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海藻糖:一种用于浓缩血小板的添加剂溶液,可在4℃储存期间保护血小板免于凋亡和清除。

Trehalose An Additive Solution for Platelet Concentrate to Protect Platelets from Apoptosis and Clearance during Their Storage at 4°C.

作者信息

Baghdadi Vahid, Ranjbaran Reza, Yari Fatemeh, Rafiee Mohammad Hessam

机构信息

Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.

Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.

出版信息

Cell J. 2022 Feb;24(2):69-75. doi: 10.22074/cellj.2022.7886.

Abstract

OBJECTIVE

Although cold storage of platelets (PLTs) could decrease the risk of bacterial growth, it could affect on the PLTs viability and hemostatic function. At cold temperatures, trehalose can be used to substitute water, inhibit the solid-liquid transition phase of the PLT membrane, and stop Glycoprotein Ibα (GPIbα) polymerization. In this study, we evaluated the potential of trehalose for reducing the negative effects of cold storage on the apoptosis and the clearance rates of PLTs after long-term storage at cold.

MATERIALS AND METHODS

In this experimental study, PLT concentrates (PCs) were maintained for five days in the different circumstances. PLTs were subsequently counted by using an automated hematology analyzer. Also water-soluble tetrazolium salt (WST-1) assay was performed to estimate the viability of PLTs. The activity of lactate dehydrogenase enzyme (LDH) was determined by a biochemical analyzer. And human active caspase-3 levels were measured by using enzyme-linked immunosorbent assay (ELISA) method. Also, we applied flow cytometry technique.

RESULTS

PLTs count and viability were higher, while LDH amount was lower in trehalose-treated PLTs when compared with two other groups (P=0.03). The highest increase in the amount of caspase-3 levels in the PLTs was observed at 4°C. However, trehalose-treated and 4°C PLTs had a lower amount of active caspase-3 in comparison with 4°C PLTs. The level of PS expression on PLTs was lower in the trehalose-treated PLTs in compared with the two other groups (P=0.03). PLTs ingestion by HepG2 cells was enhanced in the 4°C-stored PLTs. However, the ingestion rate was significantly reduced in the trehalose-treated PLTs on day 5 of storage (P=0.03).

CONCLUSION

Trehalose can moderate the effects of cold temperature on the apoptosis, viability, and the survival rate of PLTs. It also decreases the ingestion rate of refrigerated PLTs .

摘要

目的

尽管血小板(PLT)冷藏可降低细菌生长风险,但会影响PLT的活力和止血功能。在低温下,海藻糖可用于替代水分,抑制PLT膜的固液转变阶段,并阻止糖蛋白Ibα(GPIbα)聚合。在本研究中,我们评估了海藻糖在减轻冷藏对PLT长期冷藏后凋亡及清除率负面影响方面的潜力。

材料与方法

在本实验研究中,PLT浓缩物(PC)在不同条件下保存5天。随后使用自动血液分析仪对PLT进行计数。还进行了水溶性四氮唑盐(WST-1)检测以评估PLT的活力。通过生化分析仪测定乳酸脱氢酶(LDH)的活性。并使用酶联免疫吸附测定(ELISA)法测量人活性半胱天冬酶-3水平。此外,我们应用了流式细胞术技术。

结果

与其他两组相比,海藻糖处理的PLT中PLT计数和活力更高,而LDH含量更低(P = 0.03)。在4°C时观察到PLT中半胱天冬酶-3水平的升高幅度最大。然而,与4°C保存的PLT相比,海藻糖处理的4°C PLT中活性半胱天冬酶-3的含量更低。与其他两组相比,海藻糖处理的PLT中PLT上磷脂酰丝氨酸(PS)的表达水平更低(P = 0.03)。4°C保存的PLT中HepG2细胞对PLT的摄取增强。然而,在保存第5天时,海藻糖处理的PLT的摄取率显著降低(P = 0.03)。

结论

海藻糖可减轻低温对PLT凋亡、活力和存活率的影响。它还降低了冷藏PLT的摄取率。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/85f0/8918273/e1cdf3f0cabe/Cell-J-24-69-g01.jpg

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