摘要
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.
出处
《第四军医大学学报》
北大核心
2009年第20期2073-2076,共4页
Journal of the Fourth Military Medical University
基金
国家自然科学基金面上项目(30772572)
关键词
肝细胞生长因子
NOTCH信号通路
动脉生成
基因转录
hepatocyte growth factor ( HGF )
Notch signing pathway
arteriogenesis
genetic transcription