上海口腔医学 ›› 2015, Vol. 24 ›› Issue (6): 674-678.

• 基础研究 • 上一篇    下一篇

HIF-1α基因修饰的牙髓干细胞成血管的体外研究

付洪海1,李鹏翀2,赵莉2,左金华1   

  1. 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
  • 收稿日期:2015-05-18 出版日期:2015-12-25 发布日期:2016-01-04
  • 通讯作者: 左金华,Tel:0543-3258687,E-mail:jinhua256603@163.com E-mail:renxinmiaoshu@hotmail.com
  • 作者简介:付洪海1983-,男,硕士,住院医师
  • 基金资助:
    Supported by Science and Technology Research Program of Binzhou Medical University(BY2012KJ43).

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

FU Hong-hai1, LI Peng-chong2, ZHAO Li2, ZUO Jin-hua1.   

  1. 1.Department of Oral and Maxillofacial Surgery, Affiliated Hospital of Binzhou Medical University. Binzhou 256603;
    2.Department of Stomatology, Binzhou People’s Hospital. Binzhou 256603, Shandong Province, China
  • Received:2015-05-18 Online:2015-12-25 Published:2016-01-04
  • Contact: 滨州医学院科技计划项目(BY2012KJ43) E-mail:renxinmiaoshu@hotmail.com

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-1α mRNA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、mRNA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、mRNA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。

关键词: 低氧诱导因子1α, 牙髓干细胞, 基因修饰, 血管分化

Abstract: PURPOSE To investigate the method of differentiating dental pulp stem cells (DPSCs) modified by HIF-1α into blood vessels. METHODS DPSCs were extracted from teeth samples from 20 patients and were identified by Strol-1 and CD146. DPSCs were divided into experimental group and control group according to DPSCs were modified by HIF-1α not or. HIF-1α-mRNA 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-mRNA expression became higher with time passing and the difference was statistically significant (P<0.05). Compared with the control group, HIF-1α protein and mRNA increased obviously in the experimental group 1d, 4d, 7d and 14d 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.

Key words: HIF-1α, DPSCs, Gene modification, Vessel differentiation

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