RANKL在脓毒症相关急性肺损伤炎症反应中的作用及机制  

Mechanism of action of RANKL in inflammatory response to sepsis-associated acute lung injury

作　　者：牛新荣 陈蔚霖[1] 李辉 NIU Xinrong;CHEN Weilin;LI Hui(Graduate School of Xinjiang Medical University,Urumqi 830017,China;Second Department of Critical Care Medicine,People’s Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830001,China)

机构地区：[1]新疆医科大学研究生学院,新疆乌鲁木齐830017 [2]新疆维吾尔自治区人民医院重症医学二科,新疆乌鲁木齐830001

出　　处：《安徽医学》2025年第2期135-140,共6页Anhui Medical Journal

基　　金：新疆维吾尔自治区自然科学基金(编号:2022D01C604)。

摘　　要：目的探讨核因子-κB受体活化因子配体(RANKL)的预处理对脓毒症相关急性肺损伤(SA-ALI)中的影响,并探究其通过炎症反应发挥作用的可能机制。方法将24只雄性C57BL/6小鼠随机分为假手术(Sham)组、盲肠结扎穿孔(CLP)组,RANKL组与抗RANKL组,每组6只。每组小鼠给予预处理,RANKL组、抗RANKL组经腹腔注射重组RANKL、抗RANKL抗体,Sham组、CLP组给予等量的生理盐水。1 h后Sham组打开腹部翻转盲肠后闭合腹部,其余各组使用CLP法建立SA-ALI模型。造模24 h后,处死小鼠并取材。HE染色法检测肺组织病理变化;酶联免疫吸附(ELISA)试验检测血清中白细胞介素-6(IL-6)、肿瘤坏死因子α(TNF-α)和白细胞介素-1β(IL-1β)含量;实时荧光定量逆转录聚合酶链式反应(RT-qPCR)及蛋白免疫印迹法(WB)检测肺组织RANKL、核因子-κB受体活化因子受体(RANK)、Toll样受体4(TLR4)mRNA及蛋白表达水平;结果在CLP诱导的SA-ALI小鼠模型中,肺组织内炎性细胞浸润明显,血清中IL-6、TNF-α和IL-1β水平较Sham组增高(P<0.05)。RANKL预处理后肺组织病理损伤减轻,血清中IL-6、TNF-α和IL-1β的含量和肺组织中的RANK、TLR4的蛋白及mRNA的表达较CLP组降低,RANKL蛋白及mRNA的表达升高(P<0.05)。而抗RANKL处理后,小鼠的肺组织病理损伤加重,血清中IL-6、TNF-α和IL-1β的含量和肺组织中的RANK、TLR4的蛋白及mRNA的表达较CLP组升高,RANKL蛋白及mRNA的表达降低,差异均具有统计学意义(P<0.05)。结论RANKL的预处理可通过调节炎症反应缓解SA-ALI小鼠的肺损伤,其机制可能与RANKL/RANK/TLR4信号通路有关。Objective To investigate the effect of pretreatment with receptor activator of nuclear factor kappa B ligand(RANKL)on sepsis associated acute lung injury(SA-ALI)and explore its possible mechanism of action through inflammatory response.Methods Twenty-four male C57BL/6 mice were randomly divided into sham operation group,CLP group,RANKL group,and anti RANKL group.Each group consists of six animals.Each group of mice was given pre-treatment.The RANKL group and anti RANKL group were intraperitoneally injected with recombinant RANKL and anti RANKL antibodies,while the sham surgery group and CLP group were given equal amount of physiological saline.One hour later,the sham surgery group opened the abdomen,flipped the cecum,and closed the abdomen.The other groups used cecal li-gation and puncture(CLP)to establish the SA-ALI model.After 24 hours of modeling,the mice were euthanized and samples were taken.HE staining method was used to detect pathological changes in lung tissue;Enzyme linked immunosorbent assay was used to detect the levels of IL-6,TNF-α,and IL-1βin serum;Real time fluorescence quantitative PCR and protein immunoblotting were used to detect the mRNA and pro-tein expression levels of RANKL,RANK,and Toll like receptor 4(TLR4)in lung tissue.Results In the CLP induced SA-ALI mouse model,there was significant infiltration of inflammatory cells in the lung tissue,and the levels of IL-6,TNF-α,and IL-1βin the serum increased compared to the sham operation group.After RANKL pretreatment,the pathological damage to lung tissue was reduced,and the levels of IL-6,TNF-α,and IL-1βin serum,as well as the protein and mRNA expression of RANK and TLR4 in lung tissue decreased compared to the CLP group.The expression of RANKL protein and mRNA increased.After treatment with anti RANKL,the pathological damage to the lung tissue of mice worsened,and the levels of IL-6,TNF-α,and IL-1βin serum,as well as the protein and mRNA expression of RANK and TLR4 in lung tissue increased compared to the CLP group.The expression of R

关 键 词：脓毒症 急性肺损伤 核因子-κB受体活化因子受体 核因子-ΚB受体活化因子配体 TOLL样受体4 

分 类 号：R459.7[医药卫生—急诊医学]