γ分泌酶抑制剂对肝血窦内皮细胞的影响  

The Impact of Gamma Secretase Inhibitors to Liver Sinusoidal Endothelial Cells

作　　者：金超[1] 任凯夕[2] 晏贤春[2] 马鹏飞[1] 郭丰成[1] 窦科峰[1] 

机构地区：[1]第四军医大学西京医院肝胆外科,陕西西安710032 [2]第四军医大学基础部遗传发育教研室,陕西西安710032

出　　处：《现代生物医学进展》2013年第16期3019-3023,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81030010)

摘　　要：目的:在建立高纯度小鼠肝血窦内皮细胞的体外培养的基础上研究γ分泌酶抑制剂(DAPT)对肝血窦内皮细胞活性的影响。方法:首先通过胶原酶灌注消化、percoll梯度离心和选择性贴壁分离得到高纯度、可在体外条件下培养的肝血窦内皮细胞,其次用不同浓度的DAPT(15μmol/L、45μmol/L、75μmol/L)处理细胞,然后通过MTT检测细胞增殖情况、Real time PCR检测相关分子改变。结果:在体外条件下DAPT对肝血窦内皮细胞的增殖起到促进作用,这种促进作用随着DAPT浓度的增加相应的增加;DAPT能够导致肝血窦内皮细胞Notch信号下游分子Hes1表达下调,VEGF信号中VEGFR1表达下调,VEGFR2表达上调。结论:γ分泌酶抑制剂(DAPT)通过抑制肝血窦内皮细胞Notch信号,引起肝血窦内皮细胞表面VEGFR1表达下调,VEGFR2表达上调显著增加肝血窦内皮细胞的活性。Objective： Our study focused on the impact of gamma secretase inhibitors（DAPT） to liver sinusoidal endothelial cells（LSECs） on the basis of the establishment of high-purified mouse liver sinusoidal endothelial cells in vitro.Methods： We obtained high-purified liver sinusoidal endothelial cells in vitro by collagenase perfusion,percoll gradient centrifugation and selective adherence separation.The cells were treated by DAPT with different concentrations（15 mol/L,45 mol/L,75 mol/L）.Cell proliferation was detected by MTT and molecular changes was detected by Real-time PCR.Results： DAPT promoted sinusoidal endothelial cell proliferation in vitro,which was correspondent to the increasing concentrations of DAPT.Notch signaling downstream molecules Hes1 of liver sinusoidal endothelial cells was down-regulated by DAPT.Expression of VEGFR1 was down-regulated and expression of VEGFR2 was up-regulated by DAPT.Conclusions： Gamma secretase inhibitors（DAPT） inhibited Notch signaling of liver sinusoidal endothelial cells,causing down-regulation of liver sinusoidal endothelial cell surface VEGFR1 and up-regulation of VEGFR2,and increased the activity of the liver sinusoidal endothelial cells.

关 键 词：DAPT 肝血窦内皮细胞 NOTCH信号 

分 类 号：Q95-3[生物学—动物学] R657.3[医药卫生—外科学]