Xie Rufeng, Yang Yiming, Jiang Xueyu, Gao Li, Sun Juan, Yang Jie
Blood Engineering Laboratory, Shanghai Blood Center, Shanghai, China.
Blood Engineering Laboratory, Shanghai Blood Center, Shanghai, China.
Hematol Transfus Cell Ther. 2024 Dec;46 Suppl 6(Suppl 6):S272-S283. doi: 10.1016/j.htct.2024.09.2479. Epub 2024 Oct 18.
It is known that the rapid clearance of cold-stored platelets is attributed to various storage lesions, including an abnormal increase in reactive oxygen species when platelets are exposed to cold temperatures. As an antioxidant, N-acetylcysteine exhibits some significant effects on scavenging various reactive oxygen species and inhibiting cell damage and apoptosis.
This study aimed to investigate the effects of N-acetylcysteine on reducing reactive oxygen species production and protecting cold-stored platelets from phagocytosis and clearance, and to determine the optimal concentration of N-acetylcysteine.
Platelet concentrates were divided into three groups: room-temperature-stored platelets, cold-stored platelets, and cold-stored platelets with the addition of different concentrations of N-acetylcysteine. After five days of storage, reactive oxygen species production, lipid peroxidation levels, activation marker expressions, GPIb/ɑ desialylation with exposure of glycan residues and other quality parameters of platelets were measured and compared between the groups. Phagocytosis of platelets was detected by phorbol 12-myristate 13-acetate-activated THP-1 or Hep G2 cells. Moreover, the recovery of infused platelets was measured in severe combined immunodeficient mice at different timepoints.
After 5 days of storage, cytoplasmic reactive oxygen species significantly increased in chilled compared to non-chilled platelets; they were notably reduced with the addition of N-acetylcysteine, particularly at a concentration of 5 mM. Compared with chilled platelets, the P-selectin and phosphatidylserine expressions, as well as exposure of GPIb/ɑ glycan residues, were significantly reduced with 5 mM of N-acetylcysteine. Phagocytosis of platelets by THP-1 or Hep G2 cells was significantly lower in 5 mM of N-acetylcysteine compared to cold-stored platelets without N-acetylcysteine.
This study demonstrated correlations between reactive oxygen species production and their pro-oxidant effects on platelet clearance after cold storage. The addition of N-acetylcysteine at an appropriate concentration do not only protects chilled platelets from storage lesions caused by reactive oxygen species overproduction but also prevents platelet phagocytosis in vitro and clearance in vivo, thereby extending circulating time.
已知冷藏血小板的快速清除归因于各种储存损伤，包括血小板暴露于低温时活性氧的异常增加。作为一种抗氧化剂，N-乙酰半胱氨酸在清除各种活性氧以及抑制细胞损伤和凋亡方面表现出一些显著作用。
本研究旨在探讨N-乙酰半胱氨酸对减少活性氧产生以及保护冷藏血小板免受吞噬和清除的作用，并确定N-乙酰半胱氨酸的最佳浓度。
将血小板浓缩物分为三组：室温储存的血小板、冷藏血小板以及添加不同浓度N-乙酰半胱氨酸的冷藏血小板。储存五天后，测量并比较各组血小板的活性氧产生、脂质过氧化水平、活化标志物表达、糖蛋白Ib/α去唾液酸化及聚糖残基暴露情况等质量参数。通过佛波醇1-肉豆蔻酸酯13-乙酸酯激活的THP-1或Hep G2细胞检测血小板的吞噬作用。此外，在不同时间点测量重度联合免疫缺陷小鼠体内输注血小板的回收率。
储存5天后，与未冷藏的血小板相比，冷藏血小板的细胞质活性氧显著增加；添加N-乙酰半胱氨酸后活性氧显著减少，尤其是在浓度为5 mM时。与冷藏血小板相比，5 mM的N-乙酰半胱氨酸可显著降低P-选择素和磷脂酰丝氨酸的表达以及糖蛋白Ib/α聚糖残基的暴露。与未添加N-乙酰半胱氨酸的冷藏血小板相比，5 mM的N-乙酰半胱氨酸组中THP-1或Hep G2细胞对血小板的吞噬作用显著降低。
本研究证明了活性氧产生与其对冷藏后血小板清除的促氧化作用之间的相关性。添加适当浓度的N-乙酰半胱氨酸不仅可保护冷藏血小板免受活性氧过度产生所致的储存损伤，还可在体外防止血小板吞噬并在体内防止血小板清除，从而延长循环时间。
