枯否细胞通过组织因子对大鼠急性胰腺炎凝血系统变化的影响  

Effects of Kupffer cells on coagulation function of rats with acute pancreatitis through tissue factor

作　　者：幸定培[1] 龚建平[1] 冯虎翼[2] 吴念[2] 吴伟[1] 

机构地区：[1]重庆医科大学附属第二医院肝胆外科,重庆400010 [2]重庆市第五人民医院肝胆外科,重庆400062

出　　处：《重庆医科大学学报》2015年第3期329-334,共6页Journal of Chongqing Medical University

基　　金：国家自然科学基金青年基金资助项目(编号:81301618)

摘　　要：目的:观察大鼠急性胰腺炎(acute pancreatitis,AP)组织因子(tissue factor,TF)及凝血系统的变化,探讨枯否细胞(kupffer cells,KCs)通过组织因子对大鼠AP凝血系统的作用。方法:108只SD大鼠完全随机分为假手术(sham operation,SO)组、轻症胰腺炎(mild acute pancreatitis,MAP)组、重症胰腺炎(severe acute pancreatitis,SAP)组、SO+氯化钆(Gd Cl3)组、MAP+Gd Cl3组、SAP+Gd Cl3组。Gd Cl3预处理组在诱发AP模型前24 h予尾静脉注射Gd Cl3(10 mg/kg,0.1 ml),分别用1.5%和5%的牛黄胆酸钠胰胆管逆行注射诱导MAP和SAP模型。建模后或假手术组分别于术后6、12、24 h处死大鼠,检测其血清中淀粉酶及凝血项的变化;ELISA法检测血清中TF的含量;分离并培养KCs,采用Western blot法检测其TF的蛋白含量;免疫组化检测肝脏中TF蛋白的表达;HE染色检测胰腺组织病理损伤情况。结果:MAP和SAP组大鼠血清中淀粉酶含量明显高于SO组(P<0.01),与MAP+Gd Cl3组、SAP+Gd Cl3组各相应时间点比较无明显差异。同时间点MAP组和SAP组较SO组血清中TF含量明显升高,MAP+Gd Cl3组和SAP+Gd Cl3组较MAP组和SAP组明显降低(P<0.05)。MAP和SAP组大鼠肝脏中TF蛋白表达较SO组表达明显增加。MAP+Gd Cl3和SAP+Gd Cl3组KCs中TF蛋白含量较MAP组和SAP组明显降低(P<0.05)。SAP组较MAP和SO组大鼠血清凝血酶原时间(prothrombin time,PT)、活化部分凝血活酶时间(activated partial thromboplastin time,APTT)、纤维蛋白原浓度(fibrinogen,FIB)明显升高,SAP+Gd Cl3组较SAP明显降低(P<0.05)。大鼠胰腺病理切片显示,MAP组胰腺间质水肿,SAP组胰腺呈凝固性坏死,伴有大量炎症细胞浸润,MAP+Gd Cl3和SAP+Gd Cl3组分别较MAP和SAP组病理损伤明显减轻。结论:AP时KCs中TF表达明显增加,并能导致凝血象的改变,抑制KCs功能可有效降低AP时TF的表达,减少凝血象的改变,从一定程度上可降低AP的发生和发展。Objective：To observe the changes of tissue factor（TF）and coagulation function in rats with acute pancreatitis（AP）and to explore the role of Kupffer cells（KCs）in cruor function in AP rats via TF. Methods：Totally 108 SD rats are randomly divided into 6groups including sham operation（SO） group,mild acute pancreatitis（MAP） group,severe acute pancreatitis（SAP） group,SO ＋gadolinium chloride（Gdcl3）group,MAP＋Gdcl3group,SAP ＋Gdcl3group. Rats in Gdcl3 pretreatment group were injected with Gdcl3（10 mg/kg,0.1 ml）through the vena caudalis at 24 h before the establishment of AP model. Rats were retrograde infected with 1.5%or 5% of sodium taurocholate through bile duct to establish MAP model and SAP model. The rats were killed at 6,12,24 h after AP treatment or sham operation. Changes in serum amylase and coagulation makers were detected. The content of TF in serum was measured by ELISA. KCs was isolated and cultured,and then the expression of TF protein was detected by Western blot. The TF expression in liver tissues was measured by immunohistochemistry. The pathological damage of the pancreatic tissue was detected by HE staining. Results：The content of serum amylase was significantly higher in MAP and SAP group than in SO group（P0.01）,while there was no significant difference between MAP group,SAP group and MAP＋Gdcl3group,SAP＋Gdcl3group. The contents of TF markedly increased in MAP and SAP groups compared with those in SO group,while distinctively decreased compared with those in MAP ＋Gdcl3and SAP ＋Gdcl3group（P 0.05）. The TF protein expressions in the livers of rats in MAP and SAP groups distinctively increased compared with those in SO group. TF protein contents of KCs were obviously lower in MAP＋Gdcl3and SAP＋Gdcl3groups than in MAP and SAP groups（P0.05）. Plasma concentrations of the prothrombin time（PT）,activated partial thromboplastin time（APTT）and fibrinogen（FIB）in SAP group obviously increased compared with those in MAP groups and SO groups

关 键 词：枯否细胞 组织因子 急性胰腺炎 凝血象 

分 类 号：R657.5[医药卫生—外科学]