丙酮酸钠与贮存红细胞膜磷脂酰丝氨酸暴露的相关性研究
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摘要
目的:探讨丙酮酸钠(SP)与贮存红细胞膜磷脂酰丝氨酸(PS)暴露的相关性。方法:随机选取符合献血标准的健康献血者血液20份,去除白细胞后等量留取2份样本均50 mL/袋(20×2)分为试验组20份(SP组)、对照组20份(NS组)。试验组每袋加入100 mmol/L的SP溶液8.0 mL,对照组加入等量0.9%生理盐水,轻轻混匀制成悬浮红细胞,分别在第1天、第14天、第28天每袋取样1 mL用于流式细胞仪测定红细胞膜表面PS表达;取5 mL采用ELISA法检测上清液中丙二醛(MDA)、三磷酸腺苷(ATP)。结果:红细胞膜表面PS阳性表达率、上清液中MDA都随贮存时间的延长而升高,对照组上升较快,均自第14天开始两者差异有统计学意义(PS:第14天,t=2.763、P<0.05;第28天,t=3.027,P<0.05,MDA:第14天,t=2.763、P<0.05;第28天,t=3.027,P<0.05);上清液中ATP均随时间延长而降低,对照组变化快,自第14天起2组差异有统计学意义(第14天,t=2.385,P<0.05;第28天,t=2.879,P<0.05)。结论:红细胞在贮存过程中添加SP,可提升ATP、减低MDA,能抑制红细胞膜PS暴露,具有改善红细胞贮存损伤的作用。
Objective:To investigate the relationship between sodium pyruvate(SP)and phosphatidylserine exposure of stored erythrocyte membrane. Method:Twenty healthy blood donors who met the standard of blood donation were selected randomly.After leukocyte removal,50 mL/bag(20×2)was used as the experimental group(20×2)and control group(n=20),which was divided into two groups:the experimental group(n=20)and the control group(n=20).SP in each bag of SP solution added with 100 mmol/L,8.0 mL NS added the same amount of 0.9% normal saline,gently mixed into a suspension of red blood cells.1 mL was sampled from each bag and PS expression on erythrocyte membrane surface was measured by flow cytometry at day 1,14 and 28.5 mL supernatant was sampled,and MDA and ATP were determined by ELISA. Result:The positive expression rate of PS on erythrocyte membrane and the MDA in supernatant increased with the prolongation of storage time.The control group rose faster.Since 14 days,the difference between the two groups was statistically significant(PS:14 d t=2.763,P<0.05;28 d t=3.027,P<0.05,MDA:14 d t=2.763,P<0.05;28 d t=3.027,P<0.05).The levels of ATP in the supernatant decreased with the prolongation of the time,and the changes in the control group were fast.Since 14 days,the difference between the two groups was statistically significant(14 d,t=2.385,P<0.05;28 d,t=2.879,P<0.05).Conclusion:The addition of SP to erythrocytes during storage could enhance ATP and decrease MDA,which could inhibit the PS exposure of erythrocyte membrane,and improve the damage of red blood cell storage.
Objective:To investigate the relationship between sodium pyruvate(SP)and phosphatidylserine exposure of stored erythrocyte membrane. Method:Twenty healthy blood donors who met the standard of blood donation were selected randomly.After leukocyte removal,50 mL/bag(20×2)was used as the experimental group(20×2)and control group(n=20),which was divided into two groups:the experimental group(n=20)and the control group(n=20).SP in each bag of SP solution added with 100 mmol/L,8.0 mL NS added the same amount of 0.9% normal saline,gently mixed into a suspension of red blood cells.1 mL was sampled from each bag and PS expression on erythrocyte membrane surface was measured by flow cytometry at day 1,14 and 28.5 mL supernatant was sampled,and MDA and ATP were determined by ELISA. Result:The positive expression rate of PS on erythrocyte membrane and the MDA in supernatant increased with the prolongation of storage time.The control group rose faster.Since 14 days,the difference between the two groups was statistically significant(PS:14 d t=2.763,P<0.05;28 d t=3.027,P<0.05,MDA:14 d t=2.763,P<0.05;28 d t=3.027,P<0.05).The levels of ATP in the supernatant decreased with the prolongation of the time,and the changes in the control group were fast.Since 14 days,the difference between the two groups was statistically significant(14 d,t=2.385,P<0.05;28 d,t=2.879,P<0.05).Conclusion:The addition of SP to erythrocytes during storage could enhance ATP and decrease MDA,which could inhibit the PS exposure of erythrocyte membrane,and improve the damage of red blood cell storage.
引文
[1] 李晓英,芶大明.红细胞衰亡与红细胞膜磷脂酰丝氨酸外翻机制研究进展[J].国际麻醉学与复苏杂志,2016,37(5):464-467.
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[7] Kwiecien S,Konturek PC,Sliwowski Z,et al.Interaction between selective cyclooxygenase inhibitors and capsaicin-sensitive afferent sensory nerves in pathogenesis of stress-induced gastric lesions.Role of oxidative stress[J].J Physiol Pharmacol,2012,63:143-151.
[8] Calderán-Salinas JV,Mufioz-Reyes EG,Guerrero-Romero JF,et al.Eryptosis and oxidative damage in type 2 diabetic mellitus patients with chronic kidney disease[J].Mol Cell Biochem,2011,357:171-179.
[9] 李修良,董鹏,张晔,等.丙酮酸钠盐水对回收自体血液红细胞功能的影响[J].国际麻醉学与复苏杂志,2018,39(1):23-65.
[2] Stowell SR,Smith NH,Zimring JC,et al.Addition of ascorbicacid solution to stored murine red blood cells increases posttransfusion recovery and decreases microparticles and alloimmunization[J].Transfusion,2013,53:2248-2257.
[3] 左珊珊,芶大明,周方强,等.丙酮酸盐对组织细胞的保护作用[J].国际麻醉学与复苏杂志,2014,35(8):757-760.
[4] Beutler E.Energy metabolism and maintenance of erythrocytes.Patrick G G,Bernard GF.The red Cell membrane.Beulter E,Masouredis SP.Preservation and clinical use of erythrocytes and Whole blood.In:Beutler E,Lichtman MA,Coller BS,et al.Williams Hematology[M].6th ed.New York:Mc Graw Hill Co Inc,2001,319-332,333-343,1879-1892.
[5] 牛海玥,王化泉,邵宗鸿,等.红细胞输注研究进展[J].中华医学杂志,2018,98(14):1113-1116.
[6] Cheepala SB,Pitre A,Fukuda Y,et al.The ABCC4 membrane transporter modulates platelet aggregation[J].Blood,2015,126:2307-2319.
[7] Kwiecien S,Konturek PC,Sliwowski Z,et al.Interaction between selective cyclooxygenase inhibitors and capsaicin-sensitive afferent sensory nerves in pathogenesis of stress-induced gastric lesions.Role of oxidative stress[J].J Physiol Pharmacol,2012,63:143-151.
[8] Calderán-Salinas JV,Mufioz-Reyes EG,Guerrero-Romero JF,et al.Eryptosis and oxidative damage in type 2 diabetic mellitus patients with chronic kidney disease[J].Mol Cell Biochem,2011,357:171-179.
[9] 李修良,董鹏,张晔,等.丙酮酸钠盐水对回收自体血液红细胞功能的影响[J].国际麻醉学与复苏杂志,2018,39(1):23-65.