Li Yujia, Li Bin, Wang Pan, Wang Qinghua
Institute of Blood Transfusion, Chinese Academy of Medical Sciences and Peking Union Medical College, Chengdu, China.
The Joint Laboratory on Transfusion-transmitted Diseases (TTD) Between Institute of Blood Transfusion, Chinese Academy of Medical Sciences and Nanning Blood Center, Nanning, China.
Front Pharmacol. 2021 Oct 26;12:722126. doi: 10.3389/fphar.2021.722126. eCollection 2021.
Qingfei Paidu decoction (QPD) and Xuanfei Baidu decoction (XBD) are two typical traditional Chinese medicines with proven efficacy for the treatment of SARS-CoV-2, although the underlying mechanism is not well defined. Blunted immune response and enhanced production of pro-inflammatory cytokines (cytokine storm) are two main features observed in patients infected with SARS-CoV-2. Analysis based on network pharmacology has revealed that both QPD and XBD played an important role in the regulation of host immunity. We therefore investigated the role of QPD and XBD in the modulation of innate immunity , focusing on the type 1 interferon (IFN) signaling pathway in A549 cells and pro-inflammatory cytokine production in macrophages. A549 cells were treated with QPD or XBD and the production of endogenous IFNα and IFNβ as well as the expression levels of some interferon-stimulated genes (ISGs) were detected by reverse transcriptase-quantitative PCR (RT-qPCR). Macrophages derived from THP-1 cells were treated with QPD or XBD and their pro-inflammatory cytokine expression levels were measured by RT-qPCR, 6 h post LPS stimulation. In addition, the expression levels of some pro-inflammatory cytokines were further analyzed by ELISA. The effect of QPD and XBD on the NF-κB signaling pathway and the pinocytosis activity of THP-1-derived macrophages were evaluated by Western blot and neutral red uptake assay, respectively. Although QPD and XBD showed very little effect on the type 1 IFN signaling pathway in A549 cells, either QPD or XBD markedly inhibited the production of pro-inflammatory markers including interleukin-6, tumor necrosis factor-α, monocyte chemotactic protein-1, and chemokine ligand 10 in THP-1-derived M1 macrophages. In addition, the phosphorylation of IκBα and NF-κB p65 during the process of macrophage polarization was significantly suppressed following QPD or XBD treatment. QPD and XBD also suppressed the pinocytosis activity of macrophages. QPD and XBD have been shown to have robust anti-inflammatory activities . Our study demonstrated that both QPD and XBD decreased pro-inflammatory cytokine expression, inhibited the activation of the NF-κB signaling pathway, and blunted pinocytosis activity in THP-1-derived macrophages.
清肺排毒汤（QPD）和宣肺败毒汤（XBD）是两种经证实对治疗新型冠状病毒肺炎有效的典型中药，尽管其潜在机制尚不完全清楚。免疫反应减弱和促炎细胞因子（细胞因子风暴）产生增加是新型冠状病毒肺炎患者的两个主要特征。基于网络药理学的分析表明，QPD和XBD在调节宿主免疫方面均发挥重要作用。因此，我们研究了QPD和XBD在调节固有免疫中的作用，重点关注A549细胞中的1型干扰素（IFN）信号通路以及巨噬细胞中促炎细胞因子的产生。用QPD或XBD处理A549细胞，通过逆转录定量PCR（RT-qPCR）检测内源性IFNα和IFNβ的产生以及一些干扰素刺激基因（ISG）的表达水平。用QPD或XBD处理源自THP-1细胞的巨噬细胞，在LPS刺激6小时后，通过RT-qPCR测量其促炎细胞因子表达水平。此外，通过酶联免疫吸附测定（ELISA）进一步分析一些促炎细胞因子的表达水平。分别通过蛋白质免疫印迹法和中性红摄取试验评估QPD和XBD对NF-κB信号通路的影响以及THP-1衍生巨噬细胞的胞饮活性。尽管QPD和XBD对A549细胞中的1型IFN信号通路影响很小，但QPD或XBD均显著抑制源自THP-1的M1巨噬细胞中促炎标志物的产生，包括白细胞介素-6、肿瘤坏死因子-α、单核细胞趋化蛋白-1和趋化因子配体10。此外，在QPD或XBD处理后，巨噬细胞极化过程中IκBα和NF-κB p65的磷酸化被显著抑制。QPD和XBD还抑制了巨噬细胞的胞饮活性。QPD和XBD已被证明具有强大的抗炎活性。我们的研究表明，QPD和XBD均降低了促炎细胞因子的表达，抑制了NF-κB信号通路的激活，并减弱了THP-1衍生巨噬细胞的胞饮活性。
