氢气保存的红细胞悬液对失血性休克大鼠的复苏效果

Hydrogen-preserved red blood cell suspension improves resuscitation outcomes in rats with hemorrhagic shock

摘要

目的 探讨氢气对大鼠红细胞的保护作用及其对失血性休克复苏的效果。方法 将大鼠红细胞分为常规保存组(n=6)和氢气保存组(n=6), 分别于保存第1、4、7、11、14日测定24 h输血后回收率(24 h-PTR); 在光镜和电镜下观察红细胞形态; 检测体外保存红细胞的质量指标。另取失血性休克大鼠, 分为常规复苏组(n=6)和氢气复苏组(n=6), 分别输注常规保存和氢气保存的红细胞进行复苏。于静息状态、休克末及复苏后监测生命体征, 检测肾损伤和炎症因子水平, 统计输血量。结果 氢气保存组红细胞的24 h-PTR从保存第7日起高于常规保存组(P<0.05)。光镜下, 氢气保存组异形红细胞及细胞碎片少于常规保存组; 电镜下, 氢气保存组红细胞膜表面超微结构损伤明显减轻。氢气保存组红细胞的游离血红蛋白和溶血率分别从第4、7日起低于常规保存组; 三磷酸腺苷、2,3二磷酸甘油酸含量分别从第7、14日起高于常规保存组; 超氧化物歧化酶、谷胱甘肽过氧化物酶活性分别从第4、11日起高于常规保存组; 还原型谷胱甘肽和过氧化氢酶活性从第14日起高于常规保存组(P<0.05)。复苏后2 h, 氢气复苏组大鼠的平均动脉压高于常规复苏组, 心率和输血量均低于常规复苏组(P<0.05)。复苏后24 h, 氢气复苏组的血肌酐、血尿素氮及多数炎症因子水平均低于常规复苏组(P<0.05), 仅干扰素γ差异无统计学意义。肾脏HE染色显示, 氢气复苏组基本正常, 常规复苏组可见肾小管上皮细胞坏死、脱落及管型形成。结论 氢气可提升红细胞体外保存质量, 输注氢气保存的红细胞悬液能够实现更优的失血性休克复苏效果。

Abstract

Objective To investigate the protective effect of hydrogen on rat red blood cells (RBCs) and its efficacy in hemorrhagic shock resuscitation. Methods Rat RBCs were divided into conventional storage group (n=6) and hydrogen storage group (n=6). The 24-hour post-transfusion recovery (24 h-PTR) was measured on days 1, 4, 7, 11, and 14 of storage. RBC morphology was observed under light microscopy and electron microscopy. Quality indicators of stored RBCs were assessed in vitro. Separately, rats with hemorrhagic shock were divided into conventional resuscitation group (n=6) and hydrogen resuscitation group (n=6), receiving resuscitation with conventionally stored RBCs or hydrogen-stored RBCs, respectively. Vital signs were monitored at rest, at the end of shock, and after resuscitation. Renal injury and inflammatory cytokine levels were measured, and the blood transfusion volume was recorded. Results The 24 h-PTR of RBCs in the hydrogen storage group was higher than that in the conventional storage group from day 7 of storage onward (P<0.05). Under light microscopy, the hydrogen storage group showed fewer deformed RBCs and cell fragments than the conventional storage group; under electron microscopy, ultrastructural damage to the RBC membrane surface was markedly reduced in the hydrogen storage group. Free hemoglobin and hemolysis rate in the hydrogen storage group were lower than those in the conventional storage group from day 4 and day 7, respectively. Adenosine triphosphate and 2,3 diphosphoglycerate levels were higher in the hydrogen storage group from day 7 and day 14, respectively. Superoxide dismutase and glutathione peroxidase activities were higher from day 4 and day 11, respectively, and reduced glutathione and catalase activities were higher from day 14 in the hydrogen storage group (P<0.05). At 2 h after resuscitation, mean arterial pressure in the hydrogen resuscitation group was higher, while heart rate and blood transfusion volume were lower than those in the conventional resuscitation group (P<0.05). At 24 h after resuscitation, serum creatinine, blood urea nitrogen, and most inflammatory cytokine levels were lower in the hydrogen resuscitation group than those in the conventional resuscitation group (P<0.05), except for interferon γ, which showed no statistically significant difference. Kidney HE staining revealed nearly normal morphology in the hydrogen resuscitation group, whereas the conventional resuscitation group exhibited necrosis, shedding of renal tubular epithelial cells, and cast formation. Conclusion Hydrogen improves the quality of RBCs during storage in vitro. Infusion of hydrogen-stored RBC suspension achieves better outcomes in hemorrhagic shock resuscitation.

关键词

氢气 / 红细胞悬液 / 失血性休克 / 复苏术 / 氧化性应激 / 冷温贮藏 / 疾病模型, 动物 / 治疗结果

Key words

hydrogen / erythrocyte suspension / hemorrhagic shock / resuscitation / oxidative stress / cold storage / disease models, animal / treatment outcome

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基金资助

国家卫生健康委医药卫生科技发展研究项目(WKZX2024CX301202)