PNH患者红细胞膜乙酰胆碱酯酶缺失的研究

ENGLISH ABSTRACTS OF ORIGINAL ARTICLESDEFICIENCY OF ACETYLCHOLINESTERASE INERYTHROCYTE MEMBRANE OF PAROXYSMAL NOCTURNAL HEMOGOBINURIA PATIENTS

用间接免疫荧光流式细胞术测定阵发性睡眠性血红蛋白尿症（ＰＮＨ）患者和正常人红细胞乙酰胆碱酯酶（ＡｃｈＥ）表达量，发现ＰＮＨ的ＡｃｈＥ表达远远低于正常，而且蛇毒溶血试验与ＡｃｈＥ的缺失呈正相关。由于ＰＮＨ正常和异常血细胞同时并存，为进一步研究ＰＮＨ的ＡｃｈＥ（－）红细胞，采用ＰｒｏｔｅｉｎＡＳｅｐｈａｒｏｓｅ６ＭＢ结合ＡｃｈＥ单抗亲和层析法分离出ＰＮＨ的ＡｃｈＥ（－）红细胞。用￣３Ｈ－肌醇标记实验证明，正常红细胞膜在区带４．１有较高放射活性，而ＰＮＨ红细胞膜放射活性极低，ＰＮＨ的ＡｃｈＥ（－）红细胞膜完全无放射活性；用免疫印渍实验证明区带４．１是ＡｃｈＥ存在部位。用酶联免疫方法直接检测ＰＮＨ患者和正常人血浆中ＡｃｈＥ含量，结果发现ＰＮＨ患者血浆中有ＡｃｈＥ存在，而正常人未能检出。

aroxysmal nocturnal hemoglobinuria(PNH ) is anacquired clonal hematologic disorder,In order to clari-fy the mechanism of the hemolysis of PNH erythro-cytes,acetylcholinesterase(AchE)expression on PNHand normal erythrocyte membrane was analysed byflowcytometry. It was found that the AchE expressionon PNH ervthrocytes was muchlower tban normal.Hemolysis induced by Cobra venom factor(Cof) is pro-portional to AchE deficiency. Anti-AchE monocionalantibody bound affinity chromatography(Protein ASepharose 6MB ) used to separate AchE(-)from otherPNH erythrocytes. Erythrocyte membrane labelledwith ￣H-inositol showed that radioactivity was at band4.1 in normal erythrocytes , but very low at the sameband in PNH erythrocytes and totally undetectable inPNH AcbE(一)erythrocytes,By ELISA method as-say,AchE was detected in plasma of PNH patients , butundetected in plasma of normal persons.