目的：探索雷公藤甲素是否通过减少胶原诱导关节炎(CIA)小鼠关节腔中性粒细胞浸润来缓解疾病,分析雷公藤甲素是否可以减少类风湿关节炎(RA) 患者中性粒细胞胞外捕获网 (NETs) 形成及其作用机制,为中药雷公藤多苷在RA临床应用提供理论支持。方法：利用网药公共数据库探索雷公藤干预RA疾病的核心靶点;利用DBA1小鼠构建 CIA模型后,给予雷公藤甲素干预60 d,采集小鼠后肢进行 HE染色观察小鼠关节腔炎性细胞浸润程度; 收集RA 患者外周血中性粒细胞,给予雷公藤甲素刺激2 h,通过免疫荧光染色观察NETs形成情况;对RA组和雷公藤甲素治公藤甲素可以缓解CIA 小鼠关节肿胀; 雷公藤甲素还可以抑制RA患者外周血中性粒细胞NETs形成;RA组和雷公藤疗组的中性粒细胞进行转录组测序。结果：通过网络药理学分析,将锁定的重叠基因进行PPI蛋白互作并行KEGG、GO分析,发现IL-17、TNF 信号通路、细胞凋亡相关通路等均与RA疾病密切相关;体内实验表明雷甲素治疗组的转录组测序结果筛选得到389个差异基因,其GO、KEGG通路主要富集为JAK-STAT、TNF 、PI3K-Akt 信号通路等。结论：雷公藤甲素可以通过下调CIA小鼠关节腔中性粒细胞浸润缓解小鼠关节肿胀,同时雷公藤甲素可以减少RA外周血NETs 形成,这一现象可能与JAK-STAT 信号通路、TNF 信号通路、PI3K-Akt等信号通路相关。
Abstract
Objective: To explore whether triptolide can alleviate the disease by reducing the infiltration of neutrophils in the joint cavity of collagen-induced arthritis (CIA) mice, and to analyze whether triptolide can reduce the formation of neutrophil extracellular capture networks (NETs) in patients with rheumatoid arthritis (RA) and its mechanism of action, so as to provide theoretical support for the clinical application of tripterygium glycosides in RA. Methods: The core targets of Tripterygium wilfordii intervention in RA disease were explored by using the public database of network medicine. After the CIA model was constructed using DBA1 mice, triptolide was given for 60 days. The hind limbs of the mice were collected for HE staining and the degree of inflammatory cell infiltration in the joint cavity of the mice was observed. Neutrophils in peripheral blood of RA patients were collected and stimulated with triptolide for 2 h. The formation of NETs was observed by immunofluorescence staining. Treatment with triptolide in RA group and triptolide could alleviate joint swelling in CIA mice; triptolide could also inhibit the formation of NETs in peripheral blood neutrophils of RA patients. Transcriptome sequencing was performed on neutrophils in RA group and Tripterygium wilfordii treatment group. Results: Through network pharmacology analysis, PPI protein interaction was performed on the locked overlapping genes, and KEGG and GO analysis were performed. It was found that IL-17, TNF signaling pathway and apoptosis-related pathway were closely related to RA disease. In vivo experiments showed that 389 differential genes were screened from the transcriptome sequencing results of the triptolide treatment group. The GO and KEGG pathways were mainly enriched in JAK-STAT, TNF, and PI3K-Akt signaling pathways. Conclusion: Triptolide can alleviate joint swelling in CIA mice by down-regulating neutrophil infiltration in the joint cavity of CIA mice, and triptolide can reduce the formation of NETs in peripheral blood of RA.This phenomenon may be related to JAK-STAT signaling pathway, TNF signaling pathway, PI3K-Akt and other signaling pathways.
关键词
类风湿关节炎 /
中性粒细胞 /
雷公藤甲素 /
转录组测序 /
网络药理学
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Key words
Rheumatoid arthritis /
Neutrophils /
Triptolide /
Transcriptome sequencing /
Network pharmacology
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参考文献
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脚注
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基金
国家自然科学基金(82160309)
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