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不同方法调整血小板数量对血小板聚集功能检测的影响

Adjusting Platelet Counts for Platelet Aggregation Tests

  • 摘要: 目的寻求更好的方法调整富血小板血浆(PRP)中的血小板数量,以满足光透射血小板聚集检测(LTA)对样本的质量要求。方法收集18~50岁健康人血样36例。分别以乏血小板血浆(PPP)和生理盐水(PS)调整PRP中血小板数量,调整(稀释)倍数为1.5倍、2倍、2.5倍和3倍,以终浓度5 μmol/L二磷酸腺苷(ADP)、0.5 mmol/L花生四烯酸(ARA)、2 μg/mL胶原(COL)、5 μmol/L肾上腺素(EPI)以及1.2 mg/mL瑞斯托霉素(RIS)为诱导剂,测定调整前后PRP的血小板最大聚集率(MA)的差异。用PRP离心制备的PPP(离心PRP-获取-PPP),比较其与传统方法(离心全血-获取-PPP)对RIS诱导血小板聚集的影响。结果当ADP、ARA或EPI为激活剂时,PS各调整倍数的PRP与原PRP相比,其MA值的差异无统计学意义(P>0.05);而PPP各调整倍数的PRP与原PRP相比,其MA值均下降,在调整倍数为2~3倍时,差异有统计学意义(P<0.05)。当激活剂为RIS时,PS各调整倍数的PRP 与原PRP相比,其MA值均下降,且差异有统计学意义(P<0.05);而PPP各调整倍数的PRP与原PRP相比,其MA值的差异无统计学意义(P>0.05)。当激活剂为COL时,PS和PPP各调整倍数的PRP与原PRP相比,其MA值的差异均无统计学意义(P>0.05)。全血或PRP离心制备的PPP,均不影响RIS诱导血小板聚集的MA值(P>0.05)。结论以ADP、ARA、COL或EPI为诱导剂检测MA时,推荐使用PS调整血小板数量;而以RIS为诱导剂时,应使用自身PPP调整血小板数量;2 100×g,5 min条件下用传统离心全血制备的PPP方便可行。

     

    Abstract: ObjectiveTo explore a better method to adjust platelet counts for light transmission aggregometry (LTA). MethodsBlood samples from 36 healthy participants aged from 18 to 50 yr. were collected.Platelet-rich plasma (PRP) was diluted using platelet-poor plasma (PPP) and physiological saline (PS), respectively, in a ratio of 1.5, 2, 2.5 and 3 times. Platelet aggregation was induced by adenosine diphosphate (ADP), arachidonic acid (ARA), collagen (COL), epinephrine (EPI), or ristocetin (RIS). The maximal aggregation rates (MAs) of different approaches were compared. We also compared the MAs induced by RIS between PRP-obtained-PPP and whole blood-obtained-PPP (2 100×g, 5 min). ResultsCompared with the original PRP, the MAs induced by ADP, ARA, and EPI decreased in PPP-adjusted PRP (significant at 2-3 times dilution ratio, P<0.05), but not in PS-adjusted PRP (P>0.05). The MA induced by RIS decreased in PS-adjusted PRP (significant at all dilution ratios, P<0.05), but not in PPP-adjusted PRP (P>0.05). No changes in the MA induced by COL were found in PS-adjusted PRP and PPP-adjusted PRP (P>0.05). Whole blood-obtained-PPP (2 100×g, 5 min) had the same MA induced by ristocetin compared with PRP-obtained-PPP (P>0.05). ConclusionPS is recommended for adjusting platelets counts for platelet aggregation induced by ADP, ARA, COL and EPI. Whole blood-obtained-PPP (2 100 ×g, 5 min) is recommended for RIS-induced aggregation as a matter of convenience.

     

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