细胞外信号调节激酶信号通路在失血性休克致肺损伤中趋化因子分泌和中性粒细胞募集中的作用及其机制  

Role and mechanism of extracellular signal-regulated kinase signaling pathway in chemokine secretion and neutrophil recruitment in hemorrhagic shock-induced lung injury

作　　者：李新新[1] 肖鹏[2] 叶菁菁 李建强 周伟[2] 尚文韬 付学峥 周培松 王志伟[2,3] Li Xinxin;Xiao Peng;Ye Jingjing;Li Jianqiang;Zhou Wei;Shang Wentao;Fu Xuezheng;Zhou Peisong;Wang Zhiwei(Department of Pain Treatment,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Department of Orthopedics,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;National Trauma Medicine Center,Peking University People’s Hospital,Beijing 100044,China)

机构地区：[1]郑州大学第一附属医院疼痛科,郑州450052 [2]郑州大学第一附属医院骨科,郑州450052 [3]北京大学人民医院国家创伤医学中心,北京100044

出　　处：《中华实验外科杂志》2024年第5期999-1002,共4页Chinese Journal of Experimental Surgery

基　　金：河南省医学科技攻关计划联合共建项目(LHGJ20220356);"北京大学人民医院研究与发展基金"学术新星项目(RS2023-03)

摘　　要：目的:探讨失血性休克(HS)致急性肺损伤(ALI)中细胞外信号调节激酶(ERK)通路的改变及其抑制剂的保护作用。方法:SD大鼠分为假手术(Sham)组、HS 4 h组、HS 8 h组、干预组和溶剂对照组,每组6只。HS组和干预组采用股静脉插管定量失血建立模型,Sham组只进行手术操作。干预组和溶剂对照组在失血后20 min腹腔注射ERK蛋白磷酸化抑制剂司美替尼(Selumetinib)或等体积溶剂。细胞实验分为正常培养组、缺氧缺糖组(OGD)、Selumetinib干预组和溶剂对照组。使用肺组织苏木精-伊红(HE)染色、免疫组织化学染色、病理评估和炎性细胞计数评估肺组织损伤情况,采用酶联免疫吸附试验(ELISA)分析趋化因子含量,采用蛋白质免疫印迹分析ERK蛋白磷酸化水平。两组之间统计比较使用非配对t检验或Welch’s t检验,多组间比较使用单因素方差分析(One-Way ANOVA)或Kruskal-Wallis检验,并采用Bonferroni post-hoc检验进行组间的两两比较。结果:HS 8 h组大鼠中性粒细胞(105.58±14.73)显著多于Sham组(25.12±5.48),差异有统计学意义(t=12.536,P<0.05);HS 8 h组肺损伤评分[(0.682±0.089)分]显著高于Sham组[(0.175±0.083)分],差异有统计学意义(t=10.199,P<0.05)。HS 4 h和HS 8 h组肺组织的CXCL1浓度和CXCL2浓度显著高于Sham组,差异有统计学意义(CXCL1:F=89.38,P<0.01;CXCL2:χ^(2)=15.158,P<0.01)。HS 4 h和HS 8 h组血清的CXCL1和CXCL2浓度显著多于假手术组,差异有统计学意义(CXCL1:χ^(2)=14.000,P<0.01;CXCL2:χ^(2)=15.158,P<0.01)。OGD培养肺上皮细胞(A549细胞)2、4、6 h后,IL-8、CXCL1和CXCL2含量显著增多(IL-8:χ^(2)=21.60,P<0.01;CXCL1:F=341.043,P<0.01;CXCL2:χ^(2)=17.78,P<0.01),同时,ERK蛋白磷酸化水平逐渐显著增强(t=6.867,P<0.05),Selumetinib显著抑制A549细胞趋化因子IL-8(t=10.497,P<0.01)、CXCL1(t=10.631,P<0.01)和CXCL2(t=5.504,P<0.01)的分泌。动物实验中,Selumetinib抑制CXCL1/2分泌(P<0.01)和中性粒细胞浸润(t=9.731,P<0.01),减Objective To investigate the change of extracellular signal-regulated kinase(ERK)pathway and the protective effect of hemorrhagic shock(HS)inhibitors in acute lung injury(ALI).Methods SD rats were divided into the Sham group,HS 4 h group,HS 8 h group,intervention group,and solvent control group with 6 rats in each group.The HS group and the intervention group were set up with quantitative blood loss by femoral vein intubation,while the Sham group was only operated.The intervention group and solvent control group were intraperitoneally injected with ERK protein phosphorylation inhibitor Selumetinib or equal-volume solvent 20 min after blood loss.Cell experiments were divided into normal culture group,oxygen-glucose deprivation(OGD)group,Selumetinib intervention group,and solvent control group.Lung tissue injury was evaluated by hematoxylin-eosin(HE)staining,immunohistochemical staining,pathological examination,and inflammatory cell count.Chemokine content was analyzed by enzyme-linked immunosorbent assay(ELISA),and ERK protein phosphorylation level was analyzed by Western blotting.The unpaired t test or Welch’s t test was used for statistical comparison between the two groups,One-Way ANOVA or Kruskal-Wallis test was used for multi-group comparison,and the Bonferroni post-hoc test was used for pair-to-group comparison.Results Neutrophils in the HS 8 h group(105.58±14.73)were significantly more than those in the Sham group(25.12±5.48,t=12.536,P<0.05).The lung injury score in the HS 8 h group[(0.682±0.089)points]was significantly higher than that of the Sham group[(0.175±0.083)points,t=10.199,P<0.05].The concentration of CXCL1 and CXCL2 in lung tissue of HS 4 h and HS 8 h groups was significantly higher than that of Sham group(CXCL1:F=89.38,P<0.01;CXCL2:χ^(2)=15.158,P<0.01).Serum CXCL1 and CXCL2 concentrations in HS 4 h and HS 8 h groups were significantly higher than those in sham operation group(CXCL1:χ^(2)=14.000,P<0.01;CXCL2:χ^(2)=15.158,P<0.01).The contents of IL-8,CXCL1,and CXCL2 increased significantl

关 键 词：失血性休克 急性肺损伤 细胞外信号调节激酶信号通路 趋化因子 中性粒细胞 

分 类 号：R563[医药卫生—呼吸系统]