浙江中医药大学学报2025，Vol.49Issue(12)：1493-1505,13.DOI:10.16466/j.issn1005-5509.2025.12.001
基于天然免疫记忆效应探究解毒祛瘀滋阴方通过抑制JAK-STAT信号通路治疗系统性红斑狼疮的机制
Explore the Mechanism of Jiedu Quyu Ziyin Prescription in Treating SLE by Inhibiting the JAK-STAT Signaling Pathway Based on Innate Immune Memory
摘要
Abstract
[Objective]To explore the intrinsic connection between the pathogenesis of systemic lupus erythematosus(SLE)and the innate immune memory molecules on Janus kinase-signal transducer and activator of transcription(JAK-STAT)signaling pathway by using transcriptomics,and to study the potential mechanism of action of Jiedu Quyu Ziyin Prescription(JP).[Methods]Herbal drug-containing serum was prepared and screened for optimal concentration by CCK-8 method.Bone marrow macrophages(BMDM)from MRL/lpr mice were cultured in vitro and treated with JP-containing serum,lipopolysaccharide(LPS)and interferon-γ(IFN-γ).The cells were randomly divided into blank group(A group),primary stimulation group(LPS,B group),secondary stimulation group(IFN-γ+LPS,C group),primary stimulation+JP group(LPS+JP,D group)and secondary stimulation+JP group(IFN-γ+LPS+JP,E group).Subsequently,RNA next-generation sequencing was applied to screen out the differently expressed genes,which were validated with bioassay and reverse transcription-quantitative polymerase chain reaction(RT-qPCR).Knockdown of histone H3.3 by small interfering RNA(siRNA)transfection was used to verify the mRNA expression of related genes.[Results]The JP-containing serum with a volume fraction of 2.5%had a promotional effect on the proliferation of BMDM.Compared with group A,some differentially expressed genes(DEGs)in groups B and C were associated with histone modifications.The jointly up-regulated DEGs in the three datasets were enriched in immune response,intrinsic immune response and JAK-STAT signaling pathway.Four key genes including signal transducer and activator of transcription 1(STAT1),interferon-stimulated gene 15(ISG15),ubiquitin-specific peptidase 18(USP18),interferon-induced protein with tetratricopeptide repeats 1(IFIT1)located in JAK-STAT signaling pathway were screened by the protein-protein interaction(PPI)network,and RT-qPCR validation was consistent with the transcriptome results.The expression of interferon-stimulated genes(ISGs)on JAK-STAT signaling pathway was down-regulated after the action of JP,and the down-regulated DEGs were enriched in defense response,response to IFN-γ and JAK-STAT signaling pathway.After knockdown of histone H3.3,the related genes were no longer up-regulated after re-stimulation and lost their memory capacity.[Conclusion]The activation of innate immune memory molecules in JAK-STAT signaling pathway is closely related to the pathogenesis of SLE,and the deposition of histone H3.3 helps to accelerate the formation of memory.JP can down-regulate the transcriptional level of the relevant molecules in JAK-STAT signaling pathway,reduce the innate immune memory ability,and thus achieve the goal of treating SLE.关键词
JAK-STAT信号通路/解毒祛瘀滋阴方/天然免疫记忆效应/系统性红斑狼疮/转录组学/MRL/lpr狼疮小鼠/骨髓巨噬细胞/干扰素刺激基因Key words
JAK-STAT signaling pathway/Jiedu Quyu Ziyin Prescription/innate immune memory effect/systemic lupus erythematosus/transcriptome/MRL/lpr lupus mice/bone marrow-derived macrophage/interferon-stimulated gene分类
医药卫生引用本文复制引用
王丹嫣,陈可玥,刘伊苁,金越,李荣群..基于天然免疫记忆效应探究解毒祛瘀滋阴方通过抑制JAK-STAT信号通路治疗系统性红斑狼疮的机制[J].浙江中医药大学学报,2025,49(12):1493-1505,13.基金项目
浙江省自然科学基金项目(LTGY24H270003) （LTGY24H270003）
浙江省中医药科技计划项目(2023ZF014) （2023ZF014）
第五批全国中医临床优秀人才研修项目(国中医药人教函[2022]239号) Zhejiang Province Natural Science Foundation Project(LTGY24H270003) （国中医药人教函[2022]239号）
Zhejiang Province Traditional Chinese Medicine Technology Plan Project(2023ZF014) （2023ZF014）
The Fifth Batch of National TCM Clinical Excellent Talents Training Project(G.TCM.R.J.H.[2022]239) （G.TCM.R.J.H.[2022]239）
