Wang Kun, Fang Haoxiang, Cao Xiaomei
Huixue Research Center, Anhui University of Chinese Medicine, College of Traditional Chinese Medicine, Anhui University of Chinese Medicine, Hefei 230012, China. *Corresponding author, E-mail:
College of Traditional Chinese Medicine, Anhui University of Chinese Medicine, Hefei 230012, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2025 Jul;41(7):585-592.
Objective To observe the effect of mA methylation regulation on Notch1 pathway on the homing of BMSCs in asthma, and the intervention study of traditional Chinese medicine compound Yanghe Pingchuan Granules. Methods Rat bone mesenchymal stem cells(BMSC)and bronchial epithelial cells were cocultured. The extracted cells were divided into: bronchial epithelial cell group, asthma bronchial epithelial cell+mesenchymal stem cell co-culture group (co-culture group), co-culture cell+normal serum group, coculture cell+serum containing optimal drug group, siRNA FTO+normal serum group, siRNA FTO-NC+normal serum group, and siRNA FTO+serum containing optimal drug group. The vitality and cell cycle changes of co-cultured cells were detected. The level and markers of homing BMSC were detected by immunofluorescence staining. The expression of Notch1 pathway related genes were detected by qRT-PCR. The expression of Notch1 pathway related proteins were detected by Western blot. Results Compared with bronchial epithelial cell group, the co-cultured cell group showed an increase in the homing level of BMSCs and the expression of C-X-C motif chemokine receptor 4 (CXCR4), stromal cell-derived factor 1 (SDF-1), Notch1, transcription factor recombination signal binding protein-J (RBP-J), and hairy enhancer of split 1 (Hes1) proteins. Compared with the co-cultured cell group and co-cultured cell+normal serum group, the co-cultured cell+serum containing optimal drug group showed an increase in the homing level of BMSCs and the expressions of CXCR4 and SDF-1, while the protein and mRNA levels of Notch1 and Hes1 decreased. Compared with the siRNA FTO-NC+normal serum group, the siRNA FTO+normal serum group showed an increase in the levels of Notch1, activated Notch1, RBP-J, Hes1 protein, and cell viability, while the level of homing BMSC decreased. Compared with siRNA FTO+normal serum group, the levels of Notch1, RBP-J mRNA, activated Notch1, and Hes1 protein decreased, while the level of homing BMSCs increased in siRNA FTO+serum containing optimal drug group. The levels of Notch1, RBP-J, and Hes1 mRNA were reduced in the co-cultured cells+serum containing optimal drug group. Compared with siRNA FTO+serum containing optimal drug group, the expressions of Notch1, activated Notch1, RBP-J, Hes1 protein and cell viability decreased, while the level of homing BMSCs increased in the co-cultured cells+serum containing optimal drug group. Conclusion Yanghe Pingchuan Granules may promote the homing of BMSCs in asthma and alleviate asthma inflammation by upregulating the expression of FTO and inhibiting the expression of downstream genes in the Notch1 signaling pathway.
目的 观察微小RNA甲基化调控Notch1通路对哮喘中骨髓间充质干细胞归巢的影响，以及中药复方阳和平喘颗粒的干预研究。方法 将大鼠骨髓间充质干细胞（BMSC）与支气管上皮细胞共培养。提取的细胞分为：支气管上皮细胞组、哮喘支气管上皮细胞+间充质干细胞共培养组（共培养组）、共培养细胞+正常血清组、共培养细胞+含最佳药物血清组、siRNA FTO+正常血清组、siRNA FTO-NC+正常血清组、siRNA FTO+含最佳药物血清组。检测共培养细胞的活力和细胞周期变化。通过免疫荧光染色检测归巢BMSC 的水平和标志物。通过qRT-PCR检测Notch1通路相关基因的表达。通过蛋白质印迹法检测Notch1通路相关蛋白的表达。结果 与支气管上皮细胞组相比，共培养细胞组BMSCs的归巢水平以及C-X-C基序趋化因子受体4（CXCR4）、基质细胞衍生因子1（SDF-1）、Notch1、转录因子重组信号结合蛋白-J（RBP-J）和毛状分裂增强子1（Hes1）蛋白的表达增加。与共培养细胞组和共培养细胞+正常血清组相比，共培养细胞+含最佳药物血清组BMSCs的归巢水平以及CXCR4和SDF-1的表达增加，而Notch1和Hes1的蛋白和mRNA水平降低。与siRNA FTO-NC+正常血清组相比，siRNA FTO+正常血清组Notch1、活化Notch1、RBP-J、Hes1蛋白水平和细胞活力增加，而归巢BMSC水平降低。与siRNA FTO+正常血清组相比，siRNA FTO+含最佳药物血清组Notch1、RBP-J mRNA、活化Notch1和Hes1蛋白水平降低，而归巢BMSCs水平增加。共培养细胞+含最佳药物血清组中Notch1、RBP-J和Hes1 mRNA水平降低。与siRNA FTO+含最佳药物血清组相比，共培养细胞+含最佳药物血清组Notch1、活化Notch1、RBP-J、Hes1蛋白表达和细胞活力降低，而归巢BMSCs水平增加。结论 阳和平喘颗粒可能通过上调FTO表达并抑制Notch1信号通路下游基因表达，促进哮喘中BMSCs的归巢并减轻哮喘炎症。
