HGF对人股动脉内皮细胞中Notch1和Dll4基因转录的调节作用

Regulation of Dll4 and Notch1 genetic transcription by HGF in human femoral artery endothelial cells

目的:探讨肝细胞生长因子(HGF)对人股动脉内皮细胞(HFAECs)中Notch1和Dll4基因转录的调节作用,为进一步探讨携带HGF基因的重组腺病毒(Ad-HGF)促动脉血管形成机制奠定基础.方法:用不同感染强度(MOI)的携带绿色荧光蛋白基因的重组腺病毒(Ad-GFP)感染HFAECs后,通过流式细胞仪和MTT法选择最佳MOI,再将Ad-HGF以最佳的MOI感染HFAECs,48h后收集细胞上清,用ELISA法检测HGF的表达水平,同时提取总RNA,通过反转录-PCR(RT-PCR)来检测Notch1和Dll4基因的转录水平.以上均以感染Ad-GFP的细胞作为对照.结果:与对照组比较,Ad-HGF感染细胞后,上清中明显有HGF表达,并且观察到Notch1和Dll4基因转录水平均上调.结论:Ad-HGF促动脉形成机制可能与Notch信号通路有一定的关联性.

AIM： To investigate the regulation of Dll4 and Notch1 genetic transcription mediated by hepatocyte growth factor （HGF） in human femoral artery endothelial cells （ HFAECs ） , and to establish the basis for further investigating the mechanism of HGF promoting arteriogenesis. METHODS： The optimal multiplicity of infection （MOI） was determined by MTr and flow cytometry, after HFAECs were infected with Ad-GFP at the different MOI. Then HFAECs were infected with Ad-HGF at the optimal MOI, HGF in the supernatant was detected by ELISA at 48 h after infecting, meanwhile, the transcription of Notchl and Dll4 genes was detected by reverse transcription-PCR（RT-PCR）. Ad-GFP-infected HFAECs were used as negative controls. RESULTS： Compared to the control group, HGF was obviously detected in the supematant of Ad-HGF-infected HFAECs,and the up-regulation of Dll4 and Notchl genes transcription was observed. CONCLUSION： The mechanism of arteriogenesis promoted by HGF might be related with Notch signaling pathway.