Sains Malaysiana 48(8)(2019): 1737–1744
http://dx.doi.org/10.17576/jsm-2019-4808-20
Inhibitory Effects of
Gynura procumbens Ethanolic Extract on Nitric Oxide Production and Inducible
Nitric Oxide Synthase (iNOS) Protein
Expression in Macrophages (Kesan
Perencatan Ekstrak Etanol Gynura procumbens terhadap Penghasilan Nitrik Oksida dan
Ekspresi Protein Sintase
Nitrik Oksida Teraruh
(iNOS) di dalam
Makrofaj)
TAN JIAH NING1, SYARATUL DALINA YUSOFF1, ZAKIAH JUBRI2, FHATAHEYA BUANG1, TAN ZE SONG1, AMEERAH BUDIONO1, IBRAHIM JANTAN3, ROZA DIANITA4, ENDANG KUMOLOSASI1, NORAZRINA AZMI1 & NORSYAHIDA MOHD FAUZI1*
1Drug
and Herbal Research Centre, Faculty of Pharmacy, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia
2Department
of Biochemistry, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latiff, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia
3School
of Pharmacy, Taylor's University, Lakeside Campus, 47500 Subang
Jaya, Selangor Darul Ehsan, Malaysia
4School
of Pharmaceutical Sciences, Universiti Sains Malaysia, 11800 USM Pulau Pinang, Malaysia
Received: 14 September 2018/Accepted: 31 May
2019
ABSTRACT
Nitric oxide (NO)
overproduction by inducible nitric oxide synthase (iNOS)
may be associated with acute and chronic inflammations. Macrophages as
important cells in the innate immune system are able to be stimulated and can
lead to iNOS activation and excessive NO production. Gynura procumbens is a medicinal plant
traditionally used in treating various ailments including inflammation but the
mechanism of anti-inflammatory activity of this plant is still elusive. This
study was carried out to investigate the anti-inflammatory therapeutic effects
of Gynura procumbens ethanolic extract on NO production
and iNOS protein expression in RAW 264.7
macrophages stimulated with lipopolysaccharide (LPS).
Cell viability of RAW 264.7 macrophages treated with Gynura procumbens ethanolic extract was determined by MTT assay. NO production was determined by Griess assay following Gynura procumbens ethanolic extract treatment alone or in
combination with LPS stimulation. Protein expression of iNOS was determined by western blot. RAW 264.7
macrophages viability of more than 90% was observed after 24 h treatment with Gynura procumbens ethanolic extract concentration range of 3.9 μg/mL to 500 μg/mL. Significant inhibition of NO production
level has been identified in LPS-stimulated RAW 264.7
cells pre-treated with 250 μg/mL Gynura procumbens ethanolic extract (p<0.05) while all
selected concentrations of Gynura procumbens ethanolic extract showed no significant alteration of NO production
in the absence of LPS stimulation. Pre-treatment of 250 μg/mL Gynura procumbens ethanolic extract also demonstrated significant suppression of iNOS protein expression in LPS-stimulated RAW 264.7
cells (p<0.05). In conclusion, this study demonstrates that Gynura procumbens ethanolic extract exhibits anti-inflammatory
potential through inhibition of NO production and iNOS protein expression in LPS-stimulated
macrophages, suggesting that this plant could be further researched for its
beneficial use in inflammatory disorders.
Keywords: Gynura procumbens; inducible nitric oxide synthase; inflammation; lipopolysaccharide;
macrophage; nitric oxide
ABSTRAK
Penghasilan nitrik oksida
(NO)
yang berlebihan oleh
sintase nitrik oksida teraruh (iNOS) mungkin boleh
dikaitkan dengan
radang akut dan kronik. Makrofaj
sebagai sel
yang penting dalam sistem
keimunan inat
berupaya dirangsang dan menyebabkan aktivasi iNOS dan
penghasilan NO yang berlebihan.
Gynura procumbens ialah tumbuhan
perubatan tempatan yang digunakan secara tradisi untuk merawat pelbagai
jenis penyakit
termasuk radang namun mekanisme aktiviti anti-radang oleh tumbuhan ini
masih sukar
difahami. Kajian ini dijalankan untuk mengkaji kesan terapeutik anti-radang oleh ekstrak etanol Gynura procumbens terhadap penghasilan
NO
dan ekspresi protein iNOS dalam makrofaj
RAW
264.7 yang dirangsang
oleh lipopolisakarida (LPS).
Kebolehhidupan sel
makrofaj RAW 264.7 oleh
rawatan ekstrak
etanol Gynura procumbens ditentukan
dengan asai MTT.
Reagen Griess digunakan
untuk menentukan
penghasilan NO diikuti
rawatan ekstrak
etanol Gynura procumbens atau dengan gabungan rangsangan LPS. Ekspresi
protein iNOS dikaji
dengan pemblotan western. Di
bawah rawatan ekstrak
etanol Gynura
procumbens selama
24 jam menggunakan kepekatan
3.9 μg/mL hingga
500 μg/mL, makrofaj
RAW
264.7 menunjukkan kebolehhidupan
sel lebih
daripada 90%. Perencatan paras
penghasilan NO yang signifikan
telah dikenal
pasti dalam sel
RAW
264.7 yang dirangsang
oleh LPS bersama
pra-rawatan 250 μg/mL
ekstrak etanol
Gynura procumbens (p<0.05)
manakala semua
kepekatan ekstrak etanol Gynura procumbens yang dipilih menunjukkan tiada perubahan yang signifikan dalam penghasilan NO tanpa perangsangan LPS.
Pra-rawatan 250 μg/mL
ekstrak etanol
Gynura procumbens juga mendemonstrasikan perencatan ekspresi protein
iNOS yang signifikan dalam sel RAW 264.7
yang dirangsang oleh
LPS
(p<0.05). Kesimpulannya,
kajian ini menunjukkan bahawa ekstrak etanol Gynura procumbens mempamerkan
potensi anti-radang melalui perencatan penghasilan NO dan
ekspresi protein iNOS
dalam makrofaj yang dirangsang oleh LPS,
menunjukkan bahawa tumbuhan ini boleh
dikaji secara
lebih lanjut tentang
kegunaannya yang bermanfaat
dalam gangguan
keradangan.
Kata kunci: Gynura procumbens; lipopolisakarida; makrofaj;
nitrik oksida; radang;
sintase nitrik oksida teraruh
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*Corresponding author; email: drnorsyahida@ukm.edu.my
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