犀角地黄汤合银翘散干预流行性感冒病毒感染的巨噬细胞炎症机制的网络药理学预测及实验验证

邓迪; 马璐瑶; 刘天怡; 周娜; 刘梦华; 杨明锐; 邱宗林; 郝钰

doi:10.3969/j.issn.1006-2157.2023.01.011

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犀角地黄汤合银翘散干预流行性感冒病毒感染的巨噬细胞炎症机制的网络药理学预测及实验验证

实验研究 | 更新时间：2023-02-03

- 犀角地黄汤合银翘散干预流行性感冒病毒感染的巨噬细胞炎症机制的网络药理学预测及实验验证
- Network pharmacological prediction and experimental validation of Xijiao Dihuang Decoction combined with Yinqiao Powder in macrophages infected with influenza virus
- 北京中医药大学学报 2023年46卷第1期页码：57-64
- 作者机构：
  
  北京中医药大学生命科学学院免疫与微生物学系　北京　102488
- 作者简介：
  
  邓迪，女，硕士，医师
  #郝钰，女，博士，教授，博士生导师，主要研究方向：中医药抗感染的免疫机制研究，E-mail：yuhao64@sina.com
- 基金信息：
  
  国家自然科学基金面上项目(81573723)
- DOI：10.3969/j.issn.1006-2157.2023.01.011
  中图分类号： R285.5
- 收稿：2021-12-07，
  
  网络首发：2022-07-22，
  
  纸质出版：2023-01-30
- 稿件说明：
移动端阅览
邓迪, 马璐瑶, 刘天怡, 等. 犀角地黄汤合银翘散干预流行性感冒病毒感染的巨噬细胞炎症机制的网络药理学预测及实验验证[J]. 北京中医药大学学报, 2023,46(1):57-64.

DENG Di, MA Luyao, LIU Tianyi, et al. Network pharmacological prediction and experimental validation of Xijiao Dihuang Decoction combined with Yinqiao Powder in macrophages infected with influenza virus[J]. Journal of Beijing University of Traditional Chinese Medicine, 2023, 46(1): 57-64. DOI: 10.3969/j.issn.1006-2157.2023.01.011.
邓迪, 马璐瑶, 刘天怡, 等. 犀角地黄汤合银翘散干预流行性感冒病毒感染的巨噬细胞炎症机制的网络药理学预测及实验验证[J]. 北京中医药大学学报, 2023,46(1):57-64. DOI： 10.3969/j.issn.1006-2157.2023.01.011.

DENG Di, MA Luyao, LIU Tianyi, et al. Network pharmacological prediction and experimental validation of Xijiao Dihuang Decoction combined with Yinqiao Powder in macrophages infected with influenza virus[J]. Journal of Beijing University of Traditional Chinese Medicine, 2023, 46(1): 57-64. DOI: 10.3969/j.issn.1006-2157.2023.01.011. DOI：

摘要

目的

探究犀角地黄汤合银翘散(XDY)干预流行性感冒病毒(简称“流感病毒”)感染的巨噬细胞炎症的分子机制。

方法

通过GEO数据库公开发表的基因芯片筛选流感病毒PR8感染人巨噬细胞的相关基因；将XDY的活性成分与流感病毒PR8感染人巨噬细胞的相关差异基因的交集输入STRING数据库，获得靶点蛋白相互作用关系，根据蛋白相互作用密切度得到核心靶点，使用Cytoscape3.7.2软件可视化蛋白－蛋白互作网络，对该网络进行拓扑学分析；使用Metascape数据库对靶点分别进行通路富集分析。将巨噬细胞J774A.1分为正常组(正常培养)、模型组(流感病毒PR8感染)、犀角地黄汤合银翘散组(流感病毒PR8感染＋XDY 162.5 mg/L)，干预24 h。实时荧光PCR检测细胞中Toll样受体4(TLR4)、髓样分化因子88(MyD88)、肿瘤坏死因子受体相关分子6(TRAF6)、肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)mRNA水平；免疫荧光检测核转录因子-κB(NF-κB)核转位情况；酶联免疫吸附测定(ELISA)法检测细胞上清液中TNF-α和IL-6水平。

结果

XDY与流感病毒PR8感染人巨噬细胞的差异基因取交集后得到104个共同靶点，通路富集分析结果显示，XDY治疗流感涉及TLR4/NF-κB通路；实时荧光PCR检测结果显示，与模型组比较，犀角地黄汤合银翘散组巨噬细胞的TLR4、MyD88、TRAF6、TNF-α和IL-6 mRNA表达水平降低(

0.01)。免疫荧光显示，与模型组相比，犀角地黄汤合银翘散组巨噬细胞胞内NF-κB与细胞核共定位现象减少，核质比降低(

0.01)。ELISA检测结果显示，与模型组比较，犀角地黄汤合银翘散组细胞上清液中TNF-α和IL-6水平均降低(

0.01)。

结论

犀角地黄汤合银翘散可通过调节TLR4/NF-κB通路减轻流感病毒感染巨噬细胞的炎症反应。

Abstract

Objective

We aimed to explore the anti-inflammatory molecular mechanism of

Xijiao Dihuang

Decoction combined with

Yinqiao

Powder(XDY) in macrophages infected with influenza virus.

Methods

Genes related to human macrophages infected by influenza virus PR8 were screened by the gene chip published in the GEO database.The intersection of the active components of XDY and the related differential genes of human macrophages infected by influenza virus PR8 was input into the STRING database to obtain the interaction relationship between target proteins.The core target was obtained according to the degree of protein interaction.The protein-protein interaction network was visualized with Cytoscape 3.7.2 software

and topology analysis was performed.The Metascape database was used for Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis.Macrophages J774A.1 were divided into the normal group

the model group(influenza virus PR8 infection)

and the XDY group(PR8 + 162.5 mg/L XDY) and treated for 24 h. The mRNA levels of Toll-like receptor 4(TLR4)

myeloid differentiation factor 88(MyD88)

TNF receptor-associated factor 6(TRAF6)

tumor necrosis factor-α(TNF-α)

and interleukin-6(IL-6) were detected by real-time PCR; nuclear factor-κB(NF-κB) translocation was detected by immunofluorescence; and the levels of TNF-α and IL-6 in the supernatant were detected by ELISA.

Results

In total

104 common targets were obtained by analyzing the intersection of the differentially expressed genes in the PR8 and XDY groups.Pathway enrichment analysis showed that the effects of XDY on influenza may be mediated by the TLR4/NF-κB pathway.PCR result showed that compared with the model group

the mRNA levels of TLR4

MyD88

TRAF6

TNF-α

and IL-6 were lower in the XDY group(

0.01).Immunofluorescence result showed that compared with the model group

the nuclear localization of NF-κB in the XDY group was decreased

and the nuclear-cytoplasmic ratio was also decreased(

0.01).ELISA result showed that compared with the model group

the levels of TNF-α and IL-6 in the supernatant in the XDY group were decreased(

0.01).

Conclusion

Xijiao Dihuang

Decoction combined with

Yinqiao

Powder can regulate the TLR4/NF-κB pathway to reduce the inflammatory response in macrophages infected with influenza virus.

关键词

Keywords

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