原儿茶酸促进人脂肪干细胞体外迁移研究

Research on promotion of human adipose tissue-derived stromal cells migration by protocatechuic acid in vitro

研究了益智仁中提取的原儿茶酸对人脂肪干细胞(hADSCs)体外迁移的影响.实验采用明胶包被的Transwell细胞培养池,观察到适宜浓度的原儿茶酸可显著促进人脂肪干细胞体外迁移,其作用呈现明显剂量依赖性.同时,1.5 mmol/L原儿茶酸促进hADSCs迁移的作用最强且呈现时间依赖性.RT-PCR和荧光定量PCR结果显示,经原儿茶酸处理后,人脂肪干细胞膜型基质金属蛋白酶-1(MT1-MMP)的表达明显升高.酶谱分析发现,经原儿茶酸处理后,干细胞的活性型基质金属蛋白酶-2(MMP-2)增加.抗体阻断实验显示,细胞迁移能力的增强与MT1-MMP的表达升高和MMP-2的活化增强有关.最后,人脂肪干细胞鉴定结果表明,经原儿茶酸处理后的人脂肪干细胞仍保持间充质干细胞多分化潜能的特性.因此,原儿茶酸有可能在脂肪干细胞移植的治疗中发挥作用.

The effect of protocatechuic acid （PCA） from Alpinia oxyphylla on the celt migration of human adipose tissue-derived stromal cells （hADSCs） was studied. It was found that PCA could promote the migration capacity of hADSCs through Transwell coated with gelatin in vitro. PCA enhanced the cell migration in a dose-dependent manner. Meanwhile, the optimal effect of PCA was observed at 1.5 mmol/L, and the cell migration rate increased in a time-dependent manner with treatment of PCA （1.5 mmol/L）. RT-PCR and quantitative RT-PCR analysis revealed the elevation of membrane-type matrix metalloproteinase-1 （MT1-MMP） mRNA expression in PCA-treated hADSCs. In cell supernatants of PCA-treated hADSCs, the active matrix metalloproteinase-2 （MMP-2） increased compared with control cells, as detected by zymography. The results of antibody blocking experiments show that the promotion of cell migration by PCA is effectively and obviously inhibited by MT1-MMP or MMP-2 antibodies. Furthermore, after PCA treatment, hADSCs still retain their functional characteristics of multipotential mesenchymal progenitors. The cell migration enhancement of hADSCs with PCA suggests the possibility that PCA may be useful in hADSCs-mediated therapy.