不同氧浓度对原代培养人早孕绒毛细胞滋养细胞血管内皮生长因子及其可溶性受体表达的影响

Expression of Vascular Endothelial Growth Factor and Soluble Fms-like Tyrosine Kinase Receptor 1 under Different Oxygen Condition in the Primary Cultured Cytotrophoblast Cells of Human

目的:探讨原代培养人早孕绒毛细胞滋养层细胞(CT)在缺氧时血管内皮生长因子(VEGF)和可溶性血管内皮生长因子受体-1(sFlt-1)的表达。方法:将经胰蛋白酶/DNA酶Ⅰ联合消化通过Percoll分离纯化得到的细胞滋养层细胞进行原代培养。采用ELISA法测定正常氧浓度(20%O2,A组)、低氧浓度Ⅰ组(8%O2,B组)和Ⅱ组(2%O2,C组)及低氧复氧组(2%O2/20%O2,D组)培养的不同时段细胞上清液中VEGF和sFlt-1蛋白的表达。结果:B组、C组于培养72h、96h、120h、144h时间点测得VEGF、sFlt-1的浓度明显高于A组(P<0.01);D组于培养96h、120h、144h后测得VEGF、sFlt-1的浓度较72h低,且分别与A组同期相比无明显差异(P>0.05)。结论:细胞滋养层细胞在低氧条件下VEGF、sFlt-1分泌增加,低氧复氧后VEGF和sFlt-1表达均恢复正常。

Objective： To investigate the expression of vascular endothelial growth factor （VEGF） and soluble fins-like tyrosine kinase receptor 1 （sFlt-1） under different oxygen concentrations in cytotrophoblast cells. Methods： Cytotrophoblast cells of human early-pregnancy were collected by trypsin-collagenase digestion and Percoll gradient centrifugation for primary culture. Cytotrophoblast cells were cultured under group A （normoxic group,20%O2）, group B（8%O2）, group C （2%O2）, group D （hypoxic and reoxygenation group 2%O2/20%O2）, respectively.VEGF and sFlt-1 concentrations at different incubation periods in each culture media were measured using ELISA. Results： The VEGF and sFlt-1 concentrations in group B and group C were significantly higher than those in group A when measured at 72 h, 96 h, 120 h, 144 h in culured （P〈0.01）. VEGF and sFlt-1 concentrations in group D at 96 h, 120 h,144 h in cultured were lower than that at 72 h. The VEGF and sFlt-1 concentrations at 96 h,120 h ,144 h had no obviously difference compared with group A （P〉0.05）. Conclusion： VEGF and sFlt-1 concentrations increase in response to hypoxic, VEGF and sFlt-1 concentratiohs were to recover common after hypoxic and reoxygenation.