国际麻醉学与复苏杂志   2015, Issue (12): 2-2
    
法舒地尔对脂多糖诱导大鼠急性肺损伤的保护作用
江泽宇, 傅婕, 封光, 刘功俭1()
1.徐州医学院
The protective effect of fasudil in the lipopolysaccharide-induced acute lung injury in rats
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摘要：

目的 探讨法舒地尔（fasudil）对脂多糖（lipoplysaccharide，LPS）致急性肺损伤（ALI）大鼠的保护作用及其在可能机制。 方法 将42只雄性SD大鼠随机分为4组：对照组(C组，n=6)、LPS组(L组，n=24)、法舒地尔组(F组，n=6)、法舒地尔组+LPS(FL组，n=6)。LPS组按照不同的时间点又分为1、3、6、12小时四个亚组。L组和FL组尾静脉注射LPS 5 mg/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LPS 3h后处死大鼠取材。苏木精-伊红（hematoxylin-eosin，HE）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 weight ratio，W/D），western blot法检测Rho激酶（(Rho associated kinase ，ROCK）、紧密连接蛋白Occludin和ZO-1的表达，ELISA法检测肿瘤坏死因子（tumor necrosis factor-α，TNF-α）、白细胞介素（interleukin，IL）-6的表达。 结果 与C组相比，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-α、IL-6释放增加（P＜0.01），法舒地尔可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）；肺组织湿/干比FL组明显低于L组(4.35±0.13) vs (4.89±0.37) (P＜0.05)。 结论 在LPS引起的急性肺损伤中，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。
关键词： 急性肺损伤；LPS；法舒地尔；紧密连接蛋白；肺保护
Abstract:

Objectlve To investigate the protection of fasudil in LPS induced acute lung injury 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 acute lung injury (ALI) were developed by injection of LPS (5mg/kg) in rat tail vein in group L and group FL.1hour before injection of LPS in rat tail vein, group FL rats was injected with Fasudil (10mg/kg) through the tail vein. Group C was injected of equal volume of normal saline in rat tail vein, and group F was injected with fasudil only. 3 hours after ALI was made, all rats were killed for lung tissues collecting. The lung tissue pathology was examined by HE staining and the psychrometric ratio of lung tissue was tested by weight method. The expression of ROCK、Occludin、ZO-1 were determined by western blot．The concentrations of TNF-α and IL-6 were detected by ELISA．Results Compare with group C, the expression of ROCK were up-regulated at the time 1h, 3h, 6h, 12h after LPS injection and reach the peak at 3h after LPS injection, the expression of 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 alveol, 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-α 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＜0.05). Compared with group L, lung water content of group FL was significantly decreased (4.35±0.13) vs (4.89±0.37) (P＜0.05). Conclusion Fasudil could attenuate the injury induced by LPS in acute lung injury, the mechanism may be due to the effect of inhibiting the excessive expression of Inflammation and reducing the damage of cell junction．
Key words: Acute lung injury；LPS；fasudil；Occludin；ZO-1；Lung protection