溶血性贫血突变小鼠中锚蛋白外翻介导的红细胞吞噬作用  

Ankyrin Externalization and RGD-α_(v)β_(3) Integrin-Mediated Erythrophagocytosis in Hemolytic Anemic Mutant Mice

作　　者：王琨 韩强 彭书华 彭溦雁 韩林妍 WANG Kun;HAN Qiang;PENG Shuhua;PENG Weiyan;HAN Linyan(Yunyang County People's Hospital,Chongqing,China 404500;The First Affiliated Hospital of Chongqing Medical University,Chongqing,China 400016)

机构地区：[1]重庆市云阳县人民医院,重庆404500 [2]重庆医科大学附属第一医院,重庆400016

出　　处：《中国药业》2022年第10期45-50,共6页China Pharmaceuticals

基　　金：国家自然科学基金[81302307]。

摘　　要：目的探讨红细胞膜蛋白在红细胞程序性死亡过程中的作用。方法选取C57BL/6J野生型(+/+)、基因敲除小鼠锚蛋白基因敲除纯合子型(nb/nb)、带4.2蛋白基因敲除(P4.2-/-)和带3蛋白基因敲除(带3-/-)的小鼠各3对,采用红细胞变形仪检测带3蛋白、带4.2蛋白及锚蛋白缺乏红细胞的综合伸长指数以判定细胞程序性死亡前及死亡过程中的变形能力和渗透脆性;采用流式细胞仪检测锚蛋白及磷脂酰丝氨酸(PS)在红细胞凋亡过程中的外翻情况,以及巨噬细胞对红细胞的黏附作用。结果不同红细胞的综合伸长指数从大到小依次为+/+、nb/nb、带3-/-、P4.2-/-,P4.2-/-红细胞渗透脆性在正常情况及钙离子孵育情况下均较弱;与+/+红细胞比较,P4.2-/-红细胞在生理钙浓度及外加剪切力作用下外翻减弱,nb/nb红细胞中截短锚蛋白的外翻增加;nb/nb红细胞PS外翻明显增加;红细胞黏附数nb/nb为+/+的4~6倍,P4.2-/-低于nb/nb,但高于带3-/-。结论锚蛋白外翻的红细胞更易被巨噬细胞识别,其外翻的精氨酸、甘氨酸和天门冬氨酸基因序列(RGD序列)可能是红细胞程序性死亡的潜在死亡信号。Objective To investigate the role of erythrocyte membrane protein in the process of programmed death of erythrocytes.Methods Mice with C57BL/6J wild-type(+/+),ankyrin gene knockout homozygous(nb/nb),band 4.2 protein gene knockout(P4.2-/-)and band 3 protein gene knockout(band 3-/-)were selected,three pairs of each type.The comprehensive elongation indexes of erythrocytes with band 3 protein,band 4.2 protein and ankyrin lacking erythrocytes were detected by red cell deformer to determine the deformability and osmotic fragility before and during programmed death of cells.The externalization of ankyrin and phosphatidylserine(PS)in the apoptosis of erythrocytes and the adhesion of macrophages to erythrocytes were detected by flow cytometry.Results The comprehensive elongation indexes of different erythrocytes from high to low were as follows:+/+,nb/nb,band 3-/-and P4.2-/-,the osmotic fragility of P4.2-/-erythrocytes was weak under normal conditions and calcium ion incubation.Compared with that of+/+erythrocytes,the externalization of P4.2-/-erythrocytes decreased under the action of physiological calcium concentration and external shear force,and the externalization of truncated ankyrin increased in nb/nb erythrocytes.Compared with that of+/+erythrocytes,the externalization of nb/nb erythrocytes PS increased significantly.The number of erythrocyte adhesion of nb/nb type was 4-6 times that of+/+type,that of P4.2-/-type was lower than that of nb/nb type,but higher than that of band 3-/-type.Conclusion Ankyrin externalization and RGD-α_(v)β_(3) integrin-mediated erythrocytes are more easily recognized by macrophages.The externalization gene sequences of arginine,glycine and aspartate(RGD sequence)may be the potential death signal of the programmed death of erythrocytes.

关 键 词：RGD序列 α_(V)β_(3)整合素 锚蛋白 带3蛋白 带4.2蛋白 基因敲除 

分 类 号：R96[医药卫生—药理学] R556.6[医药卫生—药学]