Mouse erythrocytes as carriers for coencapsulated alcohol and aldehyde dehydrogenase obtained by electroporation -: In vivo survival rate in circulation, organ distribution and ethanol degradation
Mouse erythrocytes as carriers for coencapsulated alcohol and aldehyde dehydrogenase obtained by electroporation -: In vivo survival rate in circulation, organ distribution and ethanol degradation
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DOI：
10.1016/s0024-3205(01)00991-2
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发表时间：
2001-03-16
期刊：
影响因子：
6.1
通讯作者：
Pinilla, M
中科院分区：
文献类型：
--
作者：
Lizano, C;Pérez, MT;Pinilla, M
用电穿孔技术将乙醇脱氢酶和乙醛脱氢酶（ADH和ALDH）共囊化到小鼠红细胞中。获得的最佳条件如下：420 V，每15分钟4次1 ms脉冲，37 ℃，通过重新密封完成：37 ℃，1 h。对于ADH + ALDH负载的红细胞，获得范围为11-12%的包封率。载体细胞回收率为52%。扫描电镜下观察到的电穿孔红细胞表现出内陷球形口细胞的趋势。在电穿孔/重新密封的RBC中未发现这些内陷。Cr-51-RBC的静脉内施用表现出双峰药代动力学特征：初始阶段（t（1/2 α）），血浆Cr-51-RBC快速下降，随后是缓慢和延长的消除期（t（1/2 beta）），消除期的体内存活率值表明，电穿孔Cr-51负载的RBC的存活率略低（t（1/2 β），4.5天）。电穿孔Cr-51-RBC（未加载和加载）血液的平均清除率值（分别为0.51%Cr-51/天和0.54%Cr-51/天）高于天然Cr-51-RBC的平均清除率值（0.18%Cr-51/天）。证明电穿孔RBC的靶器官与天然RBC相同。然而，电穿孔的RBC在肝、脾和肺中显示出最高的积累，因为它们被网状内皮系统迅速识别。诱导至急性乙醇中毒状态并用ADH + ALDH-RBC处理的小鼠清楚地显示出血浆中乙醇浓度水平（小于43%乙醇）低于用天然RBC处理的中毒小鼠。因此，用ADH + ALDH-RBC处理的中毒小鼠血液中乙醇的清除值（0.39 ml/min）高于用天然RBC处理的小鼠（0.20 ml/min）。所获得的结果表明，ADH + ALDH负载的红细胞可以被用作一个潜在的载体系统，在体内去除高水平的乙醇从血液中引起的过量饮酒。(C)2001 Elsevier Science Inc. All rights reserved.
Alcohol dehydrogenase and aldehyde dehydrogenase (ADH and ALDH) have been coencapsulated into mouse erythrocytes by an electroporation technique. The optimal conditions were achieved as follows: 420 V, four pulses of 1 ms every 15 min. at 37 degreesC, completed by resealing: 1 h at 37 degreesC. An encapsulation yield ranging from 11-12% was obtained for ADH + ALDH-loaded erythrocytes. Carrier cell recovery was 52%. Electroporated-RBCs observed under Scanning electron microscopy exhibited a tendency toward invaginated sphero-stomatocytes. These invaginations were not found in electroporated/resealed RBCs, The intravenous administration of Cr-51-RBCs manifested a bimodal pharmacokinetic profile: an initial phase (t(1/2 alpha)) with a rapid decrease of plasma Cr-51-RBCs followed by a slow and prolonged elimination phase (t(1/2 beta)), The values corresponding to in vivo survival rate during the elimination phase indicated that the survival rate of Cr-51-electroporated loaded-RBCs was slightly lower (t(1/2 beta), 4.5 days) than Cr-51-native RBCs (t(1/2 beta), 5.3 days). The mean clearance values from blood of electroporated Cr-51-RBCs (unloaded and loaded) were higher (0.51 %Cr-51/day and 0.54 %Cr-51/day, respectively) than the obtained for native Cr-51-RBCs (0.18 % Cr-51/day). The target organs for electroporated RBCs proved to be the same as for native RBCs. However, electroporated RBCs showed highest accumulation in liver, spleen and lung, since they were promptly recognized by the reticuloendothelium system. Mice induced to the state of acute ethanol intoxication and treated with ADH + ALDH-RBCs clearly showed a lower level of ethanol concentration in plasma (less than 43% ethanol) than the intoxicated mice treated with native RBCs. En consequence the clearance values of ethanol from blood in intoxicated mice treated with ADH + ALDH-RBCs (0.39 ml/min) were higher than the treated with native RBCs (0.20 ml/min). The results obtained suggest that ADH + ALDH-loaded erythrocytes could be used as a potential carrier system for in vivo removal of high levels of ethanol from blood caused by excessive alcohol consumption. (C) 2001 Elsevier Science inc. All rights reserved.