【摘要】 目的研究低氧复氧诱导培养的人脐静脉内皮细胞(HUVECs)表达趋化因子FKN与Wnt信号通路中关键信号分子β-catenin、GSK-3β的相关性。方法 HUVECs,随机分为对照组、低氧复氧组和氯化锂(LiCl,10 mmol/L)组。免疫细胞荧光FITC法检测FKN蛋白表达,免疫细胞化学技术检测GSK-3β及β-catenin的表达。RT-PCR技术检测GSK-3β和FKN mRNA表达。结果与正常对照组比较,低氧复氧引起GSK-3β、β-catenin及FKN表达明显增加(P<0.05),于复氧2 h达表达高峰;与低氧复氧组比较,LiCl组β-catenin表达明显增加(P<0.05),FKN与GSK-3β表达明显降低(P<0.05),但仍然高于正常对照组。LiCl组FKN mRNA表达高于正常对照组(P<0.05)而低于低氧复氧组(P<0.05),于复氧1 h达表达高峰;GSK-3βmRNA在低氧前后及复氧各时间点无差异表达。结论 Wnt信号通路的激活可能在转录后翻译阶段参与LiCl下调低氧诱导HUVECs表达FKN的过程。更多还原

【Abstract】 Objective To investigate the relationship between Wnt signaling pathway and hypoxia-induced fractalkine expression in human umbilical vein endothelial cells(HUVECs). Methods HUVECs were cultivated and divided into three groups i. e. randomly( control group,hypoxia / reoxygenation group,LiCl 10 mmol / L group),which were used to establish the hypoxia / reoxygenation models. The mRNA expressions of fractalkine and GSK-3βwere assessed by RT-PCR. The intracellular distribution and expression of fractalkine were determined by immunofluorescent staining. The expression of GSK-3β and β-catenin were analyzed with immunocytochemistry. Results In hypoxia / reoxygenation group,the expression of GSK-3β and β-catenin were increased(P < 0. 05),the mRNA and protein expression of fractalkine increased remarkably(P < 0. 05). In LiCl groups,the mRNA and protein expression of fractalkine reduced significantly(P < 0. 05),the expression of GSK-3β reduced but β-catenin increased relativitily(P < 0. 05). However,the mRNA expression of GSK-3β had no statistically difference in all groups.Conclusions The hypoxia damages HUVECs through upregulating fractalkine expression. LiCl pretreatment inhibits fractalkine expression proportional. Its mechanism may be related to the activation of Wnt signaling pathway.更多还原