摘要
目的本研究旨在探讨靶向DEK适配体DTA-64在脂多糖(LPS)诱导的小鼠急性肺损伤(ALI)中的保护作用。方法将雌性Balb/c小鼠,随机分为4组,PBS组、LPS诱导的ALI模型组、LPS+对照适配体组和LPS+DTA 64干预组,每组8只。通过HE染色观察肺组织病理改变;ELISA和流式细胞术检测血清和支气管肺泡灌洗液(BALF)的TNF-α,IL-1β表达;采用Western blot法检测GSK-3β/β-catenin和HIF1-α/MMP-9信号通路表达,探讨DTA-64在急性肺损伤中的作用机制。结果DEK蛋白在ALI模型鼠肺组织中表达增强,在DTA-64治疗组中表达降低;DTA-64能够降低肺组织中炎性细胞浸润,下调肺组织损伤病理评分和肺湿/干质量比、炎性细胞数量以及MPO在肺组织中的表达和活性;DTA-64还能降低血清、BALF和纵膈淋巴结中TNF-α和IL-1β水平;DTA-64下调磷酸化GSK-3β表达、β-catenin、CyclinD1、C-Jun、C-Myc、HIF1-α、VEGF、MMP-9和MMP-2表达。结论DTA-64可能通过GSK-3β/β-catenin和HIF1-α/MMP-9信号通路抑制LPS诱导的ALI小鼠肺部炎症细胞聚集和病理损伤。这有望成为临床治疗ALI的新的靶点。
The aim of this study was to investigate the inhibitory effect of DEK-targeting aptamer DTA-64 on lipopolysaccharide(LPS)-induced acute lung injury(ALI)of mice.Thirty-two female Balb/c mice were randomly divided into 4 groups:PBS group,ALI model group,LPS+DTA-64 treated group and LPS+control aptamer group,with 8 mice in each group.Inflammatory cell infiltration in lung tissues were detected by HE staining;pathological injury of lung tissues was evaluated;TNF-αand IL-1βexpression in the serum,BALF and lymph nodes was detected by ELISA or by flow cytometry;the expression of GSK-3β/β-catenin and HIF1-α/MMP-9 signaling pathway-related proteins were detected by Western blotting to investigate the mechanism of DTA-64 in acute lung injury.Data showed that DEK protein was increased in the lung tissue of acute lung injury model mice,and reduce inflammatory cell infiltration in the lung tissue,down-regulate the pathological score of lung tissueinjury and lung Wet/Dry weight ratio,inflammatory cellnumbers and the MPO activity in the lung tissue;DTA-64 could also reduce TNF-αand IL-1βlevels in serum,BALF and mediastinal lymph nodes;furthermore,DTA-64 down-regulated phosphorylated GSK-3βexpression,β-catenin,CyclinD1,C-Jun,C-Myc,HIF1-α,VEGF,MMP-9 and MMP-2 expression.In conclusion,DTA-64 may inhibit inflammatory cell accumulation andpathological damage in the lungs of mice with LPS-induced acute lung injury through the GSK-3β/β-catenin andHIF1-α/MMP-9signalingpathways,which would beexpected to beanew target for clinical treatment of acute lung injury.
作者
宋艺兰
白巧云
韩雪
马小斐
王丹丹
朴红梅
李良昌
延光海
SONG Yilan;BAI Qiaoyun;HAN Xue;MA Xiaofei;WANG Dandan;PIAO Hongmei;LI Liangchang;YAN Guanghai(Jilin Key Laboratory for Immune and Targeting Research on Common Allergic Diseases,Yanbian University,Yanji 133002,China;Department of Anatomy,Histology and Embryology,Yanbian University Medical College,Yanji 133002,China;Department of Respiratory Medicine,Yanbian University Hospital,Yanji 133000,China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2022年第10期862-868,875,共8页
Immunological Journal
基金
国家自然科学基金项目(82160004,81970018,81860729)
中央引导地方科技发展资金吉林省基础研究专项(202002020JC)。
