Goodrich R P, Sowemimo-Coker S O, Zerez C R, Tanaka K R
Department of Basic Research, Cryopharm Corporation, Pasadena, CA 91107.
Proc Natl Acad Sci U S A. 1992 Feb 1;89(3):967-71. doi: 10.1073/pnas.89.3.967.
Normal human erythrocytes (RBC) were freeze-dried under conditions that caused minimal modification in normal RBC metabolic activities. Because of the known effects of long-term storage on metabolic activities, we studied the effects of our lyophilization process on RBC metabolism. Of all the metabolic enzymes studied, only triosephosphate isomerase (D-glyceraldehyde-3-phosphate ketol-isomerase, EC 5.3.1.1), enolase (2-phospho-D-glyceratehydro-lyase, EC 4.2.1.11), and pyruvate kinase (ATP:pyruvate O2-phosphotransferase, EC 2.7.1.40) were decreased when compared with fresh control nonlyophilized RBC. The activities of these enzymes did not differ significantly from those of blood bank RBC. Concentrations of high-energy intermediates, ATP, and 2,3-diphosphoglycerate, along with lactate and ATP production were decreased in lyophilized RBC. No enzymes of the pentose phosphate shunt were altered during lyophilization. In addition, our data show that lyophilized RBC possess an intact capacity to (i) synthesize adenine nucleotides and (ii) reduce MetHb to Hb and, thus, maintain the Hb in a functional physiologic state similar to fresh nonlyophilized RBC. The present study demonstrates the possibility of lyophilizing RBC in a manner that maintains normal metabolic and enzymatic function upon rehydration.
正常人类红细胞（RBC）在对正常RBC代谢活动造成最小程度改变的条件下进行冻干。由于长期储存对代谢活动的已知影响，我们研究了冻干过程对RBC代谢的影响。在所有研究的代谢酶中，与新鲜对照未冻干的RBC相比，只有磷酸丙糖异构酶（D-甘油醛-3-磷酸酮醇异构酶，EC 5.3.1.1）、烯醇化酶（2-磷酸-D-甘油酸水解酶，EC 4.2.1.11）和丙酮酸激酶（ATP：丙酮酸O2-磷酸转移酶，EC 2.7.1.40）活性降低。这些酶的活性与血库RBC的活性没有显著差异。冻干RBC中高能中间体、ATP和2,3-二磷酸甘油酸的浓度以及乳酸和ATP的产生均降低。冻干过程中磷酸戊糖途径的酶没有改变。此外，我们的数据表明，冻干RBC具有完整的能力：（i）合成腺嘌呤核苷酸，（ii）将高铁血红蛋白还原为血红蛋白，从而将血红蛋白维持在与新鲜未冻干RBC相似的功能性生理状态。本研究证明了以一种在复水后能维持正常代谢和酶功能的方式冻干RBC的可能性。
