RT-PCR检测釉基质蛋白对大鼠骨髓基质细胞成骨相关基因表达影响的研究  

Effect of enamel matrix proteins on expression of bone formation-related genes of rat BMSCs were detected by RT-PCR

作　　者：吴飞俊[1] 赵芳[1] 裴路[1] 

机构地区：[1]哈尔滨医科大学附属第二医院口腔科,哈尔滨550012

出　　处：《口腔医学》2013年第6期366-370,共5页Stomatology

基　　金：黑龙江省自然科学基金(D2007-10)

摘　　要：目的研究釉基质蛋白(enamel matrix proteins,EMPs)对体外培养大鼠骨髓基质细胞(bone marrow stromal cells,BM-SCs)成骨相关基因表达的影响。方法采用乙酸法从猪恒牙胚中提取釉基质蛋白,取3周龄雄性SD大鼠胫骨和股骨骨髓体外培养,取生长良好的第3代细胞加入不同浓度的EMPs(0、25、50、100、200 mg/L)进行培养,MTT法检测BMSCs的增殖,用RT-PCR测定成骨相关基因骨涎蛋白(bone sialoprotein,BSP),骨桥蛋白(osteopontin,OPN)和Ⅰ型胶原(collagen typeⅠ,COL-Ⅰ)的表达。结果实验组的细胞增殖大于对照组(P<0.05),并且EMPs的浓度在100 mg/L时增殖作用最强;实验组骨相关基因(BSP、COL-Ⅰ)mRNA的表达高于对照组,并且具有浓度依赖性,但是实验组与对照组OPN mRNA的表达没有区别。结论 EMPs通过调节成骨相关基因的表达而促进大鼠BMSCs向成骨细胞分化。Objective To study the effects of EMPs on expression of bone formation-related gene of rat bone marrow stromal cells culutured in vitvo by RT-PCR.Methods Enamel matrix proteins were extracted from porcine permanent tooth buds by acidic extraction.Marrow of tibiae and femurs from 3-week-old male SD rats was obtained and cultured in vitro.Well-grown cells at passage 3 were cultured with different concentrations of EMPs（0,25,50,100,200 mg/L）.MTT assay was performed to evaluate the proliferation of BMSCs.The expression of bone formation-related genes BSP,OPN and COL-Ⅰwas detected by RT-PCR.Results Cell proliferation in experimental group was higher than that in control group（P〈0.05）,and the proliferation was highest when concentration of EMPs was 100 mg/L.The mRNA expression of bone-related genes（BSP、COL-Ⅰ）in experimental group was higher than that in control group,and was in a concentration dependent manner.But,there was no difference between the control group and the experimental group in mRNA expression of OPN gene.Conclusions EMPs enhance osteoblastic differentiation of rat BMSCs by regulation of bone formation-related gene expression.

关 键 词：釉基质蛋白 骨髓基质细胞 基因表达 成骨相关基因 

分 类 号：R349.64[医药卫生—基础医学]