乌司他丁对脓毒症大鼠肺损伤抑炎及抗凋亡信号机制的研究  

The protective effect and ameliorating cell apoptosis of Ulinastatin on lung tissue in septic rats

作　　者：李永宁[1] 王磊[1] 陈俊杰[1] 李雪娇[1] 王立峰[1] 康健[1] Li Yong-ning;Wang Lei;Chen Jun-fie;Li Xue-jiao;Wang Li-feng;Kang Jian(Department of Emergency,the First Affiliated Hospital of Medical University,Dalian 116011,China)

机构地区：[1]大连医科大学附属第一医院急诊科,辽宁大连116001

出　　处：《中国急救医学》2018年第7期626-630,I0004,共6页Chinese Journal of Critical Care Medicine

基　　金：辽宁省自然科学基金项目（201403858）;大连市医学科学研究计划项目（1812007）

摘　　要：目的 探讨乌司他丁（UTI）对脓毒症（sepsis）大鼠肺脏保护作用及其抗凋亡机制的研究。方法 SD雄性大鼠随机分三组：Sham组（n=10）仅行开关腹手术，Sepsis组（n=10）和UTI组（n=10）行盲肠结扎穿孔术（CLP），UTI组建模前1 h大鼠尾静脉注射乌司他丁20万U/kg，24 h后重复给药，Sepsis组注射等量生理盐水。于6、12、24、36、48 h尾静脉采血测定TNF-α、IL-6、IL-10浓度，48 h留取肺脏标本行苏木素-伊红（HE）染色和免疫组化检测Bax/Bcl-2、TUNEL表达及Western-blot检测，caspase-3蛋白表达，透射电镜下观察肺组织超微结构变化。结果 UTI组炎症因子TNF-α、IL-6表达高于Sham组而低于Sepsis组（P〈0.05），UTI组抑炎因子IL-10高于Sepsis组而低于Sham组（P〈0.05）。HE染色Sepsis组肺泡水肿、充血，炎细胞侵润，而UTI组较Sepsis组明显减轻；在Sepsis组凋亡蛋白Bax表达高于UTI组，而抗凋亡蛋白Bcl-2表达低于UTI组；TUNEL检测肺泡上皮细胞凋亡率UTI组明显低于Sepsis组（P〈0.05）；透射电镜观察，Sepsis组肺泡壁及毛细血管基底膜肿胀、肺泡腔变小，腔内大量炎性渗出、肺泡Ⅱ型上皮细胞内线粒体空化等，而UTI组明显减轻；通过Western-blot检测，caspase-3蛋白在UTI组表达高于Sham组而低于Sepsis组（P〈0.05）。结论 乌司他丁抑制脓毒症大鼠血清中促炎介质的释放，抗血管内皮细胞及肺泡上皮细胞凋亡，降低caspase-3蛋白表达，对脓毒症大鼠急性肺损伤（ALI）起到保护作用。Objective To investigate the protective role of Ulinastatin on lungs of rats with sepsis and its anti -apoptosis mechanism. Methods 30 male SD rats were randomly divided into three groups ： the Sham group simply underwent open - close surgery; both the sepsis group and UTI group underwent cecal ligation and puncture （CLP） ; in UTI group, Ulinastatin 200,000U/kg was given 1 h prior to modeling, and was repeatedly given at 24 h post - treatment; the sepsis group received injection of normal saline. Caudal venous blood was collected at 6, 12, 24, 36, 48 h post - treatment to determine the mass concentrations of TNF-α, IL- 6, and IL- 10; lung specimens were retained at 48 h post - treatment to detect the expression of Bax/Bcl - 2 and TUNEL by HE staining and IHC, and the expression of caspase - 3 by Western - blot ; uhrastructural changes in lung tissues were observed under transmission electron microscope （TEM）. Results In UTI group, expression of the inflammatory cytokines TNF - α and IL - 6 was higher than that in Sham group and lower than that in sepsis group （ P 〈 0.05 ） ; and the expression of the anti - inflammatory cytokine IL - 10 was higher than that in sepsis group and lower than that in Sham group （ P 〈 0.05 ）. HE staining revealed alveolar edema, congestion and inflammatory cell infiltration in sepsis group, which were significantly alleviated in UTI group ; in sepsis group, expression of the apoptotic protein Bax was higher than that in UTI group, while expression of the anti - apoptotic protein Bcl - 2 was lower than that in UTI group ; the alveolar epithelial cell apoptosis rate detected by TUNEL was significantly lower in UTI group than in sepsis group （P 〈 0.05）; alveolar wall and capillary basement membrane swelling, reduced alveolar space, massive inflammatory exudation in alveolar space, and mitochondrial cavitation in type II penumonoeytes were observed under TEM, which were significantly alleviated in UTI group; the expression of caspase - 3 protein detected by Wes

关 键 词：乌司他丁 脓毒症 炎症 凋亡 Bax Bcl-2 Caspase-3 原位缺口末端标记法(TUNEL) 急性肺损伤(ALI) 

分 类 号：R589.2[医药卫生—内分泌]