摘要
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.
作者
周哲
宋海峰
曹天宇
刘涛
任建军
叶艳
张衍国
ZHOU Zhe;SONG Haifeng;CAO Tianyu;LIU Tao;REN Jianjun;YE Yan;ZHANG Yanguo(Department of Dermatology,Tangdu Hospital,the Air Force Medical University,Xi'an710038,China;Department of Dermatology,Xi'an North Hospital,Xi'an 710043,China)
出处
《中国皮肤性病学杂志》
CAS
CSCD
北大核心
2019年第8期869-874,共6页
The Chinese Journal of Dermatovenereology
基金
陕西省自然科学基金(2015SF164)
