中药杜仲补骨脂对去势鼠成骨细胞增殖以及MMP3/OPN通路蛋白表达的作用  

Effects of Eucommia Ulmoides and Fructus Psoraleae on osteoblast proliferation and expression of MMP3/OPN pathway proteins in ovariectomized rats

作　　者：王度 戴燚[2] 范彦博[2] 罗锟[2] 

机构地区：[1]武汉市第三医院,湖北武汉430060 [2]武汉市中医医院,湖北武汉430014

出　　处：《中国医院药学杂志》2016年第8期620-624,共5页Chinese Journal of Hospital Pharmacy

基　　金：湖北省卫生厅青年科技人才项目(编号:QJX2010-48);武汉市科技局应用基础研究计划项目(编号:2015060101010061);武汉市中青年医学骨干人才项目(编号:2014-41);武汉市青年科技晨光计划项目(编号:200950431215);武汉市卫生局临床医学项目(编号:WZ10C02)

摘　　要：目的:探讨中药杜仲补骨脂干预去势鼠成骨细胞增殖和MMP3-OPN-MAPK通路蛋白表达及其作用机制。方法:将10只饲养12周的去势鼠处死,选取股骨胫骨骨髓,进行体外成骨细胞传代培养。将含杜仲补骨脂药对2 g生药的药液喂养20只SD大鼠,随机均分为空白对照(A)组(给纯化水1 mL),低浓度(B)组(给药0.5 mL),中浓度(C)组(给药1 mL),高浓度(D)组(给药2 mL),提取各组处死大鼠含药血清干预成骨细胞,运用MTT法测定成骨细胞增殖能力,RT-PCR法测定通路蛋白MMP3、OPN和MAPK的mRNA水平。结果:(1)术后12周模型组大鼠腰椎及股骨骨密度为(0.163±0.012)g·cm^(-1)和(0.152±0.013)g·cm^(-1),假手术组为(0.326±0.015)g·cm^(-1)和(0.295±0.006)g·cm^(-1),两组有明显差异(P<0.05),表明大鼠骨质疏松的模型建立成功。(2)成骨细胞增殖中B至D组均使MTT法检测的光吸收值升高,较A组显著性增加(P<0.01和P<0.05),增加幅度与组别关系近似斜线,以D组促增殖作用最显著,增值率达到131%。(3)D组中MMP3含量明显低于其余各组(P<0.01),OPN和MAPK含量高于其余各组(P<0.05)。同时,B、C、D是随着复方含药血清依次增加的三组,B组中MMP3表达最多,其余组含量依次下降,三组中OPN含量基本依次增加,而MAPK含量在含药血清药物添加后明显增加,不同于A组(P<0.01)。结论:中药杜仲补骨脂药对含药血清可以促进去势鼠成骨细胞的增殖并且通过MMP3-OPNMAPK通路激活骨重建。OBJECTIVE To investigate expression of MMP3-OPN-MAPK pathway proteins in osteoblasts of ovariectomized rats intervened by Eucommia Ulmoides and Fructus Psoraleae,and the underlying mechanism.METHODS Ten rats after 12 weeks of feeding were sacrificed and bone marrow of femur was collected for cell passage.Twenty SD rats fed with drug liquid containing 2 g crude drug of Eucommia Ulmoides and Fructus Psoraleae were randomly divided into high concentration group（group A,2 mL of drug liquid）,middle concentration group（group B,1 mL）,low concentration group（group C,0.5 mL）and blank control group（group D,1 mL of distilled water）.Then rats were sacrificed to extract drug containing serum.MTT assay was used to measure proliferation of osteoblasts,RT-PCR to determine pathway MMP3,OPN and MAPK mRNA levels.RESULTS Bone densities of lumbar spine and femoral bone were（0.163±0.012）g·cm-1 and（0.152±0.013）g·cm-1 in model group 12 weeks after operation,（0.326±0.015）g·cm-1 and（0.295±0.006）g·cm-1 in sham operation group,and were significantly different between the two groups（P〈0.05）,suggesting osteoporosis model was successfully established.Light absorption determined by MTT was increased in group B to D,significantly higher than that in group A（P〈0.01,P〈0.05）.Relationship between increase amplitude and groups was like an oblique line.Group D had the most obvious proliferation promoting effects,which was increased by 131%.Content of MMP3 in group D was significantly lower than in other groups（P〈0.01）,and contents of OPN and MAPK were higher than in other groups（P〈0.05）.Of the three model groups,MMP3 expression was the highest in group B,and gradually declined in the remaining groups.OPN content increased dependently on drug concentration,while MAPK content significantly increased after adding drug containing serum,different from group A（P〈0.01）.CONCLUSION Eucommia Ulmoides and Fructus Psoraleae medicated serum can promote osteoblast proliferation and bone reconst

关 键 词：中药药对 成骨细胞 基质金属蛋白酶3 骨桥蛋白受体 信号通路 

分 类 号：R285.5[医药卫生—中药学]