海南医科大学学报2026，Vol.32Issue(8)：587-594,8.DOI:10.13210/j.cnki.jhmu.20260302.001
基于PARP9-DTX3L/Notch轴探讨平喘颗粒抑制树突状细胞成熟的机制研究
Mechanism of Pingchuan Granules in inhibiting dendritic cell maturation via the PARP9-DTX3L/Notch Axis
摘要
Abstract
Objective:To investigate the effects of Pingchuan Granules on dendritic cell maturation mediated by the PARP9-DTX3L/Notch axis,thereby elucidating its molecular mechanism in inhibiting airway inflammation in asthma.Methods:Bone marrow-derived mononuclear cells from C57BL/6J mice were differentiated into dendritic cells using granulocyte-macrophage colony-stimulating factor and interleukin-4,and an in vitro maturation model was established by stimulation with tumor necrosis factor-α.The experiment included the control group,the model group,the Pingchuan Granule group,the inhibitor group,and the inhibitor+Pingchuan Granule group.Cell morphology was observed under an inverted microscope.Flow cytometry was employed to detect dendritic cell purity and the expression of surface molecules CD80,CD86,MHC-Ⅱ,and FcεRI.Western blot and RT-qPCR were used to measure the expression of PARP9,DTX3L,and N1ICD at both protein and mRNA levels,and the nu-cleocytoplasmic translocation of N1ICD was analyzed.Co-immunoprecipitation was performed to detect the PARP9-DTX3L inter-action,N1ICD ubiquitination,and the ADP-ribosylation level of Ub.Results:Compared to the model group,Pingchuan Granule-containing serum significantly inhibited dendritic cell maturation and reduced the expression levels of CD80,CD86,MHC-Ⅱ,and FcεRI(P<0.01).At the molecular level,Pingchuan Granules downregulated the expression of PARP9 and N1ICD proteins,as well as the mRNA levels of PARP9,Hes1,Hes5,and Hey1(P<0.01),and inhibited the nuclear import of N1ICD(P<0.05),whereas they had no significant effect on the protein and mRNA expression of DTX3L as well as the mRNA expression of Notch1(P>0.05).Furthermore,Pingchuan Granules significantly inhibited the interaction between PARP9 and DTX3L,reduced the levels of ADP-ribosylation of Ub,and promoted the ubiquitination of N1ICD,with all differences being statistically significant(P<0.01).Conclusion:Pingchuan Granules can effectively inhibit the maturation of dendritic cells,its mechanism may be related to interfering with the formation of the PARP9-DTX3L complex,thereby suppressing the activation and nuclear translocation of N1ICD in the Notch signaling pathway,and the transcription of its downstream target genes.关键词
平喘颗粒/树突状细胞/Notch信号通路/PARP9-DTX3L复合物/哮喘Key words
Pingchuan Granules/Dendritic cell/Notch signaling pathway/PARP9-DTX3L complex/Asthma分类
医药卫生引用本文复制引用
袁星星,尹兴梅,刘忱阳,王瑶,李超凡..基于PARP9-DTX3L/Notch轴探讨平喘颗粒抑制树突状细胞成熟的机制研究[J].海南医科大学学报,2026,32(8):587-594,8.基金项目
This study was supported by the Young Scientists Fund of the National Natural Science Foundation of China(82205007) （82205007）
Scientific Research Project of Harbin Municipal Science and Technology Bureau(2023ZCZJNS091) 国家自然科学基金青年科学基金(82205007) （2023ZCZJNS091）
哈尔滨市科技局科研项目(2023ZCZJNS091) （2023ZCZJNS091）
