extract might be due to its high content of phenolic and flavonoid compounds at 855.5 ± 0.01 GAE µg/mL and 145.45 ± 0.06 QAE µg/mL, respectively. Nevertheless, the chloroform extract displayed better scavenging activity compared to other extracts with IC₅₀ value of 618 ± 0.04 µg/mL in DPPH assay. Each extract was analysed using Gas Chromatography Mass Spectrophotometry and the chemical constituents obtained were then analysed. In the cytoprotective activity, the methanol extract showed a comparable cytoprotection with ascorbic acid against the free radicals at the lowest effective concentration (EC₅₀) value of 21.48 µg/mL. However, in the cytotoxicity study, only chloroform extract displayed significant toxicity against the cancer cells with IC₅₀ value of 27.9 ± 0.03, 24.1 ± 0.02 and 9.0 ± 0.04 µg/mL after treatment at 24, 48, and 72 h, respectively. The chloroform extract of A. kemando was found capable of inducing apoptosis as shown with cell membrane blebbing, chromatin condensation and formation of apoptotic bodies. The results obtained from the study showed that A. kemando bark could be a potential antioxidant and antitumor agents particularly on human ovarian cancer cells.

Keywords: Antioxidant; antitumour; Artocapus kemando; cytoprotective; Malaysia

ABSTRAK

Dalam kajian ini, kulit batang Artocarpus kemando digunakan untuk mencari antioksidan alternatif daripada sumber semula jadi dengan kesan sampingan yang kurang. Kulit batang pokok A. kemando diekstrak secara berturutan menggunakan pelarut heksana, kloroform dan metanol serta dinilai untuk aktiviti antioksidan, sitotoksik dan antiproliferatif. Kesemua ekstrak telah dikaji untuk penentuan kandungan jumlah fenolik (TPC) dan jumlah flavonoid (TFC). Kemudian, aktiviti antioksidan telah dinilai melalui ujian berasaskan kimia seperti keupayaan antioksida mengurangkan ferum (FRAP), jumlah kapasiti antioksidan (TAOC), asai pengurangan radikal bebas menggunakan 2,2-difenil-1-pikrilhidrazil (DPPH) dan 2,2'-azino-bis (3-etilbenzotiazolin-6-sulfonik) (ABTS), asai asid β-karoten-linoleik (BC), kapasiti penyerapan oksigen radikal (ORAC), dan ujian berasaskan sel. Kajian sitotoksik dilakukan menggunakan empat titisan sel yang berbeza iaitu titisan sel kanser payudara positif reseptor estrogen (ER +) (MCF7), titisan sel kanser ovari manusia (CAOV-3), titisan sel leukemia promoelositik manusia (HL60) dan titisan sel normal permukaan epitelial ovari manusia (TI074), serta dinilai menggunakan garam mikro tetrazolium (MTT) sebelum kajian perubahan morfologi dilakukan terhadap sel CAOV-3. Dalam kajian ini, ekstrak metanol menunjukkan aktiviti antioksidan yang paling baik berbanding ekstrak lain apabila diuji dengan DPPH, FRAP, ABTS, TAOC, BC, ORAC dan asai sitoprotektif. Aktiviti luar biasa yang ditunjukkan oleh ekstrak metanol mungkin disebabkan oleh kandungan sebatian fenolik dan flavonoid yang tinggi iaitu masing-masing 855.5 ± 0.01 GAE μg/mL dan 145.45 ± 0.06 QAE μg/mL. Walau bagaimanapun, ekstrak kloroform menunjukkan aktiviti pengurangan radikal yang lebih baik berbanding ekstrak lain dengan nilai IC₅₀ 618 ± 0.04 μg/mL dalam asai DPPH. Setiap ekstrak telah dianalisis menggunakan Spektrofotometri Jisim Kromatografi Gas dan komponen kimia yang diperoleh kemudiannya dianalisis. Bagi aktiviti sitoprotektif, ekstrak metanol menunjukkan keupayaan sitoperlindung setanding dengan aktiviti asid askorbik terhadap radikal bebas pada nilai kepekatan berkesan yang paling rendah (EC₅₀) iaitu 21.48 μg/mL. Walau bagaimanapun, dalam kajian sitotoksisiti, hanya ekstrak kloroform yang menunjukkan ketoksikan signifikan terhadap sel kanser dengan nilai IC₅₀ masing-masing sebanyak 27.9 ± 0.03, 24.1 ± 0.02 dan 9.0 ± 0.04 μg/mL selepas rawatan pada 24, 48 dan 72 jam. Ekstrak chloroform A. kemando didapati berupaya mengaruh apoptosis sebagaimana ditunjukkan dengan penunasan membran sel, pengecutan kromatin dan pembentukan jasad apoptosis. Hasil yang diperoleh daripada kajian menunjukkan bahawa batang pokok A. kemando berpotensi sebagai agen antioksidan dan antitumor terutama ke atas sel kanser ovari manusia.

Kata kunci: Antioksidan; antitumor; Artocarpus kemando; Malaysia; sitoperlindung

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*Corresponding author; email: shafifiyaz63@gmail.com