乌司他丁对烟雾吸入性肺损伤大鼠肺组织细胞凋亡和炎症的影响  

Effects of ulinastatin on apoptosis and inflammation of lung tiss ue in rats with smoke inhalation injury

作　　者：刘欣健 崔正军[1] 张树堂[1] 王常印[1] 郭鹏飞[1] 孟庆楠[1] 周健[1] 邹仕波[1] 王旭[1] 余祖改 Liu Xinjian;Cui Zhengjun;Zhang Shutang;Wang Changyin;Guo Pengfei;Meng Qingnan;Zhou Jian;Zou Shibo;Wang Xu;Yu Zugai(Department of Burn Repair and Reconstruction Surgery,First Hospital Affiliated to Zhengzhou University,Zhengzhou Henan,450000,China)

机构地区：[1]郑州大学第一附属医院烧伤修复重建外科,河南郑州450000

出　　处：《感染．炎症．修复》2020年第2期88-91,F0002,共5页Infection Inflammation Repair

基　　金：国家自然科学基金资助项目(U1604188)。

摘　　要：目的:探讨乌司他丁对烟雾吸入性肺损伤大鼠肺组织细胞凋亡及白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)含量的影响。方法:SD大鼠27只,体重200 g,随机分为对照组、吸入组和乌司他丁组。乌司他丁组和吸入组大鼠建立烟雾吸入性损伤模型,乌司他丁组大鼠于模型制备成功后经腹腔注射乌司他丁1×104 U/kg,对照组和吸入组以等量生理盐水代替乌司他丁。24 h后取血和肺组织标本,酶联免疫吸附法检测血清IL-6和TNF-α含量;TUNEL法检测肺组织细胞凋亡,计算细胞凋亡指数;免疫组化法测定肺组织Bax和Bcl-2蛋白表达,Western blot法检测肺组织蛋白提取物中IL-6、cleaved-caspase-3、TNF-α的蛋白表达情况。结果:对照组肺泡腔清晰、完整,间质无炎细胞浸润、肿胀,吸入组肺泡水肿、充血,炎细胞浸润,乌司他丁组较吸入组炎症明显减轻。吸入组凋亡指数明显高于对照组(P<0.05),乌司他丁组较吸入组凋亡指数明显减少(P<0.05)。免疫组化结果显示:促凋亡蛋白Bax在吸入组表达较多,在对照组和乌司他丁组表达相对较少;抑凋亡蛋白Bcl-2在吸入组表达较少,在对照组和乌司他丁组表达较多。Western blot结果显示,吸入组肺组织IL-6、TNF-α、cleaved-caspase-3的蛋白表达明显多于对照组(P<0.05),乌司他丁组的IL-6、cleaved-caspase-3、TNF-α蛋白表达明显少于吸入组(P<0.05)。结论:乌司他丁能抑制烟雾吸入导致的肺组织细胞的凋亡及炎症反应。Objective:To investigate the effect of ulinastatin on apoptosis and inflammation of lung tissue in rats with smoke inhalation injury.Methods:Twenty-seven SD rats weighting 200 g were randomly divided into three groups:control(Cont)group,inhalation(Inh)group and ulinastatin(Uli)group.Animal models of smoke inhalation injury were established.Rats in Uli group were intraperitoneally injected with 10000 U/kg of ulinastatin after successful modeling,and the same amount of normal saline was injected intraperitoneally in Cont group and Inh group.Twentyfour hours later,the abdominal aorta blood were sampled and the upper sera were collected to determine the serum levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)by ELISA.Lung tissue specimens were retained 24 hours post-treatment to detect the apoptosis of lung tissue by TUNEL method,the expressions of Bax and Bcl-2 protein by immunohistochemistry,and the expressions of IL-6,cleaved-caspase-3 and TNF-αprotein by Western blotting.Results:Histopathologic study showed full and distinct alveolar space in lung tissue of the rats in Cont group,with no swelling and invasion of inflammatory cells in interstitial tissues.Obviously swelled and congestive alveolus as well as infiltration of inflammatory cells were observed in the Inh group,while the inflammation was relieved markedly in Uli group than in Inh group.The apoptosis index was significantly higher in Inh group than in Cont group(P<0.05),and the index was obviously lower in Uli group than in Inh group(P<0.05).The expression of the pre-apoptotic protein Bax was higher in Inh group,but relatively lower in Cont group and Uli group;while the expression of anti-apoptotic protein Bcl-2 was just the reverse.The results of Western blotting showed that the expressions of IL-6,TNF-αand cleavedcaspase-3 protein in the lung tissue were significantly higher in Inh group than in Cont group(P<0.05),while they were significantly lower in Uli group than in Inh group(P<0.05).Conclusions:Ulinastatin can inhibit the apoptosi

关 键 词：乌司他丁 肺损伤 吸入性 细胞凋亡 炎症 

分 类 号：R64[医药卫生—外科学]