FIRRE在脂多糖诱导急性肺损伤中的作用及机制研究  

Role and mechanism of FIRRE in LPS-induced acute lung injury

作　　者：赵晓霞 李晓梅[1] 桑义 许从磊 李昭伦 孙文静[1] 刘晓明 Zhao Xiaoxia;Li Xiaomei;Sang Yi;Xu Conglei;Li Zhaolun;Sun Wenjing;Liu Xiaoming(Department of Pediatrics,Binzhou Medical University Hospital,Binzhou 256600,China)

机构地区：[1]滨州医学院附属医院儿科,滨州256600

出　　处：《国际医药卫生导报》2024年第20期3351-3356,共6页International Medicine and Health Guidance News

基　　金：山东省省级临床重点专科学科建设(SLCZDK-1405儿童重症科);2021年度滨州医学院“临床+X”项目(BY2021LCX29)。

摘　　要：目的探讨FIRRE在脂多糖(LPS)诱导急性肺损伤(ALI)中的作用及机制。方法研究时间为2020年6月至2022年5月,地点为滨州医学院附属医院。第一部分:FIRRE在LPS诱导A549细胞损伤中的作用研究。A549细胞分为4组,对照组采用生理盐水,低分子肝素钙组采用5 IU/ml低分子肝素钙,LPS组采用10 mg/L LPS,LPS+低分子肝素钙组采用10 mg/L LPS+5 IU/ml低分子肝素钙;处理24 h。采用CCK-8检测各组细胞活性;采用酶联免疫吸附试验(ELISA)检测白细胞介素(IL)-6、IL-10和肿瘤坏死因子(TNF-α)的表达;采用实时定量PCR检测FIRRE基因的相对表达量。第二部分:FIRRE在LPS诱导C57BL/6J小鼠ALI中的作用研究。将24只健康清洁SPF级雄性C57BL/6J小鼠(体质量25~30 g,6~8周)随机分为3组,对照组腹腔注射等体积生理盐水,ALI组注射5 mg/kg LPS诱发ALI,低分子肝素钙组腹腔注射5 mg/kg LPS诱发ALI+低分子肝素钙5 AXa IU/kg。分别于6 h、12 h评估各组小鼠毛色、反应能力、对抗外界阻力等变化;肺组织切片HE染色观察肺组织损伤程度,并进行肺损伤评分;腹主静脉取血,采用ELISA检测血清IL-6、IL-10的表达情况;采用ELISA检测肺泡灌洗液IL-6、IL-10含量;采用RT-PCR检测FIRRE基因的相对表达量。采用单因素方差分析、LSD-t检验、Dunnett-t检验。结果第一部分:LPS组A549细胞活性低于对照组,LPS+低分子肝素钙组A549细胞活力高于LPS组,多组间比较差异有统计学意义(F=44.25,P<0.05)。LPS组、LPS+低分子肝素钙组IL-6、IL-10、TNF-α水平均高于对照组、低分子肝素钙组,LPS+低分子肝素钙组IL-6、IL-10、TNF-α水平均低于LPS组(均P<0.05)。LPS组、LPS+低分子肝素钙组A549细胞FIRRE表达水平均高于对照组,LPS+低分子肝素钙组低于LPS组,多组间比较差异有统计学意义(F=242.23,P<0.001)。第二部分:ALI组及低分子肝素钙组小鼠反应能力、对抗外界阻力能力均低于对照组。LPS注射后6 h、12 h,ALI组HE染色肺Objective To explore the role and mechanism of FIRRE in lipopolysaccharide(LPS)induced acute lung injury(ALI).Methods The study was conducted at Binzhou Medical University Hospital from June 2020 to May 2022.Part I:The role of FIRRE in LPS-induced A549 cell injury.A549 cells were divided into a control group,a low molecular weight heparin calcium(LMWH)group,a LPS group,and a LPS+LMWH group,which were treat by normal saline,5 IU/ml LMWH,10 mg/L LPS,and 10 mg/L LPS+5 IU/ml LMWH for 24 h.The cell activity of each group was detected by CCK-8,the expressions of interleukin(IL)-6,IL-10,and tumor necrosis factor(TNF)-αwere detected by enzyme-linked immunosorbent assay(ELISA),and the relative expression of FIRRE gene was detected by real-time quantitative PCR.PartⅡ:The role of FIRRE in LPS-induced ALI in C57BL/6J mice.Twenty-four healthy and clean SPF grade male C57BL/6J mice(body weight 25-30 g,6-8 weeks old)were randomly divided into 3 groups.The control group was injected with equal volume of normal saline,the ALI group was injected with 5 mg/kg LPS to induce ALI,and the LMWH group was intraperitoneally injected with 5 mg/kg LPS+5 AXa IU/kg LMWH.The changes of hair color,reaction ability,and resistance to external environment were evaluated at 6 h and 12 h after LPS injection.HE staining was used to observe the degree of lung tissue injury,and the lung injury score was calculated.Blood samples were collected from the abdominal main vein,and the expressions of IL-6 and IL-10 in serum were detected by ELISA.The levels of IL-6 and IL-10 in bronchoalveolar lavage fluid(BALF)were detected by ELISA.The relative expression of FIRRE gene was detected by RT-PCR.One-way analysis of variance,LSD-t test,and Dunnett-t test were used.Results Part I:The activity of A549 cells in the LPS group was lower than that in the control group,and the activity of A549 cells in the LPS+LMWH group was higher than that in the LPS group,with a statistically significant difference among multiple groups(F=44.25,P<0.05).The levels of IL-6,IL-10,an

关 键 词：急性肺损伤 脂多糖 FIRRE 低分子肝素钙 细胞损伤 动物实验 

分 类 号：R563[医药卫生—呼吸系统]