短链脂肪酸诱导巨噬细胞极化减轻脓毒症急性肺损伤  

Macrophage polarization induced by short-chain fatty acid attenuates acute lung injury in sepsis

作　　者：马金兰 杨舸 马玉杰[1] 刘勤富 杨晓军[1] Ma Jinlan;Yang Ge;Ma Yujie;Liu Qinfu;Yang Xiaojun(Department of Critical Care Medicine,General Hospital of Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China)

机构地区：[1]宁夏医科大学总医院重症医学科,银川750004

出　　处：《中华危重病急救医学》2023年第9期933-938,共6页Chinese Critical Care Medicine

基　　金：宁夏回族自治区自然科学基金(2023AAC03559)。

摘　　要：目的探讨短链脂肪酸(SCFA)通过巨噬细胞极化对脓毒症急性肺损伤(ALI)的影响.方法①临床试验:纳入宁夏医科大学总医院重症监护病房(ICU)2022年1月至12月收治的30例脓毒症患者及同期10例非脓毒症患者,收集患者入院第1天粪便样本,利用靶向代谢组学检测两组患者SCFA丁酸水平.②动物实验:选取雄性C57BL/6J小鼠,按随机数字表法分为假手术组(Sham组)、盲肠结扎穿孔术致脓毒症组(CLP组)和SCFA干预组(SCFA组,CLP后1h灌胃丁酸钠25 mg/kg),每组6只.制模后24 h,利用小鼠脓毒症评分(MSS)评估小鼠状态,计算肺湿/干质量(W/D)比值评估肺水肿程度,苏木素-伊红(HE)染色观察肺组织病理学改变并进行肺损伤评分,用酶联免疫吸附试验(ELISA)检测血清炎症因子肿瘤坏死因子-α(TNF-α)、白细胞介素(IL-1β、IL-6、IL-10)、核转录因子-κB(NF-κB)、转化生长因子-β(TGF-β)水平,用实时荧光定量聚合酶链反应(RT-qPCR)测定肺组织炎症因子TNF-α、IL-1β、IL-6以及抗氧化因子核因子E2相关因子2(Nrf2)的mRNA表达,免疫组化法检测肺组织巨噬细胞极化标志物精氨酸-1(ARG-1)、CD206、诱导型一氧化氮合酶(iNOS)和IL-1β的表达.结果①与非脓毒症患者相比,脓毒症患者SCFA丁酸水平显著下降(μg/g:34.56±6.61比1150.67±381.90,P<0.01).②与Sham组相比,CLP组小鼠MSS评分、肺W/D比值、肺损伤评分,血清炎症因子TNF-α、TGF-β、NF-κB、IL-10、IL-6、IL-1β水平,肺组织炎症因子和抗氧化因子Nrf2的mRNA表达,以及M1型巨噬细胞极化标志物ARG-1、CD206和M2型巨噬细胞极化标志物iNOS、IL-1β表达明显升高.与CLP组相比,SCFA组小鼠MSS评分、肺W/D比值、肺损伤评分、血清炎症因子TNF-α、TGF-β、NF-κB、IL-10、IL-6、IL-1β水平、肺组织炎症因子和抗氧化因子Nrf2的mRNA表达,以及M1型巨噬细胞极化标志物ARG-1、CD206表达均明显降低[MSS评分(分):14.50±3.16比20.00±1.55,肺W/D比值:4.60±0Objective To explore the effect of short-chain fatty acid(SCFA)on acute lung injury(ALI)in sepsis via macrophage polarization.Methods①Clinical trial:30 sepsis patients admitted to the intensive care unit(ICU)of General Hospital of Ningxia Medical University from January to December in 2022 and 10 non-sepsis patients in the same period were enrolled,stool samples were collected on the first day of admission,and SCFA butyric acid level in the two groups were studied by targeted metabolomics.②Animal experiment:male C57BL/6J mice were selected and randomly divided into sham operation group(Sham group),sepsis caused by cecal ligation and perforation(CLP group)and SCFA intervention group(SCFA group,sodium butyrate 25 mg/kg was given by gavage 1 hour after CLP),with 6 animals in each group.Twenty-four hours after molding,the state of mice was evaluated by mouse sepsis score(MSS),the degree of pulmonary edema was evaluated by calculating the wet/dry ratio(W/D),and the pathological changes of lung tissue were observed by hematoxylin-eosin(HE)staining,and lung injury score was performed.The serum levels of tumor necrosis factor-α(TNF-α),interleukins(IL-1β,IL-6,IL-10),nuclear factor-κB(NF-κB),and transforming growth factor-β(TGF-β)were detected by enzyme-linked immunosorbent assay(ELISA).Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to measure the mRNA expressions of inflammatory factors TNF-α,IL-1β,IL-6 and antioxidant factor nuclear factor E2-related factor 2(Nrf2)in lung tissue.The expressions of macrophage polarization markers arginin-1(ARG-1),CD206,inducible nitric oxide synthase(iNOS)and IL-1βin lung tissue were detected by immunohistochemistry.Results①Compared with non-sepsis patients,SCFA-butyric acid level was significantly reduced in patients with sepsis(μg/g:34.56±6.61 vs.1150.67±381.90,P<0.01).②Compared with the Sham group,MSS,lung W/D ratio,lung injury score,the levels of serum inflammatory factors TNF-α,TGF-β,NF-κB,IL-10,IL-6,IL-1β,the mRNA expressions

关 键 词：脓毒症 急性肺损伤 短链脂肪酸 巨噬细胞极化 

分 类 号：R459.7[医药卫生—急诊医学]