槲皮素对家兔气管狭窄形成过程的影响及其机制研究  

Effects and mechanism of quercetin on the formation of tracheal stenosis in rabbits

作　　者：覃财成 周磊 黎雨[1] 李文涛[1] 徐明鹏 李殷康 柳广南[1] Qin Caicheng;Zhou Lei;Li Yu;Li Wentao;Xu;Mingpeng;Li Yinkang;Liu Guangnan(Department of Respiratory and Critical Care Medicine The second Affiliated Hospital of Guangxi Medical University,Nanning 530007,China;Hunan Chest Hospital,Changsha 410200,China)

机构地区：[1]广西医科大学第二附属医院呼吸与危重症医学科,南宁530007 [2]湖南省胸科医院,长沙410200

出　　处：《广西医科大学学报》2020年第5期799-804,共6页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81760001)。

摘　　要：目的:探讨槲皮素干预良性气道狭窄的可能机制及SIRT1在其中的调控作用。方法:将18只家兔随机分为空白对照组、模型组、槲皮素组。除空白对照组外,其余组建立家兔气管狭窄模型。术后1~10 d,空白对照组不予特殊处理,模型组予以生理盐水灌胃(120 mg/kg,1次/d),槲皮素组予槲皮素灌胃(120 mg/kg,1次/d)。术后第11天将家兔安乐死,收集气管组织标本。运用实时荧光定量PCR(qPCR)检测白细胞介素6(IL-6)mRNA、沉默信息调节因子2相关酶1(SIRT1)mRNA的相对表达量;运用免疫组化检测SIRT1蛋白表达情况。在细胞实验部分,予脂多糖(LPS)刺激人胚肺成纤维细胞(WI-38),设置为空白对照组、模型组(LPS 100 ng/L)、槲皮素低剂量组(LPS 100 ng/L+槲皮素5μmol/L)、槲皮素中剂量组(LPS 100 ng/L+槲皮素10μmol/L)、槲皮素高剂量组(LPS 100 ng/L+槲皮素20μmol/L)。运用qPCR检测IL-6 mRNA、SIRT1 mRNA的相对表达量;运用Western blotting检测SIRT1蛋白表达情况。结果:动物实验中,模型组气管狭窄度大于空白对照组;与模型组对比,槲皮素组狭窄度减少;与空白对照组对比,模型组气管黏膜组织中SIRT1 mRNA和蛋白的表达下调,IL-6 mRNA表达上调;与模型组对比,槲皮素组SIRT1 mRNA和蛋白表达上调,IL-6 mRNA表达下调(均P<0.05)。细胞实验中,与空白对照组对比,模型组IL-6 mRNA表达上调,SIRT1 mRNA和蛋白表达下调;与模型组对比,槲皮素低、中、高剂量组IL-6 mRNA表达下调,SIRT1mRNA和蛋白表达上调(均P<0.05);槲皮素低、中、高剂量组之间两两比较,IL-6 mRNA的表达差异有统计学意义(均P<0.05);槲皮素中剂量组对SIRT1 mRNA和蛋白表达上调效应最明显,与低剂量组和高剂量组比较,均有统计学差异(均P<0.05)。结论:槲皮素可能通过上调SIRT1抑制炎症因子IL-6的表达,减轻炎症反应,从而改善家兔气管损伤后狭窄。Objective:To explore the potential mechanism of quercetin in the intervention of benign airway stenosis and the regulatory role of SIRT1 in it.Methods:Eighteen rabbits were randomly divided into blank control group,model group,and quercetin group.Except for the rabbits in the blank control group,the tracheal stenosis models of rabbits were established in the other groups.From 1 d to 10 d after modeling,the blank control group received no treatments,the model group was given saline(120 mg/kg,1 time/day)and quercetin group was given quercetin(120 mg/kg,1 time/day)by gavage.The rabbits were euthanized at 11 d after treatment,and tracheal tissue samples were collected.The relative mRNA expressions of interleukin 6(IL-6)and silencing information regulator 2 related enzyme 1(SIRT1)were detected by real-time fluorescence quantitative PCR(qPCR).The expression of SIRT1 protein was detected by immunohistochemistry.In the part of cell experiment,human embryonic lung fibroblasts(WI-38)were stimulated by lipopolysaccharide(LPS)then divided into blank control group,model group(LPS 100 ng/L),low dose quercetin group(LPS 100 ng/L+quercetin 5μmol/L),middle dose quercetin group(LPS 100 ng/L+quercetin 10μmol/L),and high dose quercetin group(LPS 100 ng/L+quercetin 20μmol/L).The relative mRNA expression of IL-6 and SIRT1 were detected by qPCR,and the expression of SIRT1 protein was detected by Western blotting.Results:In the animal experiment,the degree of tracheal stenosis in the model group was larger than that in the blank control group,and it was significant decreased in the quercetin group compared with the model group.Compared with the blank control group,the expression of SIRT1 mRNA and protein were down-regulated,while the expression of IL-6 mRNA was up-regulated in the tracheal mucosa of the model group.Compared with the model group,the expression of SIRT1 mRNA and protein were up-regulated and the expression of IL-6 mRNA was down-regulated in the quercetin group(all P<0.05).In the cell experiment,compared with the blank

关 键 词：良性气管狭窄 槲皮素 SIRT1 IL-6 

分 类 号：R562[医药卫生—呼吸系统]