活化血小板对内皮细胞白细胞介素IL-1β和IL-6表达的影响

发布时间：2018-05-01 01:39

  本文选题：血小板活化 + 炎症反应　； 参考：《天津医科大学》2007年硕士论文

【摘要】： 目的 通过分析血小板受ADP诱导剂活化后对体外培养的人脐静脉内皮细胞株表达炎症细胞因子白介素IL-1β和IL-6的影响，探讨活化血小板在炎症反应性疾病中的作用。 方法 1取30例健康人外周血，根据Ahnadi CE等的方法制备血小板悬液，用最终浓度为0.8μmol／l的ADP与血小板作用20min，诱导血小板活化。用Advia120血细胞分析仪检测各样本ADP处理前后血小板平均内含物浓度(MPC)的变化，评价血小板活化水平。 2复苏冰冻的ECV304人脐静脉内皮细胞(HUVECs)株，传代培养。当细胞生长良好时，吸去培养液进行血小板刺激实验。采集健康人外周血制备成混合血小板悬液，分以下四个实验组：(1)活化血小板组：终浓度为0.8μmol／l的ADP与血小板室温下作用20 min后再与HUVECs共同孵育。(2)静息血小板组：血小板悬液室温下静置20 min后与HUVECs共同孵育。(3)ADP对照组(4)空白细胞对照组。各组细胞在5％CO_2 37℃条件下孵育12小时，，用TRNzol提取总RNA，再以RT-PCR技术检测内皮细胞白介素IL-1β和IL-6mRNA的表达。 结果 1最终浓度为0.8μmol／l的ADP刺激血小板20min前后MPC水平经配对t检验，t=32.24，P＜0.01，差别有显著性，说明该实验条件可在体外刺激血小板活化。 2活化的血小板与HUVECs共同培养后，可刺激内皮细胞表达炎症细胞因子IL-1β和IL-6mRNA，而静息的血小板以及相同含量的ADP均不能刺激内皮细胞IL-1β和IL-6mRNA的表达。 结论 活化的血小板可通过某种途径诱导血管内皮细胞表达炎性细胞因子IL-1β和IL-6，进而参与炎症反应。这种血小板-内皮反应有可能是某些炎症反应性疾病的一条重要病理机制。
[Abstract]:Purpose By analyzing the effect of platelet activation by ADP inducer on the expression of inflammatory cytokines interleukin IL-1 尾 and IL-6 in human umbilical vein endothelial cell lines in vitro, the role of activated platelets in inflammatory response disease was investigated. Method 1Peripheral blood samples were collected from 30 healthy people. Platelet suspension was prepared by Ahnadi CE et al. Platelet activation was induced by the action of 0.8 渭 mol/l ADP with platelet for 20 min. The changes of mean platelet concentration before and after ADP treatment were detected by Advia120 hematology analyzer to evaluate the platelet activation level. 2 ECV304 human umbilical vein endothelial cells (HUVECs) were subcultured. When the cells grew well, the platelets were stimulated by sucking the culture medium. A mixture of platelet suspensions was prepared from the peripheral blood of healthy people. Four experimental groups: activated platelets group: ADP with final concentration of 0.8 渭 mol/l was incubated with HUVECs for 20 min at room temperature and then incubated with HUVECs at room temperature. The rest platelet group: platelet suspension was incubated with HUVECs at room temperature for 20 min and then incubated with HUVECs. Control group 4) blank cell control group. The cells in each group were incubated at 5%CO_2 37 鈩

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