阵发性睡眠性血红蛋白尿症红细胞乙酰胆碱酯酶的研究

Studies on Acetylcholinesterase of Paroxysmal Nocturnal Hemoglobinuria Erythrocyte

用化学方法测定了乙酰胆碱脂酶（ＡｃｈＥ）活性，阵发性睡眠性血红蛋白尿症（ＰＮＨ）红细胞远低于正常红细胞。为了进一步研究ＰＮＨＡｃｈＥ（—）的红细胞，采用Ｐｒｏｔｅｉｎ Ａ Ｓｅｐｈａｒｏｓｅ ６ＭＢ结合ＡｃｈＥ单抗亲和层析法分离出ＰＮＨＡｃｈＥ（—）的红细胞。用间接免疫荧光流式细胞术检测，ＰＮＨ细胞ＡｃｈＥ低于正常，而ＰＮＨＡｃｈＥ（—）红细胞未能检出ＡｃｈＥ。３Ｈ－肌醇标记实验证明，正常红细胞膜区带４．１处有较高的放射活性，而ＰＮＨ红细胞极低，ＰＮＨＡｃｈＥ（—）红细胞完全无放射活性。用ＡｃｈＥ抗体做免疫印渍实验证明了ＡｃｈＥ存在区带４．１部位。ＤＭＰＣ诱导正常和ＰＮＨ红细胞，检测二者囊泡化的程度，发现ＰＮＨ病人红细胞远比正常人红细胞易于囊泡化。

PNH is an acquired hemolytic disorder. In order to understand the mechanism of the hemolysis of PNH erythrocytes. AchE activity on PNH and normal erythrocytes membrane was analysed by chemical method. A device utilizing monoclonal AchE antibody and affinity chromatography (protein A sepharose 6 MB) was used to Separate acetylcholinesterase deficient from PNH total erythrocytes. The Surface AchE on membrane of PNH and normal erythrocytes was analysed by a flowcytometer. It was found that loss of AchE from erythrocytes membrane of PNH. Erythrocyte membrane was labelled with ￣3H-inositol.The distribution and amount of phosphatidylinositol anchored protein in erythrocyte membrane of normal and PNH erythrocyte were determined. Radioactivity was found at band 4. 1 in normal erythrocyte. Radioactivity in PNH erythrocyte membrane was much lowerthan normal and was not detectable in PNH AchE(─). Vesiculation of erythrocyte induced by DMPC was also detected. It was found that PNH erythrocyte was much easier to be vesiculated than normal.