[1]张翠云,李风森,李江涛,等.基于“肺通调水道”理论探究 miRNA-200b-3p通过调节 Notch1/FOXI1轴促进慢性阻塞性肺病发病机制[J].陕西中医,2026,(3):297-303.[doi:DOI:10.3969/j.issn.1000-7369.2026.03.002]
　ZHANG Cuiyun,LI Fengsen,LI Jiangtao,et al.Based on the theory of “lung regulation of waterway”,this study explored the pathogenesis of chronic obstructive pulmonary disease mediated by miRNA-200b-3p mediated Notch1/FOXI1 axis[J].,2026,(3):297-303.[doi:DOI:10.3969/j.issn.1000-7369.2026.03.002]
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基于“肺通调水道”理论探究 miRNA-200b-3p通过调节 Notch1/FOXI1轴促进慢性阻塞性肺病发病机制

《陕西中医》[ISSN:1000-7369/CN:61-1281/TN]

卷:
期数:
2026年3期
页码:
297-303
栏目:
基础研究
出版日期:
2026-03-05

文章信息/Info

Title:
Based on the theory of “lung regulation of waterway”,this study explored the pathogenesis of chronic obstructive pulmonary disease mediated by miRNA-200b-3p mediated Notch1/FOXI1 axis
作者:
张翠云1; 李风森1; 2; 李江涛1; 荆晶1; 2
（1.新疆医科大学第四临床医学院，新疆 乌鲁木齐 830000；2.新疆医科大学附属中医医院 新疆呼吸病研究重点实验室 新疆呼吸系统疾病临床医学研究中心，新疆 乌鲁木齐 830000）
Author(s):
ZHANG Cuiyun1; LI Fengsen1; 2; LI Jiangtao1; JING Jing1; 2
(1.The Fourth Clinical Medicine College of Xinjiang Medical University,Urumqi 830000,China;2.Hospital of Traditional Chinese Medicine Affiliated to Xinjiang Medical University,Xinjiang Key Laboratory of Respiratory Disease Research,Xinjiang Respiratory Obstructive Disease Clinical Medical Research Center,Urumqi 830000,China)
关键词:
慢性阻塞性肺病; miRNA-200b-3p; Notch1/FOXI1轴; 囊性纤维化跨膜电导调节因子; 水通道蛋白; 水液代谢; 生物标志物
Keywords:
Chronic obstructive pulmonary disease; miRNA-200b-3p; Notch1/FOXI1 axis; Cystic fibrosis transmembrane conductance regulator; Aquaporin; Water metabolism; Biological markers
分类号:
R 563.9
DOI:
DOI:10.3969/j.issn.1000-7369.2026.03.002
文献标志码:
A
摘要:
目的：探讨16HBE来源外泌体miR-200b-3p暴露于CSE中的表达，以及miR-200b-3p在Notch1/FOXI1轴中的潜在调节作用。方法：将16HBE细胞经miRNA转染分为空白对照组、CSE干预组、CSE干预+inhibitor NC组、CSE干预+miR-200b-3p-inhibitor组，其中CSE干预16HBE建立慢性阻塞性肺病模型。ELISA法检测各组细胞上清中AQP4、AQP5含量，用qRT-PCR法检测各组细胞上清中Notch1、CFTR、FOXI1、miR-200b-3p mRNA的相对表达，Western blot法检测各组细胞Notch1、FOXI1、CFTR蛋白的相对表达，免疫荧光法检测各组细胞囊性纤维化跨膜电导调节因子（CFTR）荧光强度。外泌体检测：纳米颗粒跟踪分析(NTA)进行表征，Western blot法检测各组细胞外泌体阳性标志物CD9、CD63及各组外泌体内Notch1、FOXI1、CFTR蛋白相对表达水平，qRT-PCR检测miR-200b-3p mRNA的相对表达。结果：与空白对照组比较，CSE干预组AQP4含量升高（P＜0.05），AQP5含量降低（P＜0.05），Notch1、CFTR、FOXI1蛋白及mRNA相对表达降低（P＜0.05），miR-200b-3p mRNA相对表达升高（P＜0.05），CFTR免疫荧光强度降低（P＜0.05）；与CSE干预组比，CSE干预+miR-200b-3p-inhibitor组Notch1、CFTR、FOXI1蛋白及mRNA相对表达升高（P＜0.05），miR-200b-3p mRNA相对表达降低（P＜0.05），CFTR免疫荧光强度增强（P＜0.05）。提取各组上清外泌体，CD9和CD63在各组中的相对表达均为阳性。与空白组对照组相比较，CSE干预组miR-200b-3p mRNA的相对表达升高（P＜0.05），Notch1、FOXI1、CFTR蛋白的相对表达量降低（P＜0.05）。结论：miR-200b-3p通过直接调节其在支气管上皮细胞中的靶基因Notch1/FOXI1在CSE诱导的支气管上皮细胞中AQP4、AQP5失衡发挥关键作用。
Abstract:
Objective:This study investigated the expression of microRNA-200b-3p (miR-200b-3p) derived from bronchial epithelial cells (16HBE) when exposed to cigarette smoke extract (CSE),and explored its potential regulatory role in the Notch1/FOXI1 axis.Methods:16HBE cells were transfected with miRNA to form four experimental groups:control,CSE intervention,CSE intervention+inhibitor NC,and CSE intervention+miR-200b-3p-inhibitor.CSE intervention was used to establish COPD models.ELISA assays detected AQP4 and AQP5 levels in cell supernatants,while qRT-PCR quantified the relative expression of Notch1,CFTR,FOXI1,and miR-200b-3p mRNA.Western blot analysis revealed relative protein expression of Notch1,FOXI1,and CFTR,with immunofluorescence detecting CFTR fluorescence intensity.Exosome characterization was performed using nanoparticle tracking analysis (NTA).Western blot analysis also assessed exosome biomarkers CD9,CD63,and relative expression levels of Notch1,FOXI1,and CFTR proteins,while qRT-PCR monitored the relative expression of miR-200b-3p mRNA.Results:Compared with the blank control group,CSE intervention significantly increased AQP4 content (P<0.05) and decreased AQP5 levels (P<0.05).Notch1,CFTR,and FOXI1 proteins along with their mRNA showed reduced relative expression (P<0.05),while miR-200b-3p mRNA levels rose (P<0.05).CFTR immunofluorescence intensity was also diminished (P<0.05).In contrast,the combined CSE + miR-200b-3p inhibitor treatment elevated Notch1,CFTR,and FOXI1 protein/mRNA expression (P<0.05) while decreasing miR-200b-3p mRNA (P<0.05),with enhanced CFTR immunofluorescence intensity (P<0.05).Extracellular exosomes from all groups tested positive for CD9 and CD63.Compared to the blank control,CSE intervention increased miR-200b-3p mRNA expression (P<0.05) but reduced Notch1,FOXI1,and CFTR protein levels (P<0.05).Conclusion:Our findings demonstrate that miR-200b-3p directly regulates Notch1/FOXI1 in bronchial epithelial cells,playing a pivotal role in addressing AQP4 and AQP5 imbalances induced by CSE.

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备注/Memo

备注/Memo:
国家自然科学基金（地区基金）资助项目（81960848）；新疆维吾尔自治区重点实验室开放课题（2023D04042）；新疆呼吸病研究重点实验室开放课题项目（HXBYJSYS2024008）；新疆医科大学研究生创新创业项目（CXCY2025017）；新疆维吾尔自治区“天山英才”医药卫生高层次人才培养计划（TSYC202301B057）
更新日期/Last Update: 2026-03-05