乌司他丁对小鼠急性肺损伤保护作用及机制的研究  

Protective effect of ulinastatin on acute lung injury and its mechanism in mice

作　　者：李雪宁 曹超[1] 寿松涛[1] 王军[1] 柴艳芬[1] Li Xue-ning;Cao Chao;Shou Song-tao;Wang Jun;Chai Yan-fen(Emergency Medicine Department, Graduate College, Tianjin Medical University, Tianjin 300052, Chin)

机构地区：[1]天津医科大学总医院急诊医学科,天津300052

出　　处：《中国急救医学》2018年第6期527-532,I0002,共7页Chinese Journal of Critical Care Medicine

基　　金：天津市应用基础与前沿技术研究计划项目（13JCYBJC37500）;睿E（睿医）急诊医学研究专项基金项目（R2015026）;天普研究基金项目（UF201315）;天津医科大学总医院青年孵育基金项目（ZYYFY2015010）

摘　　要：目的 研究乌司他丁（ulinastatin, UTI）对急性肺损伤（acute lung injury，ALI）小鼠的保护作用，并探讨其机制。方法 雄性SPF级C57/BL小鼠64只，按随机数字表均分为：健康对照组（Sham组）、Sham＋UTI 组、脂多糖组（LPS组）、LPS＋UTI组。气管内滴注LPS建立ALI模型，Sham＋UTI组和LPS＋UTI组建模前30 min皮下注射UTI 10 000 U/kg。监测各组小鼠72 h生存率。建模后24 h取支气管肺泡灌洗液（BALF）和肺组织, 检测BALF中炎性细胞、中性粒细胞和巨噬细胞数；测定肺组织湿/干质量比（W/D）；观察肺组织病理学变化并计算肺损伤评分；测定肺组织中髓过氧化物酶（MPO）活性及丙二醛（MDA）含量；测定肺组织中肿瘤坏死因子-α（TNF-α）、白细胞介素-1（IL-1）、IL-6、IL-2 mRNA水平，检测肺组织中TLR4/NF-κB表达情况。结果 LPS＋UTI组小鼠生存率较LPS组明显升高，差异有统计学意义（P〈0.05）；LPS＋UTI组肺组织W/D、肺组织病理评分及肺组织中MPO活性和MDA含量较LPS组明显降低，TNF-α、IL-1、IL-6、IL-2 mRNA水平较LPS组明显降低，且TLR4/NF-κB mRNA表达水平较LPS组显著降低，差异均具有统计学意义（P〈0.05或P〈0.01）。结论 UTI可减轻LPS诱导的ALI小鼠肺组织的炎性反应，提高ALI小鼠生存率，这可能与UTI影响TLR4/NF-κB信号通路有关。Objective To investigate the protective effects and the mechanisms of ulinastatin （UTI） on LPS-induced acute lung injury （ALI） in mice. Methods Total of 64 SPF male C57/BL mice were randomly divided into four groups： Sham, Sham＋UTI, LPS, LPS＋UTI, 16 mice in each group. The mouse model of ALI was established by instilled the lipopolysaccharide （5 mg/kg） into the trachea. In Sham＋UTI group and LPS＋UTI group, mice were treated with UTI （10 000 U/kg） hypodermic injected before model establishment 30 minutes. Sham group and LPS group were treated with the equivalent normal saline. 72 h survival rate of mice were observed. Another 64 mice were divided into the above mentioned method. The bronchoalveolar lavage fluid （BALF） and lung tissue were obtained at 24 h after LPS treatment. The total cell count, neutrophil count and macrophage count in the BALF were determined; the wet/dry （W/D） weight ratio of the lung tissue were measured; hispathological changes and pathology scores of the lung tissue were observed by HE staining; myeloperoxidase （MPO） activity and malondialdehyde （MDA） contents in the lung tissue homogenate were measured; mRNA level of tumor necrosis factor （TNF-α）, interleukin-1 （IL-1）, interleukin-6（IL-6）, interleukin-2 （IL-2） in the lung tissue were measured by real-time PCR; the expression of TLR4/NF-κB p-65 were detected by RT-PCR and Western blot. Data analysis using SPSS 21.0 Software, multiple-group comparisons were performed using one-way ANOVA, the intergroup comparisons were analyzed by the LSD-t test with P〈0.05 for the statistically significant difference. Results The survival rate, lung W/D ratio, hispathological changes, pathological scores, pulmonary MPO activities and MDA contents, mRNA levels of TNF-α, IL-1, IL-6, IL-2, TLR4/NF-κB mRNA were all lower in LPS＋UTI group than those in LPS group, the difference was statistically significant （P〈0.05 or P〈0.01）. Conclusion UTI can inhibit the inflammatory respons

关 键 词：乌司他丁(UTI) 小鼠 脂多糖(LPS) 急性肺损伤(ALI) Toll样受体4(TLR4) NF-κB信号通路 机制 

分 类 号：R818.74[医药卫生—放射医学]