【摘要】 利用γ-32P-ATP5、1Cr示踪法研究了蛋白激酶C(Protein Kinase C,PKC)对血管内皮细胞CD44分子表达及内皮粘附功能的影响及机制。通过测定外源性底物对γ3-2P-ATP的摄入量确定人脐静脉内皮细胞(Human Umbilical Vein Endothelial Cells,HUVECs)PKC的活性;以γ-32P-ATP及聚丙烯酰胺凝胶电泳测定CD44分子的磷酸化;用流式细胞仪测定CD44分子的表达;以51Cr标记血小板测定HUVECs的粘附率。结果表明,用PKC激活剂十四酰佛波醇乙酯(Phorbol-myristate-acetate,PMA)作用人HUVECs 30 min,PKC的活性达到最高,CD44的磷酸化达最大,为4 960±136 min-1.μg-1,CD44分子表达及HUVECs的粘附率也达到最高水平;与对照组相比,P均<0.001。这说明PKC能通过磷酸化作用上调CD44分子的表达,并增强内皮细胞的粘附功能。更多还原

【Abstract】 In order to study the effects of protein kinase C（PKC） on CD44 expression in vascular endothelial cells and the consequent effects on endothelial adhesion,human umbilical vein endothelial cells（HUVECs） are taken for study model.PKC activity is determined by the incorporation of γ-³²P-ATP into exogenous substrate.Phosphorylation of CD44 is detected by the incorporation of ³²P into CD44.CD44 molecules expression is analysed by flow cytometry.Platelets marked by ⁵¹Cr measurated HUVECs adhesive ability.The results show that phosphorylation of CD44 achieved the highest level after PKC is activated for 30 minutes,CD44 molecules expression and adhesive percentage of HUVECs to platelets also reached the higheast value simultaneously.It suggests that PKC upregulates CD44 molecules expression by phosphorylation,and raises the adhesive ability of vascular endoyhelial cells.更多还原