Wnt5a与Syndecan-1在小鼠脓毒症致急性肺损伤中的表达研究  

The expression and significance of Wnt5a and Syndecan-1 in sepsis-induced acute lung injury of mice

作　　者：齐颖[1] 李真玉[1] 王斌[1] 陈兵[1] Qi Ying Li Zhen-yu Wang Bin Chen Bing(Intensive Care Unit, the Second Hospital of Tianjin Medical University, Tianjin 300211, China)

机构地区：[1]天津医科大学第二医院ICU,天津300211

出　　处：《中国急救医学》2017年第1期43-48,F0003,共7页Chinese Journal of Critical Care Medicine

基　　金：天津市卫生和计划生育委员会科技基金项目（2014KZ103）

摘　　要：目的研究Wnt5a及Syndecan-1的表达在脓毒症致急性肺损伤（ALI）模型中的作用。方法制备脓毒症致ALI动物模型，小鼠随机分为ALI组和对照组。小鼠按时间点分为1、3、5、8、12、24h，每组8只，研究血清Wnt5a及Syndecan-1浓度、血管通透性及肺组织病理的改变。ALI组腹腔注射大肠杆菌脂多糖（lipopolysaccharide，LPS），对照组注射等体积生理盐水，取血保留上清备用，采用双抗体一步夹心法酶联免疫吸附试验（EusA）测定血清Wm5a及Syndecan-1的浓度；小鼠左肺组织进行HE染色，右肺上叶进行匀浆提取蛋白，Westernblot检测Wnt5a蛋白的表达；右肺下叶进行组织mRNA的提取，并进行逆转录及Real—timePCR观察Syndecan-1基因的表达水平；尾静脉注射伊文思蓝（Evansblue，EB）观察肺血管渗透性；透射电镜下观察右中叶肺组织的血管内皮细胞结构改变。结果ALI组的肺组织肺泡壁增厚，血细胞及炎性细胞大量渗出；随着肺损伤程度加重，血清Wnt5a及Syndecan-1浓度逐渐升高，分别在5h及3h达峰值，在24h内维持较高浓度；Wnt5a蛋白表达水平随肺损伤程度加重而表达增加，Syndecan-1 mRNA在3h开始表达上调；甲酰胺中伊文思蓝的浓度随肺损伤程度加重逐渐升高，与对照组比较，电镜下观察到肺血管内皮细胞肿胀显著，细胞内空泡增加，部分细胞内线粒体肿胀，细胞间距增宽，基膜与内皮细胞分离。结论肺脏血管内皮损伤发生在Au早期，血清WntSa与Syndecan-1浓度改变与肺损伤严重程度一致，二者作为ALI的早期生物学标志物为临床液体管理提供一定指导作用。Objective To evaluate the induction of Wnt5a and Syndecan-1 in sepsis-induced acute lung injury （ALI） of mice. Methods We established the animal model of sepsis-induced acute lung injury. Mice were randomly divided into two groups as follows： ALI group and control group. The mice in each group were divided into 6 subgroups according to the time of sacrifice： 1 h, 3 h, 5 h, 8 h, 12 h, 24 h, with 8 mice in each subgroup. We detected the concentration of Wnt5a and Syndecan-1 in serum, lung vascular permeability and the pathological changes of lung tissue. ALI group was injected Escherichia coli lipopolysaccharide （LPS） by intraperitoneal injection, and the control group was injected with an equal volume of saline. Serum Wnt5a and Syndecan- 1 were determined by double antibody sandwich enzyme linked immunosorbent assay （ELISA）. Left lung tissue was used for HE staining to evaluate the pathological changes. We took the upper lobe of the right lung to extract protein; Western blot was used to detect the expression of Wnt5a protein expression; lower lobe of the right lung was used for mRNA extraction, and real-time PCR were used to observe Syndecan-1 gene expression level. We evaluated the pulmonary vascular permeability by intravenous injection of Evans blue; right middle lobe of the lung tissue was used to observe impairment and changes of vascular endothelial cell by electron microscope. Re- suits In ALl group, lung alveolar wall became thickening, large number of blood cells and inflammatory cells seeped into Alveolar space. With the severity of lung injury, the concentration of Wnt5a and Syndecan-1 in serum reached the peak at 5 h and 3 h respectively, and maintaining a higher serum concentration within 24 h. With the degree of lung injury aggravated, Wnt5a protein expression gradually increased. At the 3 h, Syndecan- 1 mRNA expression was upregulated. Formamide concentrations of Evans blue gradually increased compared with the control group. Pulmonary vascular endothelial cells swelled significantl

关 键 词：WNT5A 多配体蛋白聚糖(Syndecan-1) 脂多糖(LPS) 脓毒症 急性肺损伤(ALI) 液体管理 

分 类 号：R735.2[医药卫生—肿瘤]