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PRF及所含三因子对大鼠脂肪干细胞迁移的影响

Effects of PRF and released three growth factors on migration of rat adipose tissue-derived stem cells
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摘要 目的 :研究富血小板纤维蛋白(platelet-rich fibrin,PRF)及所含三因子TGF-β1、PDGF-AB、VEGF对培养的大鼠脂肪干细胞(adipose tissue-derived stem cell,ADSCs)迁移的影响,并初步探讨其机制。方法:无菌切除SD大鼠腹股沟处的白色脂肪组织,采用酶消化法原代培养SD大鼠ADSCs,多向诱导分化法鉴定ADSCs,一次离心法提取制备PRF膜。采用划痕实验和Transwell实验检测细胞的迁移能力,实时荧光定量PCR分析迁移相关因子MT1-MMP和MMP-2 m RNA的表达情况。采用SPSS13.0软件包对数据进行统计学分析。结果 :Transwell实验显示,PRF组的迁移细胞数显著高于阴性对照组(P<0.05)和抑制剂组(P<0.05);不同浓度的TGFβ1、PDGF-AB、VEGF组的组内迁移细胞数的差异显著(P﹤0.05)。经PCR检测,PRF组细胞基因MMP2和MT1-MMP较对照组的表达显著上调(P<0.05),TGF-β1、PDGF-AB和VEGF的细胞基因MMP2和MT1-MMP较对照组表达均上调,组间差异显著(P<0.05)。结论 :PRF促进了ADSCs的迁移,PRF所分泌的三因子均可不同程度地促进ADSCs的迁移,并呈一定的量-效关系,其迁移能力的增加可能与MT1-MMP和MMP-2 m RNA的上调密切相关。 PURPOSE： To analyze the effects of PRF and released three growth factors on migration of rat adipose tissuederived stem cells and to investigate the mechanism of migration.METHODS： The inguinal adipose tissue of rat was excised at aseptic condition to obtain primary ADSCs by enzyme digestion.Multi-directional differentiation was used to identify the ADSCs.PRF membrane was acquired through one time centrifuge.The cell migration was examined by Transwell assay and wound healing assay.The m RNA expression of MMP2 and MT1-MMP was tested by real-time PCR.Statistical analysis was performed using SPSS 13.0 software package.RESULTS： Cell migration test showed that the migration of rat ADSCs in PRF group were significantly higher than those in the negative group（P 〈0.05） and inhibitor group（P〈0.05）.The ADSCs migration effects in three growth factors group at different concentrations showed significant difference（P〈0.05）.Real-time PCR showed that gene expressions of MMP2 and MT1-MMP were significantly higher in PRF group than control group（P〈0.05）.PCR showed that gene expressions of MMP2 and MT1-MMP were significantly higher in three growth factors group than control group（P 〈0.05）.CONCLUTIONS： PRF and three growth factors consistently enhanced the migration of rat ADSCs in a dose-response manner.The migration increase of rat ADSCs may be associated with the up-regulation of MMP2 and MT1-MMP gene expression.
作者 高洁 王明国 杨帅 李秀梅 杨世茂 李雪
机构地区 解放军济南军区第 济南市中心医院口腔科 济南市人民政府机关门诊部口腔科 潍坊医学院口腔医院
出处 《上海口腔医学》 CAS CSCD 2015年第6期667-673,共7页 Shanghai Journal of Stomatology
基金 山东省科技发展项目(2014GSF11804)~~
关键词 富血小板纤维蛋白 脂肪干细胞 细胞迁移 转化生长因子 血小板源性生长因子AB 血管内皮细胞生长因子 PRF ADSCs Migration TGF-β1 PDGF-AB VEGF
分类号 R783 [医药卫生—口腔医学]
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参考文献16

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二级参考文献28

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上海口腔医学
2015年 第6期

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