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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。
Objective: To investigate the effects of platelet-rich fibrin (PRF) and three factors TGF-β1, PDGF-AB and VEGF on the migration of cultured adipose tissue-derived stem cells (ADSCs) And preliminary discussion of the mechanism. Methods: Adipose tissue was used to excise white adipose tissue in the groin of AD rats. ADSCs were primarily cultured by enzymatic digestion method. ADSCs were identified by multidirectional induction and differentiation method. PRF membrane was prepared by centrifugation. Scratch assay and Transwell assay were used to detect the cell migration ability. The expression of MT1-MMP and MMP-2 mRNA were analyzed by real-time fluorescence quantitative PCR. SPSS13.0 software package for statistical analysis of the data. Results: Transwell assay showed that the number of migrated cells in PRF group was significantly higher than that in negative control group and inhibitor group (P <0.05). The numbers of migrated cells in TGFβ1, PDGF-AB and VEGF groups The difference was significant (P <0.05). The expression of MMP2 and MT1-MMP in PRF group was significantly up-regulated compared with the control group (P <0.05). The expressions of MMP2 and MT1-MMP in TGF-β1, PDGF-AB and VEGF were up-regulated compared with the control group There was significant difference between groups (P <0.05). CONCLUSION: PRF promotes the migration of ADSCs. The three factors secreted by PRF can promote the migration of ADSCs to some extent, and show a certain dose-effect relationship. The increase of migration capacity may be related to the increase of MT1-MMP and MMP-2 mRNA The increase is closely related.