头孢他啶通过miR-218-5p调控Nfr2/HO-1信号通路对LPS诱导的慢性支气管炎大鼠模型气道炎症和阻塞的改善作用  

Ceftazidime improves airway inflammation and obstruction in LPS-induced chronic bronchitis rat model by regulating Nfr2/HO-1 signaling pathway through miR-218-5p

作　　者：王梦迪 丁浩[1] WANG Mengdi;DING Hao(Department of Respiratory Medicine,Affiliated People's Hospital of Jiangsu University,Zhenjiang 212002,China)

机构地区：[1]江苏大学附属人民医院呼吸内科,镇江212002

出　　处：《免疫学杂志》2024年第8期655-662,共8页Immunological Journal

摘　　要：目的研究头孢他啶通过调控微小RNA-218-5p(miR-218-5p)介导核因子E2-相关因子2(Nfr2)/血红素加氧酶1(HO-1)信号通路对脂多糖(LPS)诱导的慢性支气管炎(CB)大鼠的改善作用。方法在动物水平上,采用气管内注射LPS构建CB模型。将40只大鼠随机分为对照组、模型组、头孢他啶组(肌肉注射头孢他啶200 mg·kg-1)、头孢他啶+miR-218-5p inhibitor组(肌肉注射头孢他啶200 mg·kg-1+尾静脉注射miR-218-5p inhibitor 100 nmol·ml^(-1)),每组10只,各组给药每天2次,连续3 d。苏木精-伊红染色法(HE)检测大鼠肺部组织病理学;实时荧光定量PCR(qRT-PCR)检测miR-218-5p表达;进行气道阻力(RI)检测;酶联免疫吸附法(ELISA)检测支气管肺泡灌洗液(BALF)中炎症介质水平;免疫荧光检测黏蛋白5AC(MUC5AC)、黏蛋白5B(MUC5B)阳性表达;蛋白免疫印迹检测Nfr2/HO-1通路相关蛋白表达。细胞水平上,参照动物水平进行分组,其中头孢他啶组采用10μmol/L头孢他啶处理,头孢他啶+miR-218-5p inhibitor组采用10μmol/L头孢他啶+10μmol/L miR-218-5p inhibitor处理。通过双荧光素酶实验、qRT-PCR、ELISA分别检测各组Nrf2、miR-218-5p、HO-1靶向关系、mRNA表达水平及抗氧化水平。结果动物实验表明,对照组、模型组、头孢他啶组miR-218-5p水平分别为1.00±0.17、0.33±0.06、0.72±0.14;对照组、模型组、头孢他啶组、头孢他啶+miR-218-5p inhibitor组RI分别为(0.23±0.03)、(0.62±0.08)、(0.47±0.06)、(0.59±0.08)cmH_(2)O·s·ml^(-1);IL-6水平分别为(93.84±18.44)(231.16±46.98)(167.89±32.42)、(204.48±40.39)pg·ml^(-1);TNF-α水平分别为(9.06±1.73)、(44.03±8.17)、(31.41±6.72)、(39.15±7.13)pg·ml^(-1);巨噬细胞炎症蛋白1α水平分别为(724.32±133.82)、(1481.18±225.13)、(1007.50±196.69)、(1226.76±207.27)pg·ml^(-1);趋化因子C-C基序配体5水平分别为(166.07±33.61)、(584.20±94.93)、(322.55±57.34)、(463.89±84.04)pg·ml^(-1);MUC5AC阳性率分别为(31.34±5.86)%、(75.48±Objective Research on Cefathiamidine improves chronic bronchitis induced by lipopolysaccharide(LPS)in rats by regulating the microRNA-218-5p(miR-218-5p)mediated Nuclear factor erythroid 2-related factor 2(Nfr2)/Heme oxygenase-1(HO-1)signaling pathway.Methods At the animal level,an LPS-induced CB model was established by intratracheal injection in this study.Forty rats were randomly divided into control group,model group,cefotaxime group(intramuscular injection of cefotaxime 200 mg·kg-1),and cefotaxime+miR-218-5p inhibitor group(intramuscular injection of cefotaxime 200 mg·kg-1+tail vein injection of miR-218-5p inhibitor 100 nmol·ml^(-1)),with 10 rats in each group.Each group received medication twice a day for 3 consecutive days.HE was used to examine the histopathology of rat lung tissues;qRT-PCR was performed to detect miR-218-5p expression;Airway resistance(RI)was measured;ELISA was conducted to measure the levels of inflammatory mediators in bronchoalveolar lavage fluid;Immunofluorescence was used to detect positive expression of Mucoprotein5AC(MUC5AC)and Mucoprotein5B(MUC5B);Western blot was employed to assess the expression of Nrf2/HO-1 pathway-related proteins.At the cellular level,the cells were grouped according to the animal level.The ceftazidime group was treated with 10μmol/L ceftazidime,and the ceftazidime+miR-218-5p inhibitor group was treated with 10μmol/L ceftazidime+10μmol/L miR-218-5p inhibitor.Nrf2,miR-218-5p,HO-1 targeting relationship,mRNA expression level and antioxidant level were detected by double luciferase assay,qRT-PCR and ELISA.Results Animal experiments show that the levels of miR-218-5p in the control group,model group,and cefotaxime group were 1.00±0.17,0.33±0.06,and 0.72±0.14,respectively.The RI in the control group,model group,cefotaxime group,and cefotaxime+miR-218-5p inhibitor group were(0.23±0.03),(0.62±0.08),(0.47±0.06),and(0.59±0.08)cmH_(2)O·s·ml^(-1).IL-6 levels were(93.84±18.44),(231.16±46.98),(167.89±32.42),and(204.48±40.39)pg·ml^(-1),while TNF-αlevel

关 键 词：慢性支气管炎 头孢他啶 微小RNA-218-5p Nfr2/HO-1信号通路 气道炎症 气道黏液高分泌 

分 类 号：R562.21[医药卫生—呼吸系统]