摘要
目的 :研究神经激肽 1受体 (NK 1R)在急性坏死性胰腺炎 (ANP)大鼠肺组织中的表达 ,探讨该受体在 ANP肺损害中的作用。方法 :健康成年 Sprague Dawley大鼠按抽签法随机分为 ANP组 (90只 )和正常对照组 (30只 )。正常对照组开腹后只翻动胰腺 ,ANP组大鼠经胰胆管恒速逆行注射质量分数为 5 %的牛磺胆酸钠 (0 .1ml/ kg)制成 ANP大鼠模型。检测肺脏髓过氧化物酶 (MPO)的活性和肺脏毛细血管通透性 (L CP)。应用逆转录聚合酶链反应 (RT PCR)检测肺组织中 NK 1R m RNA水平 ,应用 Western Blot技术检测NK 1R的蛋白水平 ,以免疫组织化学方法进行 NK 1R的组织学定位。结果 :ANP组 6 h后肺组织 MPO和L CP水平即明显高于正常对照组。与正常对照组相比 ,ANP肺组织中 NK 1R m RNA和蛋白都过度表达 ;NK 1R m RNA表达分别与 MPO(r=0 .83,P<0 .0 1)和 L CP(r=0 .79,P<0 .0 1)水平相关。免疫组织化学检测显示 ,NK 1R的表达主要位于肺泡隔血管内皮细胞表面及部分肺泡 型、 型上皮细胞表面等处。结论 :ANP肺组织中 NK 1R的表达水平明显上调 ,导致中性粒细胞等炎性细胞聚集 ,加剧 ANP时的肺损害。
Objective: To investigate the expression of neurokinin1 receptor (NK1R) in the lung tissue, and the relationship between expression of NK1R and lung injury in rats with acute necrotizing pancreatitis (ANP). Methods: One hundred and twenty adult SpragueDawley rats were randomly divided into ANP and control groups. Animals in group ANP were induced by the retrograde intraductal infusion of 5% sodium taurocholate (0 1 ml/kg),and animals in normal control group received laparotomy only. The accumulation of polymorphonuclear leukocytes in lung tissues was measured with myeloperoxidase (MPO) assay. Lung endothelial barrier destruction was measured by lung capillary permeability (LCP). Reverse transcription polymerase chain reaction (RTPCR) was used to determine the mRNA expression of NK1R, western blot analysis was used to determine NK1R protein expression levels, and immunohistochemistry was used to localize expression site of NK1R. Results: NK1R mRNA level was enhanced in the lung of ANP compared with normal control group. Western blot analysis showed overexpression of NK1R protein level exited in ANP group. Statistical analysis revealed correlation between NK1R mRNA and MPO ( r =0 83, P <0 01) and LCP ( r =0 79, P <0 01) respectively. With immunohistochemistry staining, moderate to strong NK1R immunoreactivity was localized to alveolar membrane, Ⅰ epithelium, Ⅱepithelium and polymorphonuclear leukocytes in the lung of ANP. Conclusion: In ANP, overexpression of NK1R contributes to disturbance of neuropeptides loop, resulting in aggregation of neutrophilic granulocyte and promoting deterioration of lung injury.
出处
《中国危重病急救医学》
CAS
CSCD
2003年第7期422-425,共4页
Chinese Critical Care Medicine
基金
国家人事部留学回国人员科研启动基金资助项目( 7690 0 0 40 2 7)
