人羊膜上皮细胞气管内滴注对脂多糖诱导小鼠急性肺损伤的保护作用  

Protective effect of human amnion epithelial cells through endotracheal instillation against lipopolysaccharide-induced acute lung injury in mice

作　　者：夏彦民 王文辰[1] 张鸿[2] 陈家宽[1] 王明[1] 葛忠虎[1] 汪健[1] 姜涛[1] 

机构地区：[1]第四军医大学附属唐都医院胸腔外科,陕西西安710038 [2]陕西省人民医院血管外科,陕西西安710068

出　　处：《细胞与分子免疫学杂志》2017年第1期7-11,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：陕西省自然科学基础研究计划项目(2014JM2-8144)

摘　　要：目的探讨人羊膜上皮细胞(hAEC)对脂多糖(LPS)诱导的急性肺损伤小鼠的治疗作用及其相关机制。方法雄性C57BL/6小鼠随机分成正常对照组、hAEC对照组、LPS诱导的急性肺损伤组和hAEC治疗组,每组10只。后2组小鼠经口咽插管注入LPS(5 mg/kg),1 h后,再分别注入PBS(50μL)或1×107个/m L hAEC(50μL);正常对照组和hAEC对照组则经口咽插管注入等量PBS或hAEC。LPS处理24 h后处死小鼠,收集血液及肺脏组织,提取肺组织细胞质/细胞核蛋白,另取肺组织进行石蜡包埋、切片;称量并计算肺组织湿/干质量(W/D)比;ELISA检测血清中白细胞介素10(IL-10)、IL-6、IL-1β及肿瘤坏死因子α(TNF-α)含量;HE染色观察肺组织病变,Western blot法检测肺组织核转录因子κBp65(NF-κBp65)蛋白的表达。结果与正常对照组相比,LPS处理后小鼠肺组织W/D比增大,血清中IL-6、IL-1β及TNF-α含量增加,而IL-10含量减少;肺组织微血管破坏,发生肺水肿,肺泡间隔增厚;肺组织中NF-κBp65蛋白活性增加。hAEC处理使肺组织病变减轻,W/D比减小,血清中IL-6、IL-1β及TNF-α含量减少,肺组织NF-κBp65蛋白活性降低。结论 hAEC可降低肺组织中NF-κBp65的活性,显著减轻LPS诱导的急性肺损伤。Objective To investigate the therapeutic effect of human amnion epithelial cels（ hAECs） through endotracheal instillation on acute lung injury（ ALI） induced by lipopolysaccharide（ LPS） in mice and its underlying mechanism. Methods Male C57BL/6 mice were randomly divided into normal control group,hAECs control group,LPS-induced ALI group and hAECs treatment group（ n = 10 each group）. In the latter two groups,mice were subjected to LPS（ 5 mg/kg） by oral insertion,and 1 hour later,PBS（ 50 μL） and hAECs（ 1 × 107/m L,50 μL） were administered by oral insertion in the two groups,respectively. The normal control group and hAECs control group were injected with the same amount of PBS and hAECs by oral insertion,respectively. Twenty-four hours after LPS administration,the mice were sacrificed to collect blood and lung tissues. Cytoplasmic and nuclear proteins were extracted from the lung tissues,and other lung tissues were embedded in paraffin and cut into sections. The wet mass and dry mass of lung tissues were obtained to calculate the wet/dry mass ratios. Concentrations of interleukin（ IL）-10,IL-6,IL-1β and tumor necrosis factor（ TNF）-α in sera were measured by ELISA. Pulmonary histological changes were evaluated by HE staining. The nuclear transcription factor-kappa Bp65（ NF-κBp65）activation in lung tissues was detected by Western blotting. Results Compared with the control group,the lung wet/dry mass ratios and the serum concentrations of TNF-α,IL-1β and IL-6 in the LPS-induced model group increased,while the level of IL-10 decreased. Under a light microscope,the lung tissues from LPS-induced model group showed serious pulmonary edema,haemorrhagia in stroma,alveolar wall thickness,alveolus collapse and obvious inflammatory cell infiltration. LPS administration also increased the activation of NF-κBp65 in lung tissues. hAECs administration significantly decreased LPS-induced lung histopathological changes,lung wet/dry mass ratios and the serum levels of IL-6,IL-1β and T

关 键 词：急性肺损伤 脂多糖 人羊膜上皮细胞(hAEC) 核转录因子κBp65(NF-κBp65) 

分 类 号：R563[医药卫生—呼吸系统]