查看更多>>摘要：目的 评价肝组织ERBB2相互作用蛋白(Erbin)在脓毒症小鼠凝血功能中的作用。 方法 SPF级健康雄性C57BL/6小鼠和Erbin基因敲除小鼠各30只，8～10周龄，体质量20～30 g，采用随机数字表法分为野生型＋假手术组(WT＋Sham组)、野生型＋脓毒症组(WT＋SEP组)、Erbin基因敲除＋假手术组(EKO＋Sham组)和Erbin基因敲除＋脓毒症组(EKO＋SEP组)，每组15只。采用盲肠结扎穿孔法制备小鼠脓毒症模型，术后24 h时采集眼球血样，随后处死小鼠取肝组织，HE染色法观察肝组织病理学结果并进行评分，比色法测定血浆ALT和AST活性，Western blot法检测Erbin和组织因子(TF)表达，qPCR法检测组织型纤溶酶原激活物(t-PA)和Fib的mRNA表达，采用全自动凝血分析仪检测PT、APTT、凝血酶时间(TT)和Fib浓度，采用ELISA法测定血浆TF和IL-6浓度。 结果 与WT＋Sham组比较，WT＋SEP组小鼠肝组织病理学损伤评分升高，TF表达、t-PA和FGA的mRNA表达上调，PT、APTT和TT延长，血浆Fib浓度、ALT、AST活性、TF和IL-6浓度升高(P<0.05)；与WT＋SEP组比较，EKO＋SEP组小鼠肝组织病理学损伤评分升高，肝组织TF表达、t-PA和FGA的mRNA表达上调，Erbin表达下调，PT、APTT和TT延长，血浆Fib浓度、ALT、AST活性、TF和IL-6浓度升高(P<0.05)。 结论 肝组织Erbin通过抑制TF表达上调对脓毒症小鼠凝血功能发挥内源性保护作用。 Objective To evaluate the role of ERBB2 interacting protein (Erbin)in liver tissues in blood coagulation of septic mice. Methods Thirty SPF healthy male C57BL/6 mice and 30 Erbin knockout mice, aged 8-10 weeks, weighing 20-30 g, were divided into wild-type+ sham operation group (WT+ Sham group), wild-type+ sepsis group (WT+ SEP group), Erbin gene knockout+ sham operation group (EKO+ Sham group) and Erbin gene knockout+ sepsis group (EKO+ SEP group) by a random number table method, with 15 animals in each group. The mouse sepsis model was prepared by the cecal ligation and perforation method in anesthetized animals. Eye blood samples were collected at 24 h after surgery and liver tissues were obtained for microscopic examination of histopathological changes (by HE staining) which were scored and for determination of plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities (by colorimetry), expression of Erbin and tissue factor (TF) (by Western blot), expression of tissue plasminogen activator (t-PA) and fibrinogen (Fib)mRNA (by quantitative polymerase chain reaction), concentrations of PT, APTT, thrombin time (TT) and Fib (by automatic coagulation analyzer), and plasma TF and interleukin-6 (IL-6) concentrations (by enzyme-linked immunosorbent assay). Results Compared with WT+ Sham group, the lung injury score was significantly increased, the expression of TF, t-PA mRNA and FGA mRNA was up-regulated, PT, APTT and TT were prolonged, the plasma Fib concentration was increased, and the activities of ALT and AST and concentrations of TF and IL-6 in plasma were increased in WT+ SEP group (P<0.05). Compared with WT+ SEP group, the lung injury score was significantly increased, the expression of TF, t-PA mRNA and FGA mRNA was up-regulated, PT, APTT and TT were prolonged, the plasma Fib concentration was increased, and the activities of ALT and AST and concentrations of TF and IL-6 in plasma were increased in EKO+ SEP group (P<0.05). Conclusions Erbin in liver tissues exerts an endogenous protective effect on blood coagulation by inhibiting the up-regulation of TF expression in septic mice.
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