抑制小窝蛋白-1磷酸化对机械通气所致肺损伤的保护作用  

Protection of the ventilation - induced lung injury by inhibiting Caveolin - 1 phosphorylation

作　　者：戴春光[1] 肖军[1] 钟荣[1] 于志辉[1] 周吉[1] 王文艳[1] 

机构地区：[1]桂林医学院附属医院重症医学科,广西桂林541001

出　　处：《中国急救医学》2015年第8期735-740,I0002,共7页Chinese Journal of Critical Care Medicine

基　　金：广西壮族自治区自然科学基金（2010GXNSFA013253）

摘　　要：目的：研究抑制小窝蛋白－1（ Cav－1）磷酸化对机械通气所致肺损伤的效应。方法50只健康雄性SD大鼠随机分为A、B1、B2、C1及C2五组（每组10只）。 A组为正常对照组，仅做气管切开；B1组为1 h大潮气量机械通气组；B2组为2 h大潮气量机械通气组；C1组为1 h大潮气量机械通气＋酪氨酸蛋白激酶抑制剂PP2（5 mg／kg）组；C2组为2 h大潮气量机械通气＋酪氨酸蛋白激酶抑制剂PP2（5 mg／kg）组。 C1和C2组在机械通气前1 h腹腔注射抑制剂。各组大鼠均连续监测平均动脉压（ MAP ）。光镜观察肺组织病理改变，并做弥漫性肺泡损伤系统（ diffuse alveolar damage， DAD）评分，比色法测定髓过氧化物酶（ myeloperoxidase， MPO）活性，计算肺湿／干比（W／D），蛋白印迹检测磷酸化小窝蛋白－1[P －Cav －1（Y14）]和小窝蛋白－1（Cav－1）表达情况，免疫组织化学染色法检测肺组织中内皮型一氧化氮合酶（eNOS）和白细胞介素－1受体相关激酶4（IRAK4）表达情况，酶联免疫吸附试验（ELISA）检测各组大鼠肺泡灌洗液TNF－α水平，伊文思蓝（ Evans blue）法检测肺血管通透性。结果 B1、B2、C1、C2组病理损伤评分、肺湿／干比、Cav－1、eNOS、IRAK4表达量、TNF－α水平、Evans blue渗出量、MPO活性、MAP及血气分析与A组比较差异均有统计学意义（均P＜0．05）；与A组比较，B1、B2、C1组P－Cav－1表达升高（均P＜0.05），而C2组P－Cav－1（Y14）表达量差异无统计学意义（P＞0．05）；C1组与B1组比较，C2组与B2组比较，病理损伤评分、肺湿／干比，P－Cav－1（Y14）、eNOS、IRAK4表达量、TNF－α的水平、Evans blue含量、MPO活性、MAP、及血气分析差异均有统计学意义（均P＜0.05）；C1组与B1组比较，C2组与B2组比较，Cav－1表达量差异无统计学意义（均P＞0．05）。结论抑制Cav－1磷酸化能够降低肺毛细血管通透�Objective To define the protective role of Caveolin-1 phosphorylation in the lung injury induced by ventilation.Methods Fifty healthy male SD rats were randomly divided into five groups：A, B1, B2, C1, C2 groups （n=10）.Group A served as control group, in which rats did not receive ventilation.Groups B1, B2, C1 and C2 received large-tide volume ventilation, but only rats in groups C1 and C2 received tyrosine kinase inhibitor 4-amino-5-（4-chlorophenyl）-7-（t-butyl） pyrazolo[3,4-d] pyrimidine （PP2, 5 mg/kg respective） 1 hour before ventilation.After maintaining ventilation for 1 h or 2 h, all rats were sacrificed and specimens of lung tissues and bronchoalveolar lavage fluid （ BALF） were harvested.Mean arterial pressure （ MAP） was observed continuously.Lung pathological changes were observed by microscopy and diffuse alveolar damage （ DAD ） score were estimated.The protein levels of eNOS and interleukin-1 receptor-associated kinase 4 （ IRAK4 ） in lung tissues were determined by immunohistochemistry staining.The expression levels of P -Cav -1 （ Y14） and Cav-1 in lung tissue were measured by Western Blotting.Lung myeloperoxidase （ MPO） activity was measured by colorimetric analysis, wet-dry weight ratio （ W/D） was measured.The levels of tumor necrosis factor -α（ TNF -α） in BALF were measured by enzyme -linked immunosorbent assay （ ELISA ） .Evans Blue was used to detect the pulmonary vascular permeability.Arterial blood gases were measured every one hour. Results Comparison with group A, there was statistical significance in the expression of Cav-1, eNOS and IRAK4, protein in lung tissue, W/D ratio, the amount of Evans blue extravasation, diffuse alveolar damage （ DAD） score, MAP, arterial blood gas analyses, as well as the amount of MPO and TNF-αin BALF among group A, group B1, group B2, group C1 and group C2 （all P〈0.05）.Comparison with group A there was statistical significance in the expression of P -Cav -1 among the five groups but group C2 （P ＆g

关 键 词：小窝蛋白-1(Cav-1) 磷酸化小窝蛋白-1 酪氨酸蛋白激酶抑制剂 机械通气 机械通气所致肺损伤 

分 类 号：R-332[医药卫生]