Institute for Clinical Chemistry and Laboratory Medicine, Regensburg, Germany.
Transfusion. 2014 Sep;54(9):2301-14. doi: 10.1111/trf.12640. Epub 2014 Jun 10.
Stored platelet (PLT) concentrates (PLCs) for transfusion develop a PLT storage lesion (PSL), decreasing PLT viability and function with profound lipidomic changes and PLT extracellular vesicle (PL-EV) release. High-density lipoprotein 3 (HDL3 ) improves PLT homeostasis through silencing effects on PLT activation in vivo. This prompted us to investigate HDL3 and apolipoprotein A-I (apoA-I) as PSL-antagonizing agents.
Healthy donor PLCs were split into low-volume standard PLC storage bags and incubated with native (n)HDL3 or apoA-I from plasma ethanol fractionation (precipitate IV) for 5 days under standard blood banking conditions. Flow cytometry, Born aggregometry, and lipid mass spectrometry were carried out to analyze PL-EV release, PLT aggregation, agonist-induced PLT surface marker expression, and PLT and plasma lipid compositions.
Compared to control, added nHDL3 and apoA-I significantly reduced PL-EV release by up to -62% during 5 days, correlating with the added apoA-I concentration. At the lipid level, nHDL3 and apoA-I antagonized PLT lipid loss (+12%) and decreased cholesteryl ester (CE)/free cholesterol (FC) ratios (-69%), whereas in plasma polyunsaturated/saturated CE ratios increased (+3%) and CE 16:0/20:4 ratios decreased (-5%). Administration of nHDL3 increased PLT bis(monoacylglycero)phosphate/phosphatidylglycerol (+102%) and phosphatidic acid/lysophosphatidic acid (+255%) ratios and improved thrombin receptor-activating peptide 6-induced PLT aggregation (+5%).
nHDL3 and apoA-I improve PLT membrane homeostasis and intracellular lipid processing and increase CE efflux, antagonizing PSL-related reduction in PLT viability and function and PL-EV release. We suggest uptake and catabolism of nHDL3 into the PLT open canalicular system. As supplement in PLCs, nHDL3 or apoA-I from Fraction IV of plasma ethanol fractionation have the potential to improve PLC quality to prolong storage.
储存的血小板（PLT）浓缩物（PLCs）在输血过程中会发生 PLT 储存损伤（PSL），导致 PLT 活力和功能下降，并伴有深刻的脂质组学变化和 PLT 细胞外囊泡（PL-EV）释放。高密度脂蛋白 3（HDL3）通过在体内对 PLT 激活产生沉默作用来改善 PLT 动态平衡。这促使我们研究 HDL3 和载脂蛋白 A-I（apoA-I）作为 PSL 拮抗剂。
将健康供体的 PLC 分成小体积标准 PLC 储存袋，并在标准血库条件下用来自血浆乙醇分级沉淀（沉淀物 IV）的天然（n）HDL3 或 apoA-I 孵育 5 天。进行流式细胞术、Born 聚集测定和脂质质谱分析，以分析 PL-EV 释放、PLT 聚集、激动剂诱导的 PLT 表面标志物表达以及 PLT 和血浆脂质组成。
与对照组相比，添加的 nHDL3 和 apoA-I 在 5 天内显著减少了高达 62%的 PL-EV 释放，与添加的 apoA-I 浓度相关。在脂质水平上，nHDL3 和 apoA-I 拮抗 PLT 脂质损失（+12%）并降低胆固醇酯（CE）/游离胆固醇（FC）比值（-69%），而在血浆中，多不饱和/饱和 CE 比值增加（+3%），CE 16:0/20:4 比值降低（-5%）。nHDL3 的给药增加了 PLT 双（单酰基甘油）磷酸/磷脂酰甘油（+102%）和磷脂酸/溶血磷脂酸（+255%）比值，并改善了凝血酶受体激活肽 6 诱导的 PLT 聚集（+5%）。
nHDL3 和 apoA-I 改善了 PLT 膜动态平衡和细胞内脂质处理，并增加了 CE 外排，拮抗了与 PSL 相关的 PLT 活力和功能以及 PL-EV 释放的降低。我们建议 nHDL3 被摄取并代谢为 PLT 开放小管系统。作为 PLC 的补充，来自血浆乙醇分级沉淀的 Fraction IV 的 nHDL3 或 apoA-I 有可能改善 PLC 的质量，延长储存时间。
