HIF-1α基因修饰的牙髓干细胞成血管的体外研究  

Dental pulp stem cells modified by HIF-1α can differentiate into blood vessels

作　　者：付洪海[1] 李鹏翀[2] 赵莉[2] 左金华[1] 

机构地区：[1]滨州医学院附属医院口腔颌面外科,山东滨州256603 [2]滨州市人民医院口腔科,山东滨州256603

出　　处：《上海口腔医学》2015年第6期674-678,共5页Shanghai Journal of Stomatology

基　　金：滨州医学院科技计划项目(BY2012KJ43)~~

摘　　要：目的 :探讨低氧诱导因子1α(hypoxia inducible factor 1α,HIF-1α)基因对体外诱导牙髓干细胞(dental pulp stem cells,DPSCs)成血管分化方法并进行鉴定。方法 :取拔除的健康、完整的前磨牙标本20例,提取DPSCs细胞,采用Strol-1、CD146分子进行鉴定。根据DPSCs是否转染HIF-1α基因分为实验组和对照组,RT-PCR法测定HIF-1αm RNA表达,Western印迹检测培养1、4、7、14 d不同时间段HIF-1α及成血管相关因子VEGF、SDF-1、Ang-2及PDGF的表达情况。采用SPSS16.0软件包对数据进行统计学分析。结果 :倒置相差显微镜观察,DPSCs多数细胞呈圆形、椭圆形类,免疫荧光观察到Strol-1、CD146均呈绿色荧光。实验组HIF-1α蛋白、m RNA随着时间的延长,表达水平越来越高,差异显著(P<0.05)。实验组转染后1、4、7、14 d后与对照组相比,HIF-1α蛋白、m RNA显著增高(P<0.05)。对照组血管相关因子VEGF、SDF-1、Ang-2和PDGF随时间的延长,其表达水平无显著差异(P>0.05)。实验组VEGF、SDF-1、Ang-2和PDGF随培养时间延长,其表达水平逐渐升高,不同时间点间差异显著(P<0.05)。与对照组相比,转染后1、4、7、14 d差异显著(P<0.05)。结论:HIF-1α基因修饰DPSCs能够成功体外诱导其血管分化作用,为进一步血管形成研究奠定了基础。PURPOSE： To investigate the method of differentiating dental pulp stem cells（DPSCs） modified by HIF-1αinto blood vessels.METHODS： DPSCs were extracted from teeth samples from 20 patients and were identified by Strol-1and CD146.DPSCs were divided into experimental group and control group according to DPSCs were modified by HIF-1 αnot or.HIF-1α-m RNA expression was detected by RT-PCR.HIF-1α,VEGF,SDF-1,Ang-2 and PDGF expression were detected using Western blot in different time after culture for 1 d,4 d,7 d and 14 d.Statistical analysis was carried out with SPSS 16.0 software package.RESULTS： Most DPSCs appeared round,oval under phase-contrast microscopy.CD146 and Strol-1 showed green fluorescence.HIF-1α and HIF-1α-m RNA expression became higher with time passing and the difference was statistically significant（P〈 0.05）.Compared with the control group,HIF-1α protein and m RNA increased obviously in the experimental group 1d,4d,7d and 14 d after transfection,and the difference was statistically significant（P〈0.05）.The level of VEGF,SDF-1,Ang-2 and PDGF in the control group was changed unconspicuously,and the expression was not different at different times（P〉 0.05）.The level of VEGF,SDF-1,Ang-2 and PDGF in the exprimental group increased,and the difference was statistically significant between different time points（ P 〈0.05）.Compared with the control group,the level of VEGF,SDF-1,Ang-2 and PDGF in the experimental group was higher 1 d,4 d,7 d and 14 d after transfection,respectively,and the difference was statistically significant（P〈0.05）.CONCLUSIONS：DPSCs modified by HIF-1α gene can successfully induce vascular differentiation in vitro,which provides foundation for further angioplasty.

关 键 词：低氧诱导因子1Α 牙髓干细胞 基因修饰 血管分化 

分 类 号：R78[医药卫生—口腔医学]