METHODS: Guinea pig RBCs were collected from whole blood (WB) treated with varying UV doses (10, 20, 40 or 80 J/mL) in the presence of riboflavin (UVR-RBCs). In vitro tests for UVR-RBCs included hemolysis, osmotic fragility, and cellular morphology by scanning electron microscopy. Guinea pigs transfused with one-day post-treatment UVR-RBCs were evaluated for plasma hemoglobin (Hb), non-transferrin bound iron (NTBI), total iron and Perls-detectable hemosiderin deposition in the spleen and kidney, and renal uptake of Hb.
RESULTS: Acute RBC injury was dose dependently accelerated after treatment with UV light in the presence of riboflavin. Aberrant RBC morphology was evident at 20, 40, and 80 J/mL, and membrane lysis with Hb release was prominent at 80 J/mL. Guinea pigs transfused with 40 and 80 J/mL UVR-RBCs showed increased plasma Hb levels, and plasma NTBI was elevated in all UVR-RBC groups (10-80 J/mL). Total iron levels and Perls-hemosiderin staining in spleen and kidney as well as Hb uptake in renal proximal tubules were increased 8 hours post-transfusion with 40 and 80 J/mL UVR-RBCs.
CONCLUSIONS: UVR-RBCs administered to guinea pigs increased markers of intravascular and extravascular hemolysis in a UV dose-dependent manner. This model may allow for the discrimination of RBC injury during testing of extensively processed RBCs intended for transfusion.
方法:在核黄素(UVR-RBC)的存在下,从用不同UV剂量(10、20、40或80J/mL)处理的全血(WB)收集豚鼠RBC。UVR-RBC的体外测试包括溶血,渗透脆弱性,扫描电镜和细胞形态。用治疗后一天的UVR-RBC输注的豚鼠评估血浆血红蛋白(Hb),非转铁蛋白结合铁(NTBI),总铁和Perls-可检测的含铁血黄素在脾脏和肾脏中的沉积,和肾吸收Hb。
结果:在核黄素存在下用紫外光治疗后,急性红细胞损伤呈剂量依赖性加速。在20、40和80J/mL时,红细胞形态异常明显,Hb释放的膜溶解在80J/mL时显著。输入40和80J/mLUVR-RBC的豚鼠显示血浆Hb水平升高,所有UVR-RBC组(10-80J/mL)的血浆NTBI均升高。在输注40和80J/mLUVR-RBC后8小时,脾脏和肾脏中的总铁水平和Perls-含铁血黄素染色以及肾脏近端小管中的Hb摄取增加。
结论:给予豚鼠UVR-RBC以紫外线剂量依赖性方式增加血管内和血管外溶血的标志物。该模型可以允许在用于输血的广泛处理的RBC的测试期间区分RBC损伤。