上海交通大学学报(医学版)

• 论著(基础研究) • 上一篇    下一篇

大黄素通过BMP-9途径促进前体成骨细胞的分化

陈小静,胡 燕,张 爽,高艳虹   

  1. 上海交通大学 医学院附属新华医院老年医学科, 上海 200092
  • 出版日期:2014-06-28 发布日期:2014-06-30
  • 通讯作者: 高艳虹, 电子信箱: yhgao2010@yahoo.com。
  • 作者简介:陈小静(1987—), 女, 硕士生; 电子信箱: cxj061@163.com。
  • 基金资助:

    国家自然科学基金(81101360);上海市教委科研创新项目(12YZ053)

Emodin promotes differentiation of preosteoblast via BMP-9 signaling pathway

CHEN Xiao-jing, HU Yan, ZHANG Shuang, GAO Yan-hong   

  1. Department of Geriatrics, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China
  • Online:2014-06-28 Published:2014-06-30
  • Supported by:

    National Natural Science Foundation of China,81101360; Innovation Program of Shanghai Municipal Education Commission,12YZ053

摘要:

目的 研究大黄素对前体成骨细胞MC3T3-E1增殖与分化的影响及其作用机制。方法 采用不同浓度(0.05、0.1、0.5、2.0和5.0 μmol/L)大黄素处理MC3T3-E1细胞(给药组),以不加大黄素为对照组。分别采用MTT法检测细胞的增殖率;对硝基苯基磷酸二钠(PNPP)法定量检测碱性磷酸酶(ALP)活性;茜素红染色观察矿化结节数目;RT-PCR法检测骨形态发生蛋白(BMPs)途径相关信号分子的表达。采用BMPs抑制剂(Noggin)处理细胞后检测ALP活性,观察BMP-9在大黄素促成骨分化中的作用。结果 大黄素对MC3T3-E1细胞的增殖无影响,大黄素浓度为0.5~5.0 μmol/L时可促进细胞ALP表达及矿化结节形成,其中浓度为5 μmol/L时效果最显著。大黄素可增强细胞BMP-9的表达,与BMP-9信号途径相关的上下游信号分子mRNA的表达亦有相应改变;Noggin预处理后,大黄素促成骨分化的功能受到明显抑制。结论 大黄素可通过激活BMP-9信号通路促进前体成骨细胞的分化。

关键词: 大黄素, 前体成骨细胞, 分化, 骨形态发生蛋白9

Abstract:

Objective To explore the effects of emodin on proliferation and differentiation of MC3T3-E1 preosteoblasts and the possible mechanism. Methods The treated group contained MC3T3-E1 cells treated by emodin of different concentrations (0.05, 0.1, 0.5, 2.0, and 5.0 μmol/L) and the control group contained MC3T3-E1 cells not treated by emodin. The cell proliferation rate was detected by the MTT assay. The activity of ALP was determined by the quantitative PNPP method. The alizarin red staining was used to observe the number of mineralized nodules. The expression of signaling molecules relevant to BMP pathway was detected by the RT-PCR. The activity of ALP was measured and the effects of BMP-9 on the differentiation of osteoblasts were observed after cells were treated by Noggin, which was a BMP-specific inhibitor. Results Emodin had no effect on the proliferation of MC3T3-E1 cells. Emodin increased the expression of ALP and formation of mineralized nodules at the concentrations from 0.5 μmol/L to 5 μmol/L, especially at the concentration of 5 μmol/L. Emodin enhanced the expression of BMP-9 and expressions of upstream and downstream signaling molecules relevant to BMP-9 pathway also changed accordingly. The emodin-induced differentiation of osteoblasts was inhibited after being pretreated by Noggin. Conclusion Emodin can promote the differentiation of preosteoblasts by activating the signaling pathway of BMP-9.

Key words: emodin, preosteoblast, differentiation, bone morphogenetic protein 9