微粒和血细胞在脓毒症高凝状态中的作用

Role of the Microparticle(MP) and PS Exposure of Erythrocytes and Platelets in the Procoagulant State of Sepsis

目的:探讨血小板和红细胞磷脂酰丝氨酸(PS)暴露和循环微粒(MPs)在脓毒症高凝状态中的作用。方法:集10例诊断为脓毒症的患者及10例健康体检者的新鲜血;离心分离出红细胞、富血小板血浆及MPs;应用Alexa-Fluro 488-乳粘素/CD 41a/CD14、Alexa-Fluro 647-CD 235a/CD 31/CD 45标定红细胞、血小板、内皮细胞、白细胞及单核细胞来源的MPs;流式细胞仪测定红细胞和血小板的PS外翻、MPs的数量;血凝仪测定相应凝血时间。结果:相对于健康人,脓毒症患者PS外翻红细胞(3.53%vs0.53%,P<0.05)和血小板(3.00%vs 0.45%,P<0.05)、MPs总数(4830/μL vs 2861/μL,P<0.05)及各亚型MPs均明显升高;红细胞(238.4s vs 346.8s,P<0.05)、血小板(209.1s vs 335.9s,P<0.05)及MPs(207.1s vs 318.2s,P<0.05)的凝血时间缩短。MP数、PS+红细胞/血小板比例分别与各自凝血时间成负相关(MP:r=-0.592,P=0.006;红细胞:r=-0.565,P=0.009;血小板:r=-0.641,P=0.002)。结论:红细胞、血小板的PS暴露及循环MPs和脓毒症高凝状态的发生相关,可能对患者凝血功能检测和抗凝治疗有一定的意义。

Objective： To investigate the role of microparticle （MP） and phosphatidylserine （PS） exposure of erythrocytes and platelets in the procoagulant state in sepsis. Methods： Peripheral blood samples were collected from 10 patients with newly diagnosed sepsis and 10 matched healthy subjects. Erythrocytes, platelets, and MPs were obtained by centrifugation. Alexa-Fluro 488-1actadherin/CD 41a/CD 14 and Alexa-Fluro 647-CD 235a/CD 31/CD 45 were used to label MPs derived from platelets, monocytes, erythrocytes, endothelial cells, and leukocytes. PS exposure of platelets and erythrocytes was detected with Alexa-Fluro 488-1actadherin. Analysis of MP release and PS exposure on erythrocytes and platelets were performed with a flow cytometer. Procoagulant activity （PCA） was determined by clotting time. Results： Compared with healthy subjects, patients with sepsis had markedly increased MP release （4830/μLvs 2861/μL, P〈0.05） and PS exposure on erythrocytes （3.53% vs 0.53%, P〈0.05） and platelets （3.00% vs 0.45%, P〈0.05）. Relevant PCA was significantly increased as determined by shortened clotting time （erythrocytes： 238.4s vs 346.8s, P〈0.05; platelets： 209. l s vs 335.9s, P〈0.05; MPs： 207.1s vs 318.2s, P〈0.05）. The level of circulating MPs, PS＋ erythrocytes %, and PS＋ platelets% was negatively correlated with its own relevant clotting time （MP： r= -0.592, P=0.006; erythrocytes： r= -0.565, P=0.009; platelets： r= -0.641, P=0.002）. Conclusions： Increased MPs and PS＋ erythrocytes/platelets are associated with the procoagulant state in patients with sepsis, and may have clinical significance in evaluating and predicting the effect of anticoagnlant therapy.