PRF及所含三因子对大鼠脂肪干细胞迁移的影响

上海口腔医学 ›› 2015, Vol. 24 ›› Issue (6): 667-673.

PRF及所含三因子对大鼠脂肪干细胞迁移的影响

高洁¹，王明国²，杨帅²，李秀梅³，杨世茂²，李雪⁴

1.解放军济南军区第401医院口腔科，山东青岛266071；
2.济南市中心医院口腔科，山东济南250013；
3.济南市人民政府机关门诊部口腔科，山东济南250099；
4.潍坊医学院口腔医院，山东潍坊261053

收稿日期:2015-05-19 出版日期:2015-12-25 发布日期:2016-01-04
通讯作者: 王明国，Tel:0531-85695171，E-mail：wmgsh@163.com E-mail:gaojierizhao@126.com
作者简介:高洁1987-，女，硕士，住院医师
基金资助:
Supported by Science and Technology Development Program of Shandong Province (2014GSF11804).

Effects of PRF and released three growth factors on migration of rat adipose tissue-derived stem cells

GAO Jie¹, WANG Ming-guo², YANG Shuai², LI Xiu-mei³, YANG Shi-mao², LI Xue⁴.

1.Department of Stomatology, the 401st Hospital of Jinan Military Area, PLA.Qingdao 266071;
2.Department of Stomatology, Jinan Central Hospital. Jinan 250013;
3.Department of Stomatology, Jinan Municipal People’s Government. Jinan 250099;
4.Hospital of Stomatology, Weifang Medical University. Weifang 261053, Shandong Province, China

Received:2015-05-19 Online:2015-12-25 Published:2016-01-04
Contact: 山东省科技发展项目（2014GSF11804） E-mail:gaojierizhao@126.com

摘要/Abstract

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 mRNA 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 mRNA的上调密切相关。

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

Abstract: PURPOSE To analyze the effects of PRF and released three growth factors on migration of rat adipose tissue-derived 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 mRNA 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.

Key words: PRF, ADSCs, Migration, TGF-β1, PDGF-AB, VEGF

中图分类号:

R782

引用本文

高洁，王明国，杨帅，李秀梅，杨世茂，李雪. PRF及所含三因子对大鼠脂肪干细胞迁移的影响[J]. 上海口腔医学, 2015, 24(6): 667-673.

GAO Jie, WANG Ming-guo, YANG Shuai, LI Xiu-mei, YANG Shi-mao, LI Xue.. Effects of PRF and released three growth factors on migration of rat adipose tissue-derived stem cells[J]. Shanghai Journal of Stomatology, 2015, 24(6): 667-673.

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