慢性阻塞性肺疾病模型小鼠中Notch信号通路及上下游调控因子NCSTN和Foxj1的表达  

Expression of Notch signaling pathway NCSTN-Notch-Foxj1 in chronic obstructive pulmonary disease model mice

作　　者：兰茂华 李政録 兰艺 刘楠 胡锦洋 袁国城 顾延会 何志全 Lan Maohua;Li Zhenglu;Lan Yi;Liu Nan;Hu Jinyang;Yuan Guocheng;Gu Yanhui;He Zhiquan(Zunyi Medical University,1.Master’s Degree Students,Class of 2022;Undergraduate Students of Clinical Majors,Class of 2022,Zunyi Guizhou 563006,China;Department of Respiratory Medicine,Affiliated Hospital of Zunyi Medical University,Zunyi Guizhou 563000,China;Laboratory of Morphology,Zunyi Medical University,Zunyi Guizhou 563006,China)

机构地区：[1]遵义医科大学,贵州遵义563006 [2]遵义医科大学附属医院呼吸与危重症医学科一病区科,贵州遵义563000 [3]遵义医科大学形态学实验室,贵州遵义563006

出　　处：《遵义医科大学学报》2025年第2期129-136,共8页Journal of Zunyi Medical University

基　　金：贵州省卫生健康委科学技术基金资助项目(NO:qzwkj2022-035);遵义市科技计划项目[NO:遵市科合HZ字(2022)255];贵州省大学生创新创业训练计划项目(NO:S2024106612279)。

摘　　要：目的检测慢性阻塞性肺疾病(COPD)模型小鼠中气道上皮Notch信号通路、nicastrin(NCSTN)和Foxj1因子的表达。方法选取36只SPF级KM小鼠,随机分为正常对照组(Normal)和COPD组,各组18只,采用香烟烟熏17周建立COPD小鼠模型。使用苏木精-伊红染色(HE)对各组肺组织和气管组织进行病理观察,计算肺泡的平均截距(MLI)、平均肺泡数(MAN)以及支气管和气管的管壁厚度;采用酶联免疫吸附试验(Elisa)检测血清中的肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6);采用免疫组织化学法(IHC)观察各组气管上皮中NCSTN蛋白的定位与表达水平;使用实时荧光定量PCR(RT-qPCR)和免疫印迹(WB)分别测定各组气管上皮NCSTN、Notch1/2/3和Hes1 mRNA和蛋白表达水平,以及Foxj1 mRNA、β-tubulinⅣ蛋白表达水平。结果相较于Normal组,COPD组小鼠肺组织的MLI明显增大(P<0.05)、MAN明显减少(P<0.05)、肺组织支气管和气管管壁厚度明显增加(P<0.05)、气管结构改变;TNF-α和IL-6水平明显增加(P<0.05);NCSTN蛋白在各组气管上皮均有表达,COPD组NCSTN平均吸光度值明显升高(P<0.05);气管上皮中NCSTN、Notch2/3及Hes1 mRNA和蛋白表达水平明显增加(P<0.05)、Foxj1 mRNA和β-tubulinⅣ蛋白表达水平明显降低(P<0.05),Notch1 mRNA和蛋白表达水平差异无统计学意义(P>0.05)。结论提示COPD模型小鼠气道上皮Foxj1的改变与Notch信号通路上游调控基因NCSTN表达异常相关。Objective To detect expression of Notch signaling pathway,Nicastrin(NCSTN)and Foxj1 in chronic obstructive pulmonary disease(COPD)model mice.Methods 36 SPF-grade KM mice were selected and randomly allocated into normal control group(Normal)and COPD group,18 mice in each group.The COPD mouse model was constructed by cigarette smoking method for 17 weeks.Hematoxylin-eosin staining(HE)was implemented to observe the pathological variations of lung and trachea tissues in each group,and the mean linear intercept(MLI),mean alveolar number(MAN),and the thickness of bronchial and tracheal walls were calculated;the enzyme-linked immunosorbent assay(ELISA)was utilized to determine the concentrations of serum tumor necrosis factor-alpha(TNF-α)and interleukin-6(IL-6);immunohistochemistry(IHC)was employed to investigate the protein localization and expression level of NCSTN in the tracheal epithelium of each group;real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting(WB)were employed to determine the mRNA and protein expression levels of NCSTN,Notch1/2/3,Hes1,and ciliated cell-related genes and proteins in the tracheal epithelium of each group,respectively.Results Compared with the Normal group,the lung tissue MLI of mie in the COPD group was significantly larger(P<0.05),MAN was significantly reduced(P<0.05),the thickness of the bronchial and tracheal walls within the lung tissue was significantly augmented(P<0.05),and the structure of the trachea was altered;the levels of TNF-αand IL-6 were significantly increased(P<0.05);and NCSTN proteins were expressed in the tracheal epithelium of all groups.The mean absorbance value of NCSTN was significantly higher in the COPD group(P<0.05);the mRNA and protein expression levels of NCSTN,Notch2/3,and Hes1 in the tracheal epithelium were significantly increased(P<0.05),while the mRNA and protein expression levels of Foxj1 andβ-tubulinⅣwere significantly decreased(P<0.05).However,the mRNA and protein expression levels of Notch1 did not show a significantly difference(P

关 键 词：NCSTN因子 NOTCH信号通路 慢性阻塞性肺疾病 纤毛细胞 

分 类 号：R563.9[医药卫生—呼吸系统]