血小板衍生生长因子对人脐带间充质干细胞胞外基质的影响

Effects of the platelet-derived growth factor on proliferation and extracellular matrix of human umbilical cord mesenchymal stem cells

目的探讨血小板衍生生长因子(PDGF)对人脐带间充质干细胞(UCMSCs)增殖和细胞外基质(ECM)表达的影响及其可能的作用机制。方法体外传代培养UCMSCs,分为对照组和实验组,其中对照组不加PDGF,实验组分别加入不同质量浓度(2、5、10、20、50μg/L)的PDGF。加入PDGF12h后,用qRT-PCR法检测ECM基因和凋亡相关基因(Bcl-2、Bax)的表达;加入PDGF24h后,用四甲基偶氮唑(MTT)法检测细胞增殖。结果与对照组比较,随着PDGF质量浓度的增加,反映细胞代谢活性的OD值逐渐增加,且PDGF质量浓度在20μg/L时达到最高值,但PDGF质量浓度为50μg/L时OD值反而降低;PDGF质量浓度为2μg/L时,ECM表达和Bcl-2/Bax比值较对照组有所降低,随着PDGF质量浓度的增加,ECM的表达和Bcl-2/Bax比值呈现先增高后降低的趋势,且在10μg/L时达到最高值。结论 PDGF可以促进脐带间充质干细胞的增殖,同时能促进其ECM的表达,还能提高细胞耐受凋亡的能力,最佳作用质量浓度为10μg/L。

Objective To investigate the effects of the platelet-derived growth factor（PDGF） on the proliferation,extracellular matrix synthesis and its probable mechanism in human umbilical cord mesenchymal stem cells（UCMSCs） .Methods Cultured UCMSCs were divided into 6 groups：the control group,and the PDGF-treated groups（2,5,10,20 and 50 μg/L） .Morphological changes of the cells were observed under an inverted microscope.MTT assay was used to detect the PDGF effects on cell proliferation.Expressions of ECM mRNA and apoptosis-related genes（Bcl-2,Bax） were detected by qRT-PCR.Results With an increased concentration of PDGF,UCMSCs proliferation was gradually promoted compared with the control group.20 μg/L PDGF greatly promoted UCMSCs proliferation,however,50 μg/L showed inhibitive effects.Expressions of ECM mRNA and ap-optosis-related factor （Bcl-2/Bax） were down-regulated in the 2 μg/L PDGF group but up-regulated when PDGF≥5 μg/L.10 μg/L PDGF greatly up-regulated ECM mRNA expression and apoptosis-related factor （Bcl-2/Bax） .Conclusion PDGF can promote the proliferation of UCMSCs,stimulate ECM synthesis and improve the ability to withstand apoptosis of UCMSCs in vitro.10 μg/L PDGF was the most effective concentration.