摘要
目的 本研究旨在探讨Sema7a蛋白促进内皮细胞和巨核细胞之间粘附的分子机制。方法 通过使用人脐静脉内皮细胞(human umbilical vascular endothelial cells,HUVECs)和巨核细胞系MEG01在体外模拟肺部血管和巨核细胞的粘附,使用免疫荧光、流式细胞术、4D非标记定量蛋白质组学分析等技术,检测内皮细胞与巨核细胞之间的粘附、HUVECs表面的Sema7a蛋白结合情况,及与Sema7a蛋白结合后的HUVECs蛋白质表达差异及相关生物学信息的变化,预测潜在的信号通路。并利用免疫荧光技术检测HUVECs的细胞间粘附分子1(intercellular cell adhesion molecule-1,ICAM-1)和血管黏附分子1(vascular cell adhesion molecule-1,VCAM-1)的表达。结果 Sema7a蛋白与HUVECs结合后,激活了HUVECs细胞MAPK信号通路,并上调了HVUECs的黏附分子ICAM-1和VCAM-1的表达,促进了巨核细胞MEG01与HUVECs的粘附。结论 Sema7a蛋白通过上调内皮细胞黏附分子ICAM-1和VCMA-1的表达,促进了巨核细胞MEG01与内皮细胞HUVECs间的粘附。
Objective The purpose of this study was to explore the mechanism of Sema7a protein promoting the adhesion between endothelial cells and megakaryocytes.Methods Human umbilical vascular endothelial cells(HUVECs)and megakaryocyte line MEG01 were used to simulate the adhesion of pulmonary vessels and megakaryocytes in vitro.Immunofluorescence,flow cytometry,4D Label free and other biological techniques were used to detect the adhesion between endothelial cells and megakaryocytes,the amount of Sema7a binding to HUVECs,and the changes of protein expression and biological information in HUVECs.Immunofluorescence was used to detect the expression of intercellular cell adhesion molecule-1(ICAM-1)and vascular cell adhesion molecule-1(VCAM-1)in HUVECs.Results The adhesion between MEG01 and HUVECs was promoted after Sema7a binding to HUVECs,with the MAPK signal pathway been activated,and molecules ICAM-1 and VCMA-1 of HUVECs been up-regulated.Conclusion Sema7a promoted the adhesion between megakaryocytes MEG01 and endothelial cells HUVECs by up-regulating the expression of molecules ICAM-1 and VCMA-1 in HUVECs.
作者
赖冬娣
董涵
魏亚明
苑召虎
LAI Dongdi;DONG Han;WEI Yaming;YUAN Zhaohu(Department of Blood Transfusion,the Second Affiliated Hospital,School of Medicine,South China University of Technology,Guangzhou,Guangdong 510180;Department of Blood Transfusion,Guangzhou First Peoples Hospital,Guangzhou,Guandong 510180;Guangdong Engineering Research Centre of Precise Transfusion,Guangzhou,510180)
出处
《临床输血与检验》
CAS
2024年第3期299-308,共10页
Journal of Clinical Transfusion and Laboratory Medicine
基金
广东省自然科学基金省企联合项目(No.2022A1515220019、No.2021A1515220044)资助。
