法舒地尔对脂多糖诱导大鼠急性肺损伤的保护作用  

The protective effect of fasudil in the lipopolysaccharide-induced acute lung injury in rats

作　　者：江泽宇 傅婕[1] 封光[2] 焦皓[2] 刘功俭[2] 

机构地区：[1]徐州医学院江苏省麻醉学重点实验室、江苏省麻醉与镇痛应用技术重点实验室,221004 [2]徐州医学院附属医院麻醉科

出　　处：《国际麻醉学与复苏杂志》2015年第12期1062-1066,1085,共6页International Journal of Anesthesiology and Resuscitation

基　　金：国家自然科学基金（81200056）

摘　　要：目的探讨法舒地尔对脂多糖（lipoplysaccharide，LPS）致急性肺损伤（acutelunginjury，ALI）大鼠的保护作用及其可能机制。方法将42只雄性SD大鼠按随机数字表法分为4组：对照组（C组，6只）、LPS组（L组，24只）、法舒地尔组（F组，6只）、法舒地尔＋LPS组（FL组，6只。LPS组按照不同的时间点又分为1、3、6、12h4个亚组。L组和FL组尾静脉注射LPS5mg／kg制作内毒素性肺损伤模型，FL组尾静脉注射LPS前1h尾静脉注射法舒地尔（10mg／kg），C组尾静脉注射等量生理盐水，F组尾静脉注射等量法舒地尔。注射LPS3h后处死大鼠取材。苏木精伊红（hematoxylin-eosin，liE）染色法检查肺组织病理变化，称重法检测肺组织的湿／干重比（wet／dry，W／D），Western blot法检测Rho激酶（Rho associated kinase，ROCK）、紧密连接（tight junction，TJ）蛋白中闭锁蛋白（Occludin）和闭锁小带蛋白1（zonula occludens 1，ZO-1）的表达，酶联免疫吸附实验（enzyme linked immunoserbent assay，ELISA）法检测肿瘤坏死因子啦（tumor necrosis factor-a，TNF-a）、白细胞介素（interleukin，IL）-6的表达。结果与C组比较，注射LPS后1、3、6、12h肺组织中ROCK蛋白的表达明显增加，并于3h达到最大值，ZO-1和Occludin蛋白的表达明显降低，LPS诱导的ALI大鼠肺组织结构破坏明显，可见肺泡内出血较多，肺泡委陷，伴有大量炎性细胞浸润，间质明显水肿，肺泡灌洗液（bronchoalveolar lavage fluid，BALF）中TNF-a、IL-6释放增加（P〈0．01）。法舒地尔町以部分逆转上述病理改变（P〈0．05），且法舒地尔可以增加ALI大鼠肺组织内Occludin、ZO-1蛋白的表达（P〈0．05）；FL组肺组织W／D明显低于L组[（4．35±0．13）比（4．89±0．37）]（P〈0．05）。结论在LPS引起的ALI中，法舒地尔可以通过抑制过度表达的炎症反应、减轻细胞连接的破坏从而产生肺保护作用。Objective To investigate the protection of fasudil in lipopolysaccharide （LPS） induced acute lung injury （ALl） in rats and its possible mechanism. Methods Forty two male SD rats were randomly divided into four groups： control group （group C, n=6）, LPS group （group L, n=24）, fasudil group （group F, n=6）, LPS＋fasudil group （group FL, n=6）. LPS group was further divided into four subgroups. The rat models of ALI were developed by injection of LPS （5 mg/kg） in rat tail vein in group L and group FL. One hour before injection of LPS in rat tail vein, group FL rats was injected with fasudil （ 10 mg/kg） through tile tail vein. Group C was injected of equal volume of normal saline in rat tail vein, and group F was injected with fasudil only. Three hours after ALI was made, all rats were killed for lung tissues collecting. The lung tissue pathology was examined by hematoxylin-eosin （HE） staining and the psychrometric ratio of lung tissue was tested by weight method. The expression of Rho associated kinase （ROCK）, Occludin, ZO-1 were determined by Western blot. The concentrations of tumor necrosis factor-a （TNF-a） and interleukin （IL）-6 were detected by enzyme linked immunoserbent assay（ELISA）. Results Compare with group C, the expression of ROCK were up- regulated at the ＂time 1, 3, 6, 12 h after LPS injection and reach the peak at 3 h after LPS injection, the expression of zonula occludens 1 （ZO-1） and Occludin were down-regulated significantly. The lung tissues of rat models of ALI induced by LPS showed obviously destruction, excessive bleeding in pulmonary alveoli, and inflammatory responses under light microscopy, characterized by alveolar hemorrhage, pulmonary interstitial edema, alveolar collapse and massive inflammatory cell infiltration within the lung tissues, LPS up-regulated the expression of TNF-a and IL-6 in BALF （P〈0.01）. Fasudil could partly reversed the above role （P〈0.05）, and also induced the up-regulation of Occludin and ZO-1 （P�

关 键 词：脂多糖 急性肺损伤 法舒地尔 肺保护 

分 类 号：R563.8[医药卫生—呼吸系统]