Sains Malaysiana 49(6)(2020): 1389-1400

http://dx.doi.org/10.17576/jsm-2020-4906-17

Enhancing Effect of Vitexin on Osteogenic Activity of Murine Pre-Osteoblastic MC3T3-E1 Cells

(Kesan Peningkatan Viteksin terhadap Aktiviti Osteogenik Sel MC3T3-E1 Pra-Osteoblastik Murin)

XIAOHAN YUAN¹, HAIYAN HAN², ZHAOHUI LUO³, QIUXUE WANG², PEIJIA TANG², ZHIHUI ZHANG², YUJIE FU² & CHENGBO GU^1,2*

¹Key Laboratory of Agricultural Microbiology of Heilongjiang Provincial Science and Technology Department, Northeast Agricultural University, Harbin, 150030, China

²Key Laboratory of Forest Plant Ecology, Ministry of Education, Northeast Forestry University, Harbin, 150040, China

³Fankou Lead-Zinc Mine Staff Hospital, Shenzhen Zhongjin Lingnan Nonfemet Company Limited, Renhua, 512325, China

Received: 17 September 2019/Accepted: 14 February 2020

ABSTRACT

Vitexin (5,7,4-trihydroxyflavone-8-glucoside), a natural flavone present in a variety of plants, is well known for its rich pharmacological properties. However, its osteogenic activity remains unclear to date. The purpose of this study was to explore the effects of vitexin on osteogenic activity in murine pre-osteoblastic MC3T3-E1 cells using the MTT assay for cell proliferation, alkaline phosphatase (ALP) activity assay for cell differentiation, and Von Kossa staining for cell mineralization. Quantitative real-time PCR was used for the detection of osteocalcin (OCN) mRNA expression in cells. Furthermore, effects of vitexin on the differentiation and matrix mineralization of dexamethasone (DEX)-suppressed cells was also investigated. The results showed vitexin could significantly enhance cell proliferation in a low concentration range of 10-10-10-6 μg mL-1. ALP activity was significantly increased after the cells were treated with vitexin at 10-8 and 10-6 μg mL-1. The expression levels of the osteogenic OCN gene in cells treated with vitexin at 10-6, 10-8, and 10-10 μg mL-1 were improved by 3.1-fold, 5.8-fold, and 4.2-fold over the control, respectively. Additionally, vitexin (10-8 μg mL-1) significantly alleviated the inhibitory effect of osteoblast differentiation and mineralization induced by DEX. Collectively, our findings suggest vitexin could enhance cell proliferation and osteogenic differentiation of MC3T3-E1 cells, as well as rescue the inhibitory effect of cell differentiation and matrix mineralization induced by DEX. Therefore, vitexin may be useful as a promising therapeutic agent for bone disease and plays an important role in the prevention of glucocorticoid-induced osteoporosis.

Keywords: Anti-osteoporosis; dexamethasone; MC3T3-E1 cells; osteogenic activity; vitexin

ABSTRAK

Viteksin (5,7,4-trihidroksiflavon-8-glucosida), merupakan flavon semula jadi yang terdapat dalam pelbagai jenis tumbuhan juga terkenal dengan khasiat farmakologinya yang tinggi. Walau bagaimanapun, aktiviti osteogeniknya masih belum jelas sehingga kini. Tujuan kajian ini adalah untuk mengkaji kesan viteksin terhadap aktiviti osteogenik pada sel MC3T3-E1 pra-osteoblas murin menggunakan asai MTT untuk proliferasi sel, asai aktiviti alkali fosfatase (ALP) untuk pembezaan sel dan pewarnaan Von Kossa untuk mineralisasi sel. PCR masa nyata kuantitatif digunakan untuk mengesan ekspresi mRNA osteokalsin (OCN) dalam sel. Selanjutnya, kesan viteksin terhadap pembezaan dan mineralisasi matriks deksametason (DEX) sel yang ditindas juga dikaji. Hasil kajian menunjukkan viteksin dapat meningkatkan proliferasi sel secara signifikan dalam julat kepekatan yang rendah iaitu 10-10-10-6 μg mL-1. Aktiviti ALP meningkat dengan ketara setelah sel dirawat dengan viteksin pada kepekatan 10-8 dan 10-6 μg mL-1. Tahap ekspresi gen OCN osteogenik dalam sel yang dirawat dengan viteksin pada 10-6, 10-8 dan 10-10 μg mL-1 masing-masing meningkat sebanyak 3.1, 5.8 dan 4.2 kali ganda daripada kawalan. Selain itu, viteksin dengan kepekatan 10-8 μg mL-1 juga didapati meningkatkan kesan perencatan pembezaan osteoblas dan mineralisasi yang diaruh oleh DEX. Secara kolektifnya, hasil kajian ini menunjukkan bahawa viteksin dapat meningkatkan proliferasi sel dan pembezaan osteogenik sel MC3T3-E1, serta melindungi daripada kesan perencatan pembezaan osteoblas dan mineralisasi yang diaruh oleh DEX. Oleh itu, viteksin berpotensi digunakan sebagai agen terapi yang baik bagi merawat penyakit tulang dan memainkan peranan penting dalam pencegahan osteoporosis yang disebabkan oleh glukokortikoid.

Kata kunci: Aktiviti osteogenik; anti-osteoporosis; deksametason; sel MC3T3-E1; viteksin

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*Corresponding author; email: dilisatis@163.com