【作者】 李丹；

【导师】 肖子辉；

【作者基本信息】 中南大学 ， 病理学与病理生理学， 2014， 硕士

【摘要】 目的：脓毒症常常伴随不同程度的凝血功能异常,血小板在炎症和凝血反应过程中均发挥着重要作用。以往研究显示,在大鼠内毒素血症模型中,氯吡格雷可以改善肝肺组织的病理改变,减轻促炎介质IL-6和TNF-α及肝肺组织中HMGB1的水平。本研究以SD大鼠作为研究对象,经盲肠结扎穿刺(CLP)复制脓毒症模型,探讨血小板拮抗剂氯吡格雷对脓毒症大鼠生存率、血小板-白细胞相互作用和白细胞组织因子表达及内皮细胞激活的影响。方法：将雄性SD大鼠随机分为四组：1.假手术组；2.假手术+氯吡格雷组；3.CLP组；4.CLP+氯吡格雷组。观察术后7天内各组大鼠的生存率；并采用流式细胞术检测各组大鼠血小板和白细胞激活,血小板-白细胞相互作用及白细胞表面组织因子的表达；采用Elisa试剂盒分别检测大鼠ICAM-1, E-Selectin, PSGL-1的血浆水平。结果：(1)生存率：CLP组大鼠生存率与假手术组/假手术+氯吡格雷组相比显著下降(p<0.05)；而CLP+氯吡格雷组大鼠生存率与CLP组相比显著升高(p<0.05)。(2)流式细胞术检测：术后24小时,与假手术组/假手术+氯吡格雷组相比,CLP组大鼠血小板、白细胞激活,血小板-白细胞相互作用及白细胞组织因子表达明显增高(p<0.05)；而与CLP组相比,CLP+氯吡格雷组大鼠血小板和白细胞激活,血小板-白细胞相互作用及白细胞组织因子表达显著降低(p<0.05)。(3)Elisa检测：术后12小时,CLP组大鼠血浆中ICAM-1, E-Selectin的水平与假手术组/假手术+氯吡格雷组相比显著升高(p＜0.05); CLP+氯吡格雷组大鼠血浆中ICAM-1, E-Selectin的水平均显著低于CLP组(p<0.05)；但各组大鼠血浆PSGL-1水平没有明显差别(p>0.05)。术后24小时,CLP组大鼠血浆中ICAM-1、PSGL-1的水平与假手术组／假手术+氯吡格雷组相比显著升高(p＜0.05); CLP+氯吡格雷组大鼠血浆中ICAM-1、PSGL-1水平显著低于CLP组(p<0.05),而各组大鼠血浆E-Selectin水平没有明显差别(p>0.05)。结论：血小板拮抗剂氯吡格雷显著抑制脓毒症所致的血小板、白细胞和内皮细胞的激活及血小板-白细胞粘附和白细胞组织因子表达的上调,并提高脓毒症大鼠的生存率。更多还原

【Abstract】 Objective:Sepsis is often associated with coagulation abnormalities. Platelets play an important role both in inflammatory and coagulation processes. Previous studies showed that in a rat model of endotoxemia, clopidogrel could attenuate the histological changes of liver and lung tissue, and reduce the production of cytokines (IL-6and TNF-a) and the expression of the high mobility group box1(HMGB1) in liver and lung tissues. In this study, SD rats were recruited to produce an animal model of sepsis and farther explore the impact of clopidogrel on the activation of platelets, leukocytes and endothelial cells, the interaction of platelets with leukocytes, the expression of tissue factor on leukocytes and survival percentage of rats during the sepsis.Method:A rat model of sepsis was established by cecal ligation and puncture (CLP). SD rats were randomly divided into4groups:(1) sham-operated group (Sham);(2)sham-operated group with clopidogrel sulfate treatment (Sham+CS);(3) CLP group (CLP);(4) CLP group with clopidogrel sulfate treatment (CLP+CS). The survival percentages were observed in each group within7days after operation. The flow cytometric analyses were employed to detect the activation of platelets and leukocytes, platelet-leukocyte coaggregates and tissue factor (TF) expression on leukocytes, and Elisa kits to detect the plasm content of ICAM-1, E-Selectin and PSGL-1.Results:(1) Compared with the Sham group, the mortality of the CLP group was markedly increased (p<0.05), and CS significantly reduced the mortality (p<0.05).(2) At24h after operation, compared with the Sham group, the expression of CD61on platelets and Mac-1on leukocytes, the percentage of platelet-leukocyte coaggregates and expression of tissue factor on leukocytes in CLP group were significantly enhanced; and all these values were significantly reduced in CLP+CS group (all p＜0.05).(3) At12h after operation, compared with Sham group, the plasma content of ICAM-1, E-Selectin of CLP group were significantly increased (p<0.05), and CS markedly decreased the upregulation of ICAM-1and E-Selectin (p<0.05). However, there was no significant effect of CS on the PSGL-1(p>0.05).At24h after operation, CS markedly decreased the upregulation of ICAM-1and PSGL-1(p<0.05). However, no significant effect of CS was found in the E-Selectin (p>0.05).Conclusion:1.During sepsis, CS could inhibit the activation of platelets, leukocytes and endothelial cells, the interaction of platelets with leukocytes and the expression of tissue factor on leukocytes;2.During sepsis, CS could improve the mortality of rats.更多还原