甘草酸二铵脂质配位体抑制内毒素引起的急性肺损伤中炎性细胞浸润及细胞间连接蛋白的降解  

Diammonium glycyrrhizinate lipid ligand inhibits endotoxin induced inflammatory cell infiltration and cell junctional protein degradation in acute lung injury

作　　者：刘梅梅[1] 王齐[1] 杨珺[1] 黄焱平 周瑾[1] Liu Meimei;Wang Qi;Yang Jun;Huang Yanping;Zhou Jin(Department of Histology and Embryology, Anhui Medical College, Hefei 230601)

机构地区：[1]安徽医学高等专科学校人体解剖学与组织胚胎学教研室,合肥230601

出　　处：《中国组织化学与细胞化学杂志》2017年第6期546-553,共8页Chinese Journal of Histochemistry and Cytochemistry

基　　金：安徽省自然科学研究重点项目(KJ2016A374)

摘　　要：目的探讨甘草有效成分甘草酸二铵脂质配位体(diammonium glycyrrhizinate lipid ligand,DGLL)对内毒素(lipopolysaccharide,LPS)诱发的肺损伤和肺水肿的改善作用及其机制。方法本研究利用腹腔给入LPS(10mg/kg)建立大鼠急性肺损伤(acute lung injury,ALI)模型,在LPS诱导前1小时灌胃给入DGLL(30、60、120mg/k g)。在LPS诱导6小时后,用HE组织学染色方法评价肺损伤,用肺湿干重比、肺泡灌洗液蛋白含量检测和肺组织伊文思蓝渗出法综合评价肺水肿,用ELISA方法检测肺组织中炎性因子和粘附分子含量,用免疫组织化学染色方法检测肺组织白细胞浸润标志物髓过氧化物酶(myeloperoxidase,MPO)的表达,用Western blot方法检测炎症相关蛋白以及肺微血管通透性相关蛋白水平。结果 DGLL前给药可以显著抑制LPS诱导的肺组织损伤,降低LPS诱导的MPO免疫反应性增强,降低肺组织中炎性因子TNF-α、IL-6和粘附分子ICAM-1的表达;同时,DGLL可以抑制LPS诱导的肺水肿、降低肺泡灌洗液中的蛋白含量以及肺组织伊文思蓝的渗出;抑制血管内皮钙黏蛋白(vascular endothelial cadherin,VE-cadherin)和ZO-1、Occludin、JAM-1等紧密连接蛋白的降解。结论 DGLL对LPS诱导的大鼠ALI具有明显的抑制作用,其作用机制与抑制炎性细胞的浸润以及细胞间连接蛋白的降解相关。该结果为D GLL在临床治疗ALI提供了新的理论依据。Objective To explore the mechanism of diammonium glycyrrhizinate lipid ligand(DGLL)in improving lipopolysaccharide(LPS)induced acute lung injury and pulmonary edema.Methods In the study,rat acute lung injury(ALI)model was induced by intraperitonreal injection of LPS(10mg/kg).DGLL(30,60,120mg/kg)was administrated by gastric gavage one hour prior.Six hours later,pathological changes of lung injury were evaluated by HE staining,while the degree of pulmonary edema was assessed by tissue wet-to-dry weight ratio,protein content in bronchoalveolar lavage(BAL)fluid and the accumulation of Evans blue dye in alveolar space.Inflammatory cytokines and adhesion molecules in injured lung tissues were quantified by ELISA or Western blot.In addition,immunohistochemistry was used to identify leukocyte infiltration marker myeloperoxidase(MPO).Result DGLL significantly reduced the number of MPO positive cells and the expressions of TNF-α,IL-6I and CAM-1in injured lung tissues.Meanwhile,DGLL appeared to relieve pulmonary edema indicated by decreased protein content in BAL fluid and Evans blue in alveolar space.DGLL also inhibited the degradation of vascular endothelial cadherin(VE-Cadherin)and tight junction proteins ZO-1,Occludin,and JAM-1.Conclusion DGLL demonstrated a significant inhibitory effect on LPS-induced rat ALI.The mechanism was likely related to its inhibition of inflammatory cell infiltration and cell junctional protein degradation.The results provided a new theoretical basis for the clinical treatment of ALI by DGLL.

关 键 词：甘草酸二铵脂质配位体 炎性浸润 血管内皮钙黏蛋白 紧密连接蛋白 

分 类 号：R563.9[医药卫生—呼吸系统]