TNFR-Fc对急性肺损伤炎症失衡中MAPK通路的影响  

Effect of TNFR-Fc on MAPK pathway in the inflammatory imbalance of acute lung injury

作　　者：袁伟锋 李理 黄文杰 郑燕列 YUAN Wei-feng;LI Li;HUANG Wen-jie;ZHENG Yan-lie(Department of Respiratory Medicine,Guangzhou Eighth People′s Hospital of Guangzhou Medical University,Guangzhou,Guangdong 510060,China;Department of Infectious Disease,General Hospital of Southern Theater Command of PLA,Guangzhou,Guangdong 510010,China;Department of Pulmonary and Critical Care Medicine,General Hospital of Southern Theater Command of PLA,Guangzhou,Guangdong 510010,China)

机构地区：[1]广州医科大学附属市八医院呼吸内科,广东广州510060 [2]中国人民解放军南部战区总医院感染科,广东广州510010 [3]中国人民解放军南部战区总医院呼吸与危重症医学科,广东广州510010

出　　处：《临床肺科杂志》2022年第7期1051-1054,1060,共5页Journal of Clinical Pulmonary Medicine

基　　金：国家自然科学基金(No.81200002,No.81370173);广州市科技计划项目(No.201707010020);广东省医学科学技术研究基金项目(No.B2021027)。

摘　　要：目的评价在脂多糖(LPS)导致的炎症反应失衡中,肿瘤坏死因子受体Fc段融合蛋白(TNFR-Fc)对丝裂原活化蛋白激酶(MAPKs)磷酸化水平的影响。方法24只BALB/c小鼠随机平均分为LPS组与TNFR-Fc+LPS组。气管内滴入LPS复制急性肺损伤(ALI)的炎症失衡小鼠模型,腹腔内注射TNFR-Fc中和肿瘤坏死因子(TNF-α),2小时后收集标本,ELISA法检测血清/支气管肺泡灌洗液(BALF)中白细胞介素-1β(IL-1β)、IL-6、IL-10、干扰素-γ(IFN-γ)浓度,RT-PCR法检测肺组织TNF-α与核因子-κB(NF-κB)基因转录强度,Western Blot检测肺组织MAPKs(包括Erk1/2、p38与JNK)磷酸化水平。结果两组小鼠在气管内滴入LPS后血清与BALF中IL-6浓度显著增高,TNF-α转录水平增高,MAPKs磷酸化水平增高;TNFR-Fc+LPS组小鼠血清与BALF中IL-6浓度较LPS组显著降低,TNF-α转录水平下降,MAPK磷酸化水平下降。结论TNFR-Fc中和TNF-α能下调炎症反应信号通路中MAPKs磷酸化水平,下调炎症反应强度,恢复ALI小鼠的炎症反应失衡。Objective To evaluate the effect of tumor necrosis factor receptor Fc region fusion protein(TNFR-Fc)on the phosphorylation level of mitogen-activated protein kinases(MAPKs)in the imbalance of inflammatory response induced by lipopolysaccharide(LPS).Methods A total of 24 BALB/c mice were randomly divided into the LPS group and TNFR-Fc+LPS group.Intratracheal instillation of LPS was used to replicate the inflammatory imbalance mouse model of acute lung injury(ALI),and TNFR-Fc was administered via intraperitoneal injection to neutralize tumor necrosis factor(TNF-α).The samples were collected 2 hours later,and the concentrations of interleukin-1β(IL-1β),IL-6,IL-10,and interferon-γ(IFN-γ)in serum/bronchioalveolar lavage fluid(BALF)were detected by ELISA.RT-PCR was used to detect the transcriptional intensity of TNF-αand nuclear factor-κB(NF-κB)genes in lung tissue,and Western Blot was used to detect the phosphorylation levels of MAPKs(including Erk1/2,p38,and JNK)in lung tissue.Results The concentration of IL-6 in serum and BALF,TNF-αtranscription level,and MAPKs phosphorylation level of mice in both groups were significantly increased after LPS administration.Compared with the LPS group,IL-6 concentration in serum and BALF of mice in the TNFR-FC+LPS group was significantly decreased,TNF-αtranscription level was decreased,and MAPK phosphorylation level was decreased.Conclusion Neutralization of TNF-αby TNFR-Fc can down-regulate the phosphorylation level of MAPKs in the inflammatory response signaling pathway,down-regulate the inflammatory response intensity,and restore the inflammatory response imbalance in ALI mice.

关 键 词：炎症反应 肿瘤坏死因子受体Fc段融合蛋白 裂原活化蛋白激酶 急性肺损伤 

分 类 号：R563[医药卫生—呼吸系统]