抗凝血酶Ⅲ对油酸诱导急性肺损伤大鼠的肺保护作用  

Effects of Antithrombin-Ⅲ on Inflammatory Reaction and Pulmonary Protection in Oleic Acid-Induced Acute Lung Injury Rats

作　　者：孙辉明[1] 施毅[1] 宋勇[1] 高伟[1] 李培[1] 孙文逵[1] 

机构地区：[1]南京大学医学院南京军区南京总医院呼吸内科,江苏南京210002

出　　处：《中国呼吸与危重监护杂志》2011年第2期148-153,共6页Chinese Journal of Respiratory and Critical Care Medicine

基　　金：江苏省自然科学基金(编号:BK2009318);南京军区医学重点课题基金(编号:09Z028);南京军区南京总医院重点课题基金(编号:Z2008014)

摘　　要：目的观察抗凝血酶Ⅲ对油酸诱导急性肺损伤大鼠的肺保护作用,并探讨相关机制。方法 60只清洁级雄性SD大鼠按随机数字表法分为正常对照组、急性肺损伤组、抗凝血酶Ⅲ治疗组、抗凝血酶Ⅲ联合肝素治疗组和肝素治疗组。油酸(0.2 mL/kg)静脉注射建立急性肺损伤大鼠模型。改良Smith肺损伤病理评分法评价肺损伤程度,发色底物法测定血浆中抗凝血酶Ⅲ的活性,比重法测定肺组织血管外肺水(EVLW)和肺组织湿/干重比(W/D),单核素示踪技术测定肺微血管白蛋白通透性(Palb),酶联免疫吸附法(ELISA)测定血清中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)和血管假性血友病因子(vWF)含量,蛋白质印迹法检测肺组织细胞外信号调节激酶(ERK)1/2、P38丝裂原活化蛋白激酶(P38 MAPK)和c-jun氨基端激酶(JNK)磷酸化蛋白表达。结果 (1)正常对照组Smith肺损伤病理评分为(0.67±0.52)分,显著低于急性肺损伤组(11.76±2.23)、抗凝血酶Ⅲ治疗组(10.98±3.42)、抗凝血酶Ⅲ联合肝素治疗组(12.07±1.83)和肝素治疗组(12.54±3.78)(P均<0.01)。急性肺损伤组Smith肺损伤病理评分与各治疗组比较差异无统计学意义(P>0.05)。(2)各组间血浆抗凝血酶Ⅲ的活性差异无统计学意义(P均>0.05)。(3)急性肺损伤组Palb为0.50±0.07,显著高于正常对照组(0.20±0.02,P<0.01),与抗凝血酶Ⅲ治疗组(0.47±0.12)、抗凝血酶Ⅲ联合肝素治疗组(0.46±0.08)和肝素治疗组(0.48±0.06)相比,差异均无统计学意义(P>均0.05)。(4)急性肺损伤组EVLW为(1.14±0.12)mL/kg,显著高于正常对照组[(0.69±0.04)mL/kg,P<0.01],与抗凝血酶Ⅲ治疗组[(1.12±0.21)mL/kg]、抗凝血酶Ⅲ联合肝素治疗组[(1.08±0.13)mL/kg]和肝素治疗组[(1.14±0.20)mL/kg]相比,差异均无统计学意义(P均>0.05)。(5)急性肺损伤组TNF-α和IL-6的含量分别为(1.613±0.238)μg/L和(0.685±0.129)ng/mL,显著高于正常对照组[(0.506±0.093)μg/L和(0.233±0.047)ng/mL,P�Objective To investigate the effects of antithrombin-Ⅲ(AT-Ⅲ) on the inflammatory reaction in oleic acid-induced acute lung injury(ALI) rats.Methods Sixty male Sprague-Dawley rats were randomly divided into five groups,ie.a normal control group,an ALI group,an AT-Ⅲ treatment group,an AT-Ⅲ+heparin treatment group,and a heparin treatment group(n=12).The ALI rats were induced by injecting oleic acid(0.2 mL/kg) intravenously.The lung histology was scored by modified Smith technique.The albumin permeability of pulmonary microvascular(Palb) was measured by single nuclide tracer technique.The extravascular lung water(EVLW) and wet/dry weight ratio(W/D) of lung tissues were measured by gravity way.The activity of AT-Ⅲ in plasma was determined by the method of synthetic chromogenic substrate.Tumor necrosis factor α(TNF-α),interleukin 6(IL-6) and von Willebrand factor(vWF) levels in serum were determined using commercial enzyme-linked immunosorbent assay kits.The expressions of lung tissue extacellular signal-regulated kinases(ERK)-1/2,P38 mitogen-activated protein kinase(MAPK) and c-jun N-terminal kinases(JNK) were determined by Western blot.Results The Smith scores,EVLW,Palb,plasma level of vWF,lung tissue levels of phospho-ERK1/2 and phospho-P38 MAPK expressions in the ALI group were all significantly higher than those in the normal control group(P<0.05),while not significant different with other three treatment groups.There were not significant differences in the activity of AT-Ⅲ in plasma and phospho-JNK expression among all five groups.The serum levels of TNF-α and IL-6 in the ALI group were significantly higher than those in the normal control group and three treatment groups.Conclusions AT-Ⅲ downregulates the levels of downstream cytokines TNF-α and IL-6,but can not inhibite the activation of ERK1/2 and P38 MAPK,and can not relieve endothelial permeability.The study do not demonstrate the lung protective effect of AT-Ⅲ in oleic acid-induced acute lung injury.

关 键 词：急性肺损伤 抗凝血酶Ⅲ 油酸 丝裂原活化蛋白激酶 

分 类 号：R965[医药卫生—药理学]