牵张力作用下瘢痕疙瘩间充质干细胞通过旁分泌作用促进血管生成

Keloid-derived Mesenchymal Stem Cells Under Tension Promote Angiogenesis Through Paracrine Mechanisms

目的研究牵张力作用下瘢痕疙瘩间充质干细胞(keloid-derived mesenchymal stem cells,KD-MSCs)培养上清对人脐静脉血管内皮细胞(human umbilical vein endothelial cells,HUVECs)生物活性的影响。方法采用CCK-8法、平板划痕实验及Transwell小室法检测牵张力作用下KD-MSCs培养上清对HUVECs的增殖、细胞迁移情况的影响,并通过RT-PCR检测其整合素(integrin)αv、β3、VEGF、TGF-β、PDGF基因表达的变化。结果牵张力作用下KD-MSCs培养上清刺激HUVECs后其细胞生物活性有明显差异;CCK-8分析结果显示牵张力组HUVECs的增殖高于无牵张力组和空白对照组(P<0.05);平板划痕实验和Transwell小室实验结果显示牵张力组HUVECs的迁移能力明显强于无牵张力组和空白对照组(P<0.05);同时RT-PCR检测结果表明牵张力组HUVECs中整合素αv、β3mRNA表达显著低于无牵张力组(P<0.05);VEGF mRNA表达显著高于无牵张力组(P<0.05);TGF-β、PDGF无明显变化。结论牵张力作用下KD-MSCs可能通过旁分泌促进HUVECs增殖和迁移,其中整合素αv、β3和VEGF参与该生物学过程从而可能在瘢痕疙瘩的发生发展中发挥作用。

Objective To investigate the effect of supernatant from keloid-derived mesenchymal stem cells(KD-MSCs)under tensile force on the biological activity of human umbilical vein endothelial cells(HUVECs)under tensile force.Methods Cell counting Kit-8(CCK-8)assay,the scratch test,and transwell migration assay were used to detect the effects of KD-MSCs supernatant on cell proliferation and migration of HUVECs.Real-time PCR was used to detect the mRNA expression of integrin αv,β3,VEGF,TGF-β,PDGF in HUVECs.Results There were significant differences in the biological activity of HUVECs under tensile force.CCK-8 assay showed that the proliferation ability of HUVECs in tension group was higher than in static group and control group(P<0.05).The scratch test and transwell migration assay demonstrated that the migration ability of HUVECs in tension group was higher than in static group and control group(P<0.05).The mRNA expression level of integrin αv and β3 was significantly decreased,while that of VEGF was significantly increased in tension group compared with static group(P<0.05).There were no significant differences in the mRNA expression levels of TGF-β and PDGF.Conclusion KD-MSCs stimulated with tensile force may promote the cell proliferation and migration of HUVECs through paracrine mechanisms.Furthermore,integrin αv,β3 and VEGF may participate in the occurrence and development of keloids.