电镜分析非晶状体βγ晶状体蛋白与三叶因子蛋白复合物在人红细胞膜上的孔道形成效应  

Electron Microscopic Analysis of βγ-CAT Pore-forming Effect on Human Erythrocyte Membrane

作　　者：刘树柏[1,2] 何英英[1,2] 钱金桥[1,2,3] 张云[1] 

机构地区：[1]中国科学院昆明动物研究所动物模型与人类疾病机理重点实验室生物毒素研究室,云南昆明650223 [2]中国科学院研究生院,北京100049 [3]昆明医学院第一附属医院麻醉科,云南昆明650032

出　　处：《Zoological Research》2008年第5期503-510,共8页动物学研究（英文）

基　　金：国家自然基金(30630014;30570359);中国科学院重要研究方向基金(KSCX2-YW-R-088)

摘　　要：非晶状体βγ晶状体蛋白与三叶因子蛋白复合物(non-lens βγ-crystallin and trefoil factor complex,βγ-CAT)是一个从大蹼铃蟾(Bombina maxima)皮肤分泌物中分离的天然分子量为72kDa的全新的蛋白复合物。本研究测定了βγ-CAT处理红细胞后引起细胞内钾离子外流与溶血效应的时效曲线,结合扫描电子显微镜和透射电子显微镜分析βγ-CAT处理红细胞引起的早期形态学变化。结果表明:βγ-CAT(3nmol/L)37℃处理红细胞5min,(93.31±5.89)%的细胞内钾离子迅速外流(P<0.01),相应溶解率为(13.12±1.92)%(P<0.05)。电子显微镜观察发现红细胞形态发生明显变化,细胞体积增加,肿胀。少数红细胞表面向外形成棘状异常突起,部分肿胀细胞内的血红蛋白通过棘状突起缺口向细胞外喷射血红蛋白。表明βγ-CAT通过在红细胞膜上形成孔道使细胞内钾离子迅速外流导致红细胞内渗透压改变而溶血。其结果为理解βγ-CAT的溶血机制提供了直接的形态学证据。βγ-CAT is a naturally existing multifunctional protein complex of non-lens βγ-crystallin and trefoil factor from Bornbina maxima skin secretions. In this paper, we first analysed the time-response curve of hemolysis and intracellular potassium effiux induced by βγ-CAT on human erythrocytes. After βγ-CAT （3 nmol/L） in human erythrocytes was treated at 37℃ for 5rain, we found that about 93.31 ±5.89% （P〈0.01） of the intracellular potassium ions leaked, whereas the percentage of hemolysis was onlyl 3.12± 1.92 % （P〈0.05）. We employed electron microscopy to observe the erythrocytes＇ morphological changes and found erythrocytes were swollen and some had globular structures occurring on the cell surfaces, and fast releasing hemoglobins. These results indicated that the hemolytic effect of βγ-CAT on human erythrocyte membranes was due to intracellular potassium ion＇s rapid efflux. These results demonstrate morphological proof in understanding the mechanism of βγ-CAT pore-forming effect on human erythrocytes.

关 键 词：三叶因子 非晶状体βγ-晶状体蛋白 非品状体βγ-晶状体蛋白与三叶因子蛋白复合物 孔道形成蛋白 扫描电子显微镜 投射电子显微镜 

分 类 号：R776[医药卫生—眼科]