Young-Eun Cho; In-Sook Kwun

doi:10.4163/jnh.2018.51.4.316

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Cho and Kwun: Glycyrrhiza uralensis (licorice) extracts increase cell proliferation and bone marker enzyme alkaline phosphatase activity in osteoblastic MC3T3-E1 cells∗

Research Article

J Nutr Health 2018;51(4):316-322.

Published online: 20 January 2018

DOI: https://doi.org/10.4163/jnh.2018.51.4.316

Glycyrrhiza uralensis (licorice) extracts increase cell proliferation and bone marker enzyme alkaline phosphatase activity in osteoblastic MC3T3-E1 cells^∗

Young-Eun Cho, In-Sook Kwun^†

Department of Food Science and Nutrition, Andong National University, Andong, Gyeongbuk 36729, South Korea

^† To whom correspondence should be addressed. tel: +82-54-820-5917, E-mail: iskwun@andong.ac.kr

^∗ This work was supported by grants from by Andong National University (ANU) under 2016 ANU research fund.

Received 2 July 2018 Revised 29 July 2018 Accepted 1 August 2018

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Purpose

The Glycyrrhiza uralensis species (Leguminosae) as a medicinal biocompound, and one of its root components, isoliquritigenin (ISL), which is a flavonoid, has been reported to have anti-tumor activity in vitro and in vivo. However, its function in bone formation has not been studied yet. In this study, we tested the effect of Glycyrrhiza uralensis (ErLR) and baked Glycyrrhiza uralensis (EdLR) extracts on osteoblast proliferation, alkaline phosphatase (ALP) activity, and bone-related gene expression in osteoblastic MC3T3-E1 cells. Methods: MC3T3-E1 cells were cultured in various levels of ErLR (0, 5, 10, 15, 20 μg/mL), EdLR (0, 5, 10, 15, 20 μg/mL), or ISL (0, 5, 10, 15, 20 μM) in time sequences (1, 5, and 20 days). Also, isoliquritigenin (ISL) was tested for comparison to those two biocompound extracts. Results: MTT assay results showed that all three compounds (ErLR, EdLR, and ISL) increased osteoblastic-cell proliferation in a concentration-dependent manner for one day. In addition, both ErLR and EdLR compounds elevated the osteoblast proliferation for 5 or 20 days. Extracellular ALP activity was also increased as ErLR, EdLR, and ISL concentration increased at 20 days, which implies the positive effect of Glycyrrhiza species on osteoblast mineralization. The bone-related marker mRNAs were upregulated in the ErLR-treated osteoblastic MC3T3-E1 cells for 20 days. Bone-specific transcription factor Runx2 gene expression was also elevated in the ErLR- and EdLR-treated osteoblastic MC3T3-E1 cells for 20 days. Conclusion: These results demonstrated that Glycyrrhiza uralensis extracts may be useful for preventing osteoporosis by increasing cell proliferation, ALP activity, and bone-marker gene expression in osteoblastic cells.

Keywords: Glycyrrhiza uralensis, MC3T3-E1 cells, cell proliferation, ALP activity, Runx2

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Fig. 1.

Morphological characteristics of osteoblastic MC3T3-E1 cells treated with various ErLR, EdLR, and ISL compounds. Photographs were taken with a phase-contrast microscope at 100 × magnification.

Fig. 2.

Effect of ErLR (A), EdLR (B), and ISL (C) compounds on the proliferation of osteoblastic MC3T3-E1 cells. Significant differences between various treatments were found by one-way ANOVA. Labelled characters without a common letter represent significant differences from the other group(s). The values are presented as % of control at 0 h.

Fig. 3.

Effect of ErLR (A), EdLR (B), and ISL (C) compounds on the medium ALP activity of osteoblastic MC3T3-E1 cells. Significant difference between treatments were found by one-way ANOVA. Labelled characters without a common letter represent significant differences from the other group(s) at p < 0.05.

Fig. 4.

Gene expression of bone-marker genes (ALP and COLI) and transcription factor Runx2 in osteoblastic MC3T3-E1 cells treated with various ErLR, EdLR, and ISL compounds for 20 days. (A-B) mRNA transcription level of bone markers was measured using quantitative real-time PCR (qRT-PCR). Significant difference between treatments were found by one-way ANOVA. Labelled characters without a common letter represent significant differences from the other group(s).

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