Effects of saline washing and MAP resuspension on protein expression of erythrocyte membrane
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摘要: 目的 研究运用甘露醇-腺嘌呤-磷酸盐红细胞保养液(mannitol adenine phosphate red blood cell maintenance solution，MAP)重悬去白细胞洗涤红细胞对红细胞pH值、膜蛋白CD47分子和磷脂酰丝氨酸(phosphatidylserine，PS)表达变化。方法 随机选取10袋全血，滤除白细胞后，每袋去白细胞全血平均分成2份，分别制备成去白细胞悬浮红细胞和MAP去白细胞洗涤红细胞，分别于采血第2、7、14、21、28和35天，检测CD47蛋白平均荧光强度、PS蛋白阳性率和pH值。结果 去白细胞洗涤红细胞组CD47平均荧光强度降低，第2、7、14和21天与去白细胞悬浮红细胞第35天比较，差异有统计学意义(457.35±81.67、421.93±54.01、304.03±26.08、288.79±107.07 vs 209.81±26.17，P < 0.05)；去白细胞洗涤红细胞组PS蛋白阳性率逐渐升高，第2、7、14、21和35天与去白细胞悬浮红细胞第35天比较，差异有统计学意义(0.08±0.13、0.07±0.13、0.03±0.02、1.34±0.56、4.14±1.53 vs 2.29±0.31，P < 0.05)；相同时间点MAP去白洗涤红细胞pH值均低于去白细胞悬浮红细胞，差异有统计学意义(第2天：6.82±0.04 vs 6.9±0.09、第7天：6.64±0.04 vs 6.72±0.07、第14天：6.57±0.03 vs 6.66±0.08、第21天：6.49±0.02 vs 6.58±0.04、第35天：6.45±0.02 vs 6.52±0.03，均P < 0.05；第28天：6.46±0.02 vs 6.55±0.03，P < 0.01)。结论 保存期内MAP去白细胞洗涤红细胞膜CD47蛋白表达降低，PS蛋白外翻增加，损伤加重与pH相关，建议第21天为MAP去白洗涤红细胞最佳保存期。
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关键词:
- 甘露醇-腺嘌呤-磷酸盐红细胞保养液 /
- 洗涤红细胞 /
- CD47 /
- 磷脂酰丝氨酸
Abstract: Objective To investigate the effects of saline washing and(mannitol adenine phosphate red blood cell maintenance solution, MAP)resuspension red blood cells on pH value and expression of membrane protein CD47 and phosphatidyl serine(PS).Methods Ten bags of whole blood were randomly selected, each bag of whole blood was prepared into leukocyte-reduced suspended red blood cells(L-SRBC) and MAP resuspended leukocyte-reduced washed red blood cells(MAP-WRBC). On the 2nd, 7th, 14th, 21st, 28th and 35th day of blood sampling, sterility test, pH value, CD47 protein average fluorescence intensity and PS protein positive rate were detected.Results The mean fluorescence intensity of CD47 was significantly decreased on days 2, 7, 14 and 21 in MAP-WRBC compared with L-SRBC on days 35(457.35±81.67, 421.93±54.01, 304.03±26.08, 288.79±107.07 vs 209.81±26.17, P < 0.05). The positive rate of PS protein was significantly decreased on the 2nd, 7th, 14th, 21st and 35th days in MAP-WRBC compared with L-SRBC on the 35th(0.08 ±0.13, 0.07±0.13, 0.03±0.02, 4.14±1.53, 1.34±0.56 vs 2.29±0.31, P < 0.05). The pH value of MAP-WRBC was significantly lower than that of L-SRBC(day 2: 6.82±0.04 vs 6.9±0.09, day 7: 6.64±0.04 vs 6.72±0.07, day 14: 6.57±0.03 vs 6.66±0.08, day 21: 6.49±0.02 vs 6.58±0.04, day 35: 6.45±0.02 vs 6.52±0.03, P < 0.05; day 28: 6.46±0.02 vs 6.55±0.03, P < 0.01).Conclusion Multiple addition of low pH solution can aggravate the damage of erythrocyte membrane, decrease the expression of CD47 protein, and increase PS protein ectropion of MAP-WRBC. The damage of erythrocyte membrane at 21 days was lower than that at the end of the preservation period of L-SRBC. It is suggested that 21 days should be the preservation period of MAP-WRBC. -
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表 1 2组CD47平均荧光强度比较
X±S 组别 第2天 第7天 第14天 第21天 第28天 第35天 去白细胞洗涤红细胞 457.35±81.67 421.93±54.01 304.03±26.08 288.79±107.07 235.46±45.17 220.95±31.40 去白细胞悬浮红细胞 209.81±26.17 P < 0.01 < 0.01 < 0.01 < 0.05 > 0.05 > 0.05 
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表 2 2组AV标记阳性率比较
X±S 组别 第2天 第7天 第14天 第21天 第28天 第35天 去白细胞洗涤红细胞 0.08±0.13 0.07±0.13 0.03±0.02 1.34±0.52 2.19±0.56 4.14±1.53 去白细胞悬浮红细胞 2.29±0.31 P < 0.01 < 0.01 < 0.01 < 0.05 > 0.05 < 0.01
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表 3 2组pH值比较
X±S 组别 第2天 第7天 第14天 第21天 第28天 第35天 去白细胞洗涤红细胞 6.82±0.04 6.64±0.04 6.57±0.03 6.49±0.02 6.46±0.02 6.45±0.02 去白细胞悬浮红细胞 6.90±0.09 6.72±0.07 6.66±0.08 6.58±0.04 6.55±0.03 6.52±0.03 P < 0.05 < 0.05 < 0.05 < 0.05 < 0.01 < 0.05
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[1] 郑井滨, 张丽, 姚美辰, 等. 添加液去白洗涤红细胞在保存中的质量观察[J]. 临床输血与检验, 2020, 22(2): 122-126. https://www.cnki.com.cn/Article/CJFDTOTAL-LSXY202002003.htm
[2] 强书毓, 董晏榕, 张玉龙, 等. 储存红细胞体内外质量指标相关性分析研究[J]. 临床输血与检验, 2022, 24(2): 182-188. https://www.cnki.com.cn/Article/CJFDTOTAL-LSXY202202009.htm
[3] 胡连训. 高海拔地区洗涤红细胞经MAP液悬浮后的质量观察[J]. 青海医药杂志, 2016, 46(6): 57-58. https://www.cnki.com.cn/Article/CJFDTOTAL-QHYZ201606026.htm
[4] 段志倩, 石洁, 郑建勇, 等. MAP保存洗涤红细胞质量评价[J]. 临床血液学杂志, 2016, 29(2): 144-145. https://www.cnki.com.cn/Article/CJFDTOTAL-LCXZ201602020.htm
[5] 何亚琴, 吴敏, 马天虹, 等. 红细胞保存液悬浮洗涤红细胞的动态质量观察[J]. 临床血液学杂志, 2015, 28(10): 841-843. https://www.cnki.com.cn/Article/CJFDTOTAL-LCXZ201510005.htm
[6] 王伟, 严京梅, 王淑君, 等. 红细胞代谢产物及其表面分子对巨噬细胞免疫功能影响的研究进展[J]. 中国输血杂志, 2019, 32(10): 1076-1079. https://www.cnki.com.cn/Article/CJFDTOTAL-BLOO201910035.htm
[7] Klei TR, Meinderts SM, van den Berg TK, et al. From the Cradle to the Grave: The Role of Macrophages in Erythropoiesis and Erythrophagocytosis[J]. Front Immunol, 2017, 8: 73. http://www.onacademic.com/detail/journal_1000040529000810_df4b.html
[8] Arias CF, Arias CF. How do red blood cells know when to die?[J]. R Soc Open Sci, 2017, 4(4): 160850. http://rsos.royalsocietypublishing.org/content/4/4/160850.reviewer-comments.pdf
[9] 彭明喜, 戴梦玲, 许德义, 等. 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[J]. 临床血液学杂志, 2019, 32(8): 571-574. https://www.cnki.com.cn/Article/CJFDTOTAL-LCXZ201908001.htm
[10] Barkur S, Lukose J, Chidangil S. Probing Nanoparticle-Cell Interaction Using Micro-Raman Spectroscopy: Silver and Gold Nanoparticle-Induced Stress Effects on Optically Trapped Live Red Blood Cells[J]. ACS Omega, 2020, 5(3): 1439-1447. http://pubmed.ncbi.nlm.nih.gov/32010816/
[11] Bouchard BA, Orfeo T, Keith HN, et al. Microparticles formed during storage of red blood cell units support thrombin generation[J]. J Trauma Acute Care Surg, 2018, 84(4): 598-605. http://www.onacademic.com/detail/journal_1000040145224010_4316.html
[12] Chang AL, Kim Y, Seitz AP, Schuster RM, Pritts TA. pH modulation ameliorates the red blood cell storage lesion in a murine model of transfusion[J]. J Surg Res, 2017, 212: 54-59. http://www.onacademic.com/detail/journal_1000039788678710_dc2d.html
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