Gaoli Xu, Yi Liu, Lili Wu, Qiutao Shi, Guang Huo, Zhiyuan Gu
School of Stomatology, Zhejiang Chinese Medical University, Hangzhou 310053, China;Dept. of Stomatology, Zhejiang Hospital, Hangzhou 310053, China.
School of Stomatology, Zhejiang Chinese Medical University, Hangzhou 310053, China;Dept. of Dental Implant and Prosthetics, University of Amsterdam, Amsterdam 1011-1109, Holland.
Hua Xi Kou Qiang Yi Xue Za Zhi. 2017 Jun 1;35(3):275-280. doi: 10.7518/hxkq.2017.03.009.
This study evaluates the biological effects of naringin (NAR) joint bone morphogenetic protein (BMP)-2 on the proliferation, alkaline phosphatase (ALP) activity, and expression of osteoblastogenic genes, such as Runt-related transcription factor 2 (Runx2), collagen Ⅰ (ColⅠ), ALP, and osteocalcin (OCN) of pre-osteoblasts.
Three different NAR concentrations (10, 100, and 1 000 μmol·L⁻¹) were applied, alone or combined with BMP-2(50 ng·mL⁻¹), to restore the osteoblastogenesis of pre-osteoblasts (MC3T3-E1 cell line). Cell numbers (proliferation) were evaluated at first, fourth, and seventh days by Alamar blue assay. ALP activity and the expression of osteoblastogenic genes, such as Runx2, ColⅠ, ALP, and OCN were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) at fourth and seventh day.
Stimulation by NAR alone and in combination with BMP-2 for 1 day and 4 days could promote cell proliferation, which peaked at a concentration of 100 μmol·L⁻¹ NAR combined with BMP-2 could promote cell proliferation significantly (P<0.05). Stimulation by NAR alone and in combination with BMP-2 for 4 and 7 days could promote ALP activity and bone-related gene(ALP, OCN, Runx2, ColⅠ) expression. ALP expression was significantly promoted after stimulation of 100 μmol·L⁻¹ NAR and BMP-2 (P<0.05).
CONCLUSIONS: NAR exhibits promising potential for improving MC3T3-E1 proliferation and differentiation, and appropriate concentrations of NAR and BMP-2 show synergistic effect. .
本研究评估柚皮苷（NAR）联合骨形态发生蛋白（BMP）-2对前成骨细胞增殖、碱性磷酸酶（ALP）活性以及成骨相关基因（如Runt相关转录因子2（Runx2）、Ⅰ型胶原（ColⅠ）、ALP和骨钙素（OCN））表达的生物学效应。
应用三种不同浓度的NAR（10、100和1000 μmol·L⁻¹），单独或与BMP-2（50 ng·mL⁻¹）联合使用，以恢复前成骨细胞（MC3T3-E1细胞系）的成骨能力。在第1、4和7天通过alamar蓝法评估细胞数量（增殖情况）。在第4天和第7天通过定量实时聚合酶链反应（qRT-PCR）分析ALP活性以及成骨相关基因（如Runx2、ColⅠ、ALP和OCN）的表达。
单独使用NAR以及NAR与BMP-2联合刺激1天和4天均可促进细胞增殖，在NAR浓度为100 μmol·L⁻¹时达到峰值，与BMP-2联合时可显著促进细胞增殖（P<0.05）。单独使用NAR以及NAR与BMP-2联合刺激4天和7天均可促进ALP活性和成骨相关基因（ALP、OCN、Runx2、ColⅠ）的表达。100 μmol·L⁻¹ NAR和BMP-2刺激后ALP表达显著增加（P<0.05）。
NAR在改善MC3T3-E1细胞增殖和分化方面具有良好潜力，适当浓度的NAR和BMP-2显示出协同效应。
