已酮可可碱治疗急性胰腺炎合并急性肺损伤的实验研究.doc

已酮可可碱治疗急性胰腺炎合并急性肺损伤的实验研究已酮可可碱治疗急性胰腺炎合并急性肺损伤的实验研究更新日期：2010-07-03 何新华周玉淑邵孝洪黄席珍王仲赵向一于学忠 【摘要】目的探讨急性肺损伤的治疗方法。方法建立兔急性肺损伤模型,随机分3组,筛选药物已酮可可碱治疗,检测动脉血气,血清TNF-含量和活性,支气管肺泡灌洗液计数白细胞和白蛋白,肺血管通透性以及肺组织大体和病理观察。结果实验组与模型组比较,血气改善,血清TNF-的含量和活性明显降低,白细胞计数下降,白蛋白含量减低,肺血管通透性下降,肺组织大体明显改善,病理观察损伤肺组织修复;与对照组差异不明显。结论已酮可可碱能有效改善急性胰腺炎合并急性肺损伤。 【关键词】已酮可可碱急性胰腺炎急性肺损伤 Experimental Study of Pentoxifylline in Acute Pancreatitis Combined with Acute Lung InjuryHE Xin-hua, ZHOU Yu-shu, SHAO Xiao-hong, et al. Dept. of Emergency, General Hospital of Beijing Military Command, Beijing 100700 【Abstract】AimTo explore a therapeutic method for experimental acute pancreatitis (EAP) combined with acute lung injury (ALI). MethodsRabbit models of acute pancreatitis combined with acute lung injury were randomly divided into the model group (group A), the pentoxifylline group (group B) and the saline solution group (group C). The arterial blood gas and serum TNF- content and activity were analyzed. The neutrophile and polymorphnuclear leukocyte (PMN) in the bronchoalveolar lavage fluid (BALF) and macrophage (MA) were also observed respectively. The albumin content and lung extration were analyzed with the pulmonary microvascular permeability index (PMPI). The surface and histological appearence of lung were observed under microscope and electromicroscope. ResultsThere were more obvious changes in arterial gas, TNF- content and activity, PMN, MA, albumin content, PMPI and the surface and histological appearence of lung in group B than that in group A, and there were no obvious difference in group B and in group C. ConclusionIt suggests pentoxifylline can significantly improve the ALI combined with EAP in rabbits. 【Key words】Pentoxifylline Acute pancreatitisAcute lung injury 急性肺损伤可由多种原因引起。菌血症、误吸高pH胃肠液、多发创伤、急性胰腺炎、自身免疫疾病介导的器官损害、缺血再灌注等都可成为诱因。细菌感染时,中性粒细胞趋化、吞噬、产生氧自由基,释放蛋白酶以及脱颗粒,吞噬和杀伤侵入的病原微生物,保护组织免受损害和感染。当中性粒细胞过度激活,则表现出破坏作用,导致肺损伤。 已酮可可碱(PTX)因能改善血液流变学而用于治疗血管功能不全性疾病1。近年来发现它能抑制中性粒细胞聚集激活,用于治疗炎症性疾病,取得一定疗效2。笔者旨在探讨PTX对中性粒细胞和TNF-的作用过程,进而了解治疗急性肺损伤的可行性及机制。 材料与方法 1.材料:实验兔由中国医学科学院动物所提供,6个月龄、重量(2.180.31)kg,雄性,总计48只。 2.实验步骤:随机分为3组,每组16只。每组取半数动物行肺微血管通透性检查。动物实验前14小时禁食,8小时禁水。模型制备:3%戊巴比妥钠(30mg/kg)腹腔注射,麻醉成功后固定动物开腹,显露十二指肠。于十二指肠乳头部上下各1.5cm处环扎,形成闭合肠袢。此前肠腔内埋入长约4cm、内径4mm塑料导管,以利胃肠液通畅流动,同时留置股动静脉插管。A组:闭合肠袢内注入5%牛磺胆酸溶液和5%胰蛋白酶溶液各0.5ml/kg。静脉注射等渗盐水0.5ml.kg1.h1,泵入维持血压,动脉插管监测血压。B组:闭合肠袢注入药物同实验组,在完成模型复制前1小时,静脉缓慢推入1%PTX 10mg/kg,密切观察血压变化,在血压波动不明显的情况下继续5mg.kg1.h1 1%PTX静脉泵入,维持至实验结束。C组:闭合肠袢注入相同体积的等渗盐水,其他同A组。 3.检测指标及方法:(1)0,2,4,6小时取动脉血测动脉血氧分压。(2)TNF-含量测定:按照TNF-试剂盒进行操作(TNF-试剂盒由北京邦定生物医学公司提供)。(3)TNF-活性测定:运用L-929细胞法3测TNF-活性。(4)支气管肺泡灌洗液(BALF):白细胞计数、分类及白蛋白含量测定:6小时(实验结束)后活杀动物,取右肺组织,从右肺支气管置入一塑料导管,缓慢注入10ml等渗盐水,轻轻震动肺组织,5分钟后回收灌洗液至恒量。记录回收量,1000r/min离心,沉渣制成悬浮液后作白细胞计数并分类。BALF用溴甲酚绿法测白蛋白含量。(5)肺血管通透性测定(PMPI):实验第5小时经股静脉注射1%Evans Blue(2ml/kg),1小时后经股动脉放血处死。开胸,剪开左心房,将连接输液装置的9号针头插入肺动脉圆锥或右心房,在100cmH2O(9.8kPa)的压力下用等渗盐水灌洗肺血管,至左心房无血性液体流出为止。剪下右肺组织约1g,称重后置洁净小瓶中,按3ml/kg湿肺组织加入99%甲酰胺,37孵育24小时。浸出液于623nm波长处测定吸光度。PMPI以Evans Blue漏出量g/g干