膜联蛋白A5对t-PA联合纤溶酶原诱导血-脑屏障通透性的抑制作用  

Inhibitory effect of annexin A5 on permeability of blood-brain barrier induced by t-PA and plasminogen

作　　者：张锐斌 王鹏 李飞[1] 张般若 ZHANG Ruibin;WANG Peng;LI Fei(College of Pharmacy,Nanjing Medical University,Nanjing 211166,CHINA)

机构地区：[1]南京医科大学药学院,江苏211166 [2]江苏先声药业有限公司

出　　处：《江苏医药》2023年第9期865-868,873,F0002,共6页Jiangsu Medical Journal

基　　金：国家自然科学基金(82171224)。

摘　　要：目的探讨膜联蛋白A5对组织型纤溶酶原激活剂(t-PA)联合纤溶酶原诱导血-脑屏障通透性的抑制作用及其可能机制。方法在Transwell细胞小室上培养人脑微血管内皮细胞株HCMEC/D3,加入不同浓度t-PA 80、160和320 nmol/L联合纤溶酶原50 nmol/L作用6、12和24 h,同时设对照组,观察细胞模型通透性和细胞活力。随后再加入不同浓度膜联蛋白A550、100、200和500 nmol/L,观察细胞模型通透性的变化,并采用Western blot法检测紧密连接蛋白1(ZO-1)的蛋白表达。结果与对照组相比,t-PA 160 nmol/L+纤溶酶原50 nmol/L组和t-PA 320 nmol/L+纤溶酶原50 nmol/L组细胞模型通透性均呈时间依赖性的增加(P<0.05或P<0.01),并且作用24 h后细胞活力降低(P<0.01)。与t-PA 160 nmol/L+纤溶酶原50 nmol/L组相比,t-PA 160 nmol/L+纤溶酶原50 nmol/L+膜联蛋白A5100 nmol/L组和t-PA 160 nmol/L+纤溶酶原50 nmol/L+膜联蛋白A5200 nmol/L组细胞模型通透性降低(P<0.05或P<0.01)。t-PA 160 nmol/L+纤溶酶原50 nmol/L组ZO-1蛋白表达低于对照组(P<0.01);而t-PA 160 nmol/L+纤溶酶原50 nmol/L+膜联蛋白A5100 nmol/L组ZO-1蛋白表达高于t-PA 160 nmol/L+纤溶酶原50 nmol/L组(P<0.01)。结论膜联蛋白A5能够抑制t-PA联合纤溶酶原诱导血-脑屏障通透性的增加;其机制可能与上调内皮细胞间ZO-1的表达有关。Objective To investigate the inhibitory effect and underlying mechanism of annexin A5 on the permeability of blood-brain barrier induced by tissue plasminogen activator(t-PA)and plasminogen.Methods Human brain microvascular endothelial cell line HCMEC/D3 was cultured in Transwell chamber,which was then stimulated with different concentrations of t-PA 80,160,320 nmol/L and plasminogen 50 nmol/L for 6,12 and 24 hours.A control group was set as well.The intercellular permeability and cell viability were observed.Then the different concentrations of annexin A550,100,200 and 500 nmol/L were added for observing the change of intercellular permeability.The protein expression of ZO-1 was detected by Western blot.Results Compared with control group,the intercellular permeability was increased in a time-dependent manner(P<0.05 or P<0.01),and the cell viability was decreased after 24 hours in t-PA 160 nmol/L+plasminogen 50 nmol/L group and t-PA 320 nmol/L+plasminogen 50 nmol/L group(P<0.01).Compared with t-PA 160 nmol/L+plasminogen 50 nmol/L group,the intercellular permeability was decreased in t-PA 160 nmol/L+plasminogen 50 nmol/L+annexin A5100 nmol/L group and t-PA 160 nmol/L+plasminogen 50 nmol/L+annexin A5200 nmol/L group(P<0.05 or P<0.01).The protein expression of ZO-1 in t-PA 160 nmol/L+plasminogen 50 nmol/L group was lower than that in control group(P<0.01),while which in t-PA 160 nmol/L+plasminogen 50 nmol/L+annexin A5100 nmol/L group was higher than that in t-PA 160 nmol/L+plasminogen 50 nmol/L group(P<0.01).Conclusion Annexin A5 can inhibit the increase of the permeability of blood-brain barrier induced by t-PA and plasminogen,which may be related with the upregulated expression of ZO-1 between endothelial cells.

关 键 词：膜联蛋白A5 组织型纤溶酶原激活剂 血-脑屏障 

分 类 号：R743[医药卫生—神经病学与精神病学]