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.
Nan Fang Yi Ke Da Xue Xue Bao. 2024 Dec 20;44(12):2283-2290. doi: 10.12122/j.issn.1673-4254.2024.12.03.
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.
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.
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.
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.
观察miR-139-5p和Notch1信号通路在哮喘大鼠骨髓间充质干细胞（BMSCs）归巢调控中的作用。
将正常大鼠BMSCs与正常或哮喘大鼠的支气管上皮细胞共培养，然后用miR-139-5p模拟物或阴性对照序列进行转染。分析细胞活力和细胞周期的变化，采用免疫荧光染色检测CXCR4和SDF-1的细胞表达。观察转染后BMSC归巢的变化，用RT-qPCR、蛋白质印迹法或ELISA检测Notch1、RBP-J和Hes1 mRNA及蛋白以及Th1/Th2细胞因子的表达。
BMSCs与哮喘支气管上皮细胞共培养显示miR-139-5p、IL-2和IL-12表达显著降低，CXCR4、SDF-1、IL-5、IL-9、Notch1、RBP-J和Hes1表达增加。用miR-139-5p模拟物转染显著增加了共培养细胞中miR-139-5p、IL-2、CXCR4和SDF-1的表达，并降低了IL-5、IL-9、Notch1、活化的Notch1和Hes1的表达水平。相关性分析表明，BMSC归巢与miR-139-5p和IL-12呈正相关，与IL-5表达呈负相关。CXCR4的表达与活化的Notch1呈负相关，SDF-1与miR-139-5p呈正相关，但与Notch1表达呈负相关。
miR-139-5p的高表达通过靶向Notch1信号通路调节Th1/Th2细胞因子的表达，从而促进哮喘中BMSCs的归巢，减轻气道炎症。
