miR-139-5p调控Notch/RBP-J/Hes1轴促进支气管哮喘骨髓间充质干细胞“归巢”  

MiR-139-5p regulates the Notch/RBP-J/Hes1 axis to promote homing of bone mesenchymal stem cells in bronchial asthma

作　　者：王坤[1,2] 方昊翔 曹晓梅[1] 朱子衡 WANG Kun;FANG Haoxiang;CAO Xiaomei;ZHU Ziheng(College of Traditional Chinese Medicine,Anhui University of Chinese Medicine,Hefei 230031,China;Huixue Research Center,Anhui University of Chinese Medicine,Hefei 230031,China;Graduate School,Anhui University of Chinese Medicine,Hefei 230031,China)

机构地区：[1]安徽中医药大学,中医学院,安徽合肥230031 [2]安徽中医药大学,徽学研究中心,安徽合肥230031 [3]安徽中医药大学,研究生院,安徽合肥230031

出　　处：《南方医科大学学报》2024年第12期2283-2290,共8页Journal of Southern Medical University

基　　金：国家自然科学基金(82205053);安徽省高等学校自然科学研究项目(2022AH050531);新安医学教育部重点实验室开放课题项目(2022xayx03)安徽中医药大学第一批人才支持计划项目(2022rcyb015)。

摘　　要：目的通过将哮喘支气管上皮细胞与骨髓间充质干细胞共培养方式,观察miR-139-5p调控Notch1信号通路对哮喘骨髓间充质干细胞归巢的影响。方法将大鼠骨髓间充质干细胞和支气管上皮细胞共培养,并予miR-139-5p mimics进行干预。实验分为阴性对照组(NC组:正常大鼠骨髓间充质干细胞+支气管上皮细胞共培养)、模型对照组(MC组:正常大鼠骨髓间充质干细胞+哮喘大鼠支气管上皮细胞共培养)、miR-139-5p mimics组(miR-139-5p mimics+MC)、miR-139-5p mimics-NC组(miR-139-5p mimics-NC+MC)。检测细胞活力和细胞周期变化;免疫荧光染色检测CXCR4、SDF-1表达;经细胞转染后观察miR-139-5p表达量和BMSCs归巢水平;免疫印迹观察Notch1/RBP-J/Hes1蛋白表达量;ELISA检测Th1、Th2相关因子表达情况。结果与NC组比较,MC组miR-139-5p、IL-2、IL-12表达量降低(P<0.05),BMSCs归巢水平增加,CXCR4、SDF-1、IL-5、IL-9表达升高(P<0.05),Notch1、RBP-J、Hes1 mRNA、蛋白表达升高(P<0.05)。与MC组、miR-139-5p mimics-NC组比较,miR-139-5p mimics组miR-139-5p、BMSCs归巢水平升高,CXCR4、SDF-1、IL-2表达升高(P<0.05),Notch1、RBP-J、Hes1mRNA、蛋白表达降低,IL-5、IL-9表达降低(P<0.05)。相关性分析显示,BMSCs归巢水平与miR-139-5p、IL-12呈正相关,SDF-1与miR-139-5p呈正相关(P<0.05);与IL-5表达呈负相关,CXCR4与Activated Notch1表达呈负相关,SDF-1与Notch1表达呈负相关(P<0.05)。结论miR-139-5p可能通过靶向Notch1信号通路促进哮喘骨髓间充质干细胞归巢,作用于Th1、Th2细胞因子表达,改善哮喘气道炎症。Objective To observe the role of miR-139-5p and Notch1 signaling pathway in regulation of homing of bone mesenchymal stem cells(BMSCs)of asthmatic rats.Methods Normal rat BMSCs were co-cultured with bronchial epithelial cells from normal or asthmatic rats,followed by transfection with miR-139-5p mimics or a negative control sequence.The changes in cell viability and cell cycle were analyzed,and the cellular expressions of CXCR4 and SDF-1 were detected using immunofluorescence staining.The changes of BMSC homing after the transfection were observed,and the expressions of Notch1,RBP-J,and Hes1 mRNAs and proteins and Th1/Th2 cytokines were detected with RT-qPCR,Western blotting or ELISA.Results The co-cultures of BMSCs and asthmatic bronchial epithelial cells showed significantly decreased expressions of miR-139-5p,IL-2 and IL-12 and increased expressions of CXCR4,SDF-1,IL-5,IL-9,Notch1,RBP-J,and Hes1.Transfection with miR-139-5p mimics significantly increased the expressions of miR-139-5p,IL-2,CXCR4 and SDF-1 and lowered the expression levels of IL-5,IL-9,Notch1,activated Notch1,and Hes1 in the co-cultured cells.Correlation analysis showed that BMSC homing was positively correlated with miR-139-5p and IL-12 and negatively correlated with IL-5 expression.The expression of CXCR4 was negatively correlated with activated Notch1,and SDF-1 was positively correlated with miR-139-5p but negatively correlated with Notch1 expression.Conclusion High expression of miR-139-5p promotes homing of BMSCs in asthma by targeting the Notch1 signaling pathway to regulate the expressions of Th1/Th2 cytokines,thereby alleviating airway inflammation.

关 键 词：支气管哮喘 骨髓间充质干细胞 miR-139-5p Notch1信号通路 

分 类 号：R562.25[医药卫生—呼吸系统]