骨髓间充质干细胞对幼鼠急性肺损伤模型肺组织病理学改变及肺组织白细胞介素6和8表达的影响  

Effect of bone mesenchymal stem cells on pathological changes and expression of interleukin-6 and-8 in the lung tissue in acute lung injury model of juvenile mice

作　　者：孙鹏[1,2] 李玖军[1] 刘凯[1] 张涛[1] 庄倩[1] 李倩影[1] Sun Peng;Li Jiujun;Liu Kai;Zhang Tao;Zhuang Qian(Department of PICU,Shengjing Hospital of China Medical University,Shenyang 110004,China)

机构地区：[1]中国医科大学附属盛京医院PICU,沈阳110004 [2]沈阳市儿童医院

出　　处：《中国小儿急救医学》2018年第8期624-629,共6页Chinese Pediatric Emergency Medicine

基　　金：辽宁省自然科学基金（2014021077,20092099）;辽宁省教育厅基金（2009A782）

摘　　要：目的 测定幼鼠雾化吸入脂多糖（lipopolysaccharide,LPS）所致急性肺损伤（acute lung injury,ALI）模型中肺组织表达白细胞介素6（interleukin 6,IL-6）及IL-8的变化.观察幼鼠ALI模型经骨髓间充质干细胞（bone mesenchymal stem cells,BMSCs）干预治疗后肺组织损伤程度与模型组比较有无减轻.方法 SPF级雄性SD幼鼠72只,随机分为对照组、模型组、治疗组,每组再随机分为6 h、12 h、24 h、48 h 4个亚组（每组6只）.模型组经雾化吸入LPS（5 mg/ml,2 ml,30 min）制备幼鼠ALI模型,对照组同期雾化吸入等容积生理盐水（2 ml,30 min）,治疗组在雾化吸入LPS（5 mg/ml,2 ml,30 min）后1 h,尾静脉注射BMSCs（1×106/ml）0.1 ml/kg,模型组、对照组同时间尾静脉注射等体积（0.1 ml/kg）的Dulbecco改良培养基.测定肺湿重/干重（W/D）比值,观察肺组织病理变化程度,显微镜下计数各时点支气管肺泡灌洗液（bronchoalveolar lavage fluid,BALF）中细胞数量,BCA法测定蛋白浓度,并应用Western Blot试验检测肺组织液中IL-6、IL-8蛋白含量变化.结果 模型组各时点幼鼠肺W/D比值较对照组明显增高;治疗组各时点幼鼠肺W/D比值较模型组明显下降（P〈0.05）.组织病理学观察显示：模型组幼鼠肺组织呈肺泡弥漫性充血、出血,肺组织水肿,肺泡腔和血管壁可见明显的炎性细胞浸润,肺泡壁明显增厚等明显肺损伤改变;治疗组幼鼠肺组织损伤程度较模型组明显减轻.模型组幼鼠各时点BALF中细胞数较对照组明显增高;治疗组幼鼠各时点BALF中细胞数较模型组明显下降（P〈0.05）.Western Blot结果显示：模型组各时间点幼鼠肺组织中IL-6、IL-8蛋白表达较对照组明显增高（P〈0.05）,治疗组肺组织中IL-6、IL-8蛋白表达较模型组显著降低.结论 雾化吸入LPS致幼鼠ALI模型肺组织IL-6、IL-8蛋白表达增高.BMSCs干预下,6 h、12 h、24 h、48 h幼鼠肺组织IL-6、IL-8蛋白表达较模型组降低Objective To detect the changes in expression levels of interleukin（ IL）-6 and IL-8 in the lungs of juvenile mice with acute lung injury（ ALI） induced by aerosol inhalation of lipopolysaccharide （ LPS） . To observe the degree of reparation of lung damage after intervening with exogenous bone mesenchy-mal stem cells （ BMSCs） . Methods Seventy-two SPF grade male SD juvenile mice were randomly divided into 3 groups：control group,model group,and treatment group. Each group was randomly divided into 4 sub-groups at 6 h,12 h,24 h,and 48 h after the building of model（6 mice per group）. The model group were in-haled by LPS （5 mg/ml,2 ml,30 min） to prepare the ALI model. The control group were inhaled by the same volume of physiological saline （2ml,30min） during the same period. As for the treatment group, BMSCs was injected into the tail vein one hour after inhalation of LPS （1 × 106/ml,0. 1 ml/kg）. Mean-while,the model group and the control group were injected intravenously with equal volume （0. 1 ml/kg） of Dulbecco＇s modified eagle medium. The ratioes of wet weight and dry weight of the lungs（ W/D） were detec-ted. The pathological changes in lungs were observed by optical microscope. The numbers of cells in the bron-choalveolar lavage fluid （ BALF） at each time point were counted under a microscope and the protein con-centrations were measured by bicinchoninic acid method. The changes of IL-6 and IL-8 protein content in lung tissue fluid were measured by applying Western Blot method. Results Compared with the control group,the ratioes of W/D in the lung of mice at each time piont of the model group were significantly high-er;the W/D ratioes of the lungs in the treatment group were significantly lower than those of model group （P〈0. 05）. In accordance with histopathological observations,the lung tissue of the juvenile mice in the model group showed alveolar diffuse congestion, hemorrhage, pulmonary edema, obvious inflammatory cell infiltration in the alveolar cavity

关 键 词：骨髓间充质干细胞 脂多糖 急性肺损伤 白细胞介素6 白细胞介素8 

分 类 号：R725.6[医药卫生—儿科] R-332[医药卫生—临床医学]