本文選題：血小板活化 + 炎癥反應��； 參考：《天津醫(yī)科大學》2007年碩士論文

【摘要】： 目的 通過分析血小板受ADP誘導劑活化后對體外培養(yǎng)的人臍靜脈內(nèi)皮細胞株表達炎癥細胞因子白介素IL-1β和IL-6的影響，探討活化血小板在炎癥反應性疾病中的作用。 方法 1取30例健康人外周血，根據(jù)Ahnadi CE等的方法制備血小板懸液，用最終濃度為0.8μmol／l的ADP與血小板作用20min，誘導血小板活化。用Advia120血細胞分析儀檢測各樣本ADP處理前后血小板平均內(nèi)含物濃度(MPC)的變化，評價血小板活化水平。 2復蘇冰凍的ECV304人臍靜脈內(nèi)皮細胞(HUVECs)株，傳代培養(yǎng)。當細胞生長良好時，吸去培養(yǎng)液進行血小板刺激實驗。采集健康人外周血制備成混合血小板懸液，分以下四個實驗組：(1)活化血小板組：終濃度為0.8μmol／l的ADP與血小板室溫下作用20 min后再與HUVECs共同孵育。(2)靜息血小板組：血小板懸液室溫下靜置20 min后與HUVECs共同孵育。(3)ADP對照組(4)空白細胞對照組。各組細胞在5％CO_2 37℃條件下孵育12小時，，用TRNzol提取總RNA，再以RT-PCR技術(shù)檢測內(nèi)皮細胞白介素IL-1β和IL-6mRNA的表達。 結(jié)果 1最終濃度為0.8μmol／l的ADP刺激血小板20min前后MPC水平經(jīng)配對t檢驗，t=32.24，P＜0.01，差別有顯著性，說明該實驗條件可在體外刺激血小板活化。 2活化的血小板與HUVECs共同培養(yǎng)后，可刺激內(nèi)皮細胞表達炎癥細胞因子IL-1β和IL-6mRNA，而靜息的血小板以及相同含量的ADP均不能刺激內(nèi)皮細胞IL-1β和IL-6mRNA的表達。 結(jié)論 活化的血小板可通過某種途徑誘導血管內(nèi)皮細胞表達炎性細胞因子IL-1β和IL-6，進而參與炎癥反應。這種血小板-內(nèi)皮反應有可能是某些炎癥反應性疾病的一條重要病理機制。
[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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