地黄单体梓醇促成骨细胞分化的活性研究  

Study on the Activity of Catalpol from Rehmannia glutinosa to Promote the Osteogenic Differentiation

作　　者：李路易 梁冬雨[2] 易清清 沙爽 史俊峰 梁鹏晨 常庆 LI Luyi;LIANG Dongyu;YI Qingqing;SHA Shuang;SHI Junfeng;LIANG Pengchen;CHANG Qing(Graduate School of Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China;General Medical Education and Research Center of Jiading District Central Hospital Affiliated to Shanghai University of Medicine & Health Sciences, Shanghai 201800, China;Shanghai Key Laboratory for Molecular Imaging, Shanghai University of Medicine & Health Sciences, Shanghai 201318, China;Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China)

机构地区：[1]上海中医药大学研究生院,上海201203 [2]上海健康医学院附属嘉定区中心医院全科医学教育与研究中心,上海201800 [3]上海健康医学院分子影像学重点实验室,上海201318 [4]上海中医药大学,上海201203

出　　处：《医药导报》2020年第4期533-537,共5页Herald of Medicine

基　　金：国家自然科学基金资助项目(81670968);上海市自然科学基金资助项目(19ZR1444800)。

摘　　要：目的检测并探索梓醇影响小鼠成骨细胞前体细胞MC3T3-E1增殖及成骨分化的有效浓度。方法CCK8法检测不同时间点(1,3,6 d)梓醇在不同浓度下对MC3T3-E1细胞安全性和增殖的影响;实时荧光定量PCR检测成骨分化关键性基因Runx2,Bglap和Col1α1 mRNA的表达水平;不同时间点(4,8 d)给予不同浓度梓醇后氨基安替吡啉测酚法分析细胞上清中碱性磷酸酶(alkaline phosphatase,ALP)活性的变化,茜素红染色法(alizarin red staining,ARS)检测细胞矿化水平的变化。结果不同浓度的梓醇都没有促进MC3T3-E1细胞的增殖,即使达到4000 mg·L^-1也不会导致细胞死亡;500 mg·L^-1的梓醇浓度能够显著提高细胞ALP活性,促进细胞矿化,提高成骨分化标志性基因的表达水平。结论梓醇具有促小鼠成骨细胞前体细胞MC3T3-E1成骨分化和矿化的作用,且对细胞具有较宽的安全浓度范围。Objective To determine and explore the effective concentration of catalpol on the proliferation and osteogenic differentiation of mouse osteoblast precursor cells MC3T3-E1.Methods CCK8 method was used to determine the effects of catalpol on the safety and proliferation of MC3T3-E1 cells at different time points(1,3,6 days).Quantitative fluorescence real-time PCR was used to detect the mRNA expression levels of osteogenic differentiation key genes,such as Runx2,Bglap,and Col1α1.Aminoantipyrine-phenol method was used to analyze the alkaline phosphatase activity in cell culture supernatants at different time points(4 and 8 days).Alizarin red staining was used to detect the changes of cell mineralization levels.Results Catalpol at different concentrations did not promote the proliferation of MC3T3-E1 cells.Even when the concentration of catalpol reached 4000 mg·L^-1,it would not cause cell death.Catalpol at the concentration of 500 mg·L^-1 could significantly increase the activity of ALP,promote cell mineralization,and enhance the expression levels of osteogenic differentiation marker genes.Conclusion Catalpol had the effect of promoting osteogenic differentiation and mineralization on mouse osteoblast precursor cells MC3T3-E1,and had a wide range of safe concentration for the cells.

关 键 词：梓醇 成骨分化 骨质疏松 

分 类 号：R96[医药卫生—药理学]