目的 基于网络药理学及体外实验探讨柴胡救肺方治疗 COVID-19 的作用机制。方法 （1）利用 TCMSP、BATMAN-TCM、TCMIP 数据库获取柴胡救肺方活性成分及其作用靶点；通过 GEO 数据库检索并下载 与 COVID-19 感染相关的 GSE152418 基因表达谱，筛选出差异表达基因即为 COVID-19 疾病相关靶点；对 COVID-19 的差异表达基因进行免疫浸润分析。对柴胡救肺方作用靶点与 COVID-19 疾病相关靶点取交集，所 得交集基因即为柴胡救肺方治疗 COVID-19 的潜在作用靶点；通过 STRING 数据库构建潜在作用靶点 PPI 网 络，并筛选柴胡救肺方治疗 COVID-19 的核心靶点Ⅰ；对潜在作用靶点进行最小绝对收缩和选择算子（LASSO） 回归分析，得到柴胡救肺方治疗 COVID-19 的核心靶点Ⅱ；对核心靶点Ⅰ与核心靶点Ⅱ取交集，所得基因即为 柴胡救肺方治疗 COVID-19 的关键靶点。（2）采用 500 ng·mL-1 脂多糖（LPS）诱导嗜酸性粒细胞复制体外炎症细 胞模型，并用柴胡救肺方含药血清低、中、高剂量（5%、10%、15% 含药血清）以及 5 μmol·L-1 BAY 11-7082 （NF-κB p65 抑制剂）、10% 柴胡救肺方含药血清+BAY 11-7082 进行干预。采用 RT-PCR 法检测细胞中 IL-10、 IL-1β、IL-1ɑ、CCL11、CCL24、CCL26 mRNA 表达水平；ELISA 法检测细胞上清液中 IL-10、IL-1β、IL-1ɑ、 CCL11、CCL24、CCL26 的水平；Western Blot 法检测细胞中 p-NF-κB p65/NF-κB p65、p-STAT3/STAT3 蛋白 表达水平。结果 （1）共获得柴胡救肺方活性成分 148 个，活性成分作用靶点 270 个，COVID-19 相关差异表 达基因（COVID-19 疾病相关靶点）4 189 个。对柴胡救肺方活性成分作用靶点与 COVID-19 疾病相关靶点取交 集，得到柴胡救肺方治疗 COVID-19 的潜在作用靶点（共同靶点）67 个，筛选出 10 个核心靶点Ⅰ。通过 LASSO 回归分析筛选出了 15 个核心靶点Ⅱ；对核心靶点Ⅰ与核心靶点Ⅱ取交集，共获得 3 个关键靶点：IL10、IL1B、 EGFR。免疫浸润分析发现，CD4+ T 细胞记忆亚群和嗜酸性粒细胞在 COVID-19 患者中低表达（P＜0.05，P＜0.01）， 浆细胞在 COVID-19 患者中高表达（P＜0.001）；IL10 在浆细胞、单核细胞和嗜酸性粒细胞中高表达，IL1B 在 嗜酸性粒细胞中高表达，EGFR 在浆细胞中高表达。（2）与对照组比较，模型组细胞 IL-10 mRNA 表达显著下调 （P＜0.01），IL-1β、IL-1ɑ、CCL11、CCL24、CCL26 mRNA 表达显著上调（P＜0.01，P＜0.001）；细胞上清液 中的 IL-10 含量显著降低（P＜0.01），IL-1β、IL-1ɑ、CCL11、CCL24、CCL26 含量显著升高（P＜0.05，P＜ 0.01，P＜0.001）；细胞的 p-NF-κB p65/NF-κB p65、p-STAT3/STAT3 蛋白表达比值均显著升高（P＜0.001）。 与模型组比较，柴胡救肺方含药血清低、中、高剂量组细胞 IL-10 mRNA 表达显著上调（P＜0.05，P＜0.01）， IL-1β、IL-1ɑ、CCL11、CCL24、CCL26 mRNA 表达水平显著降低（P＜0.01，P＜0.001）；柴胡救肺方含药血清低剂量组细胞上清液中的 IL-1β 含量明显降低（P＜0.05）；柴胡救肺方含药血清中剂量组细胞上清液中的 IL-10 含量明显升高（P＜0.05），IL-1β、CCL24、CCL26 含量显著下降（P＜0.05，P＜0.001）；柴胡救肺方含药血清高 剂量组细胞上清液中的 IL-10 含量显著升高（P＜0.01），IL-1β、IL-1ɑ、CCL11、CCL24、CCL26 的含量显著下 降（P＜0.05，P＜0.01，P＜0.001）；柴胡救肺方含药血清低剂量组细胞的 p-STAT3/STAT3 蛋白表达比值显著降 低（P＜0.001），柴胡救肺方含药血清中、高剂量组细胞的 p-NF-κB p65/NF-κB p65、p-STAT3/STAT3 蛋白表 达比值均显著降低（P＜0.01，P＜0.001）；柴胡救肺方含药血清组（10%）、BAY 11-7082 组、柴胡救肺方含药 血清（10%）+BAY 11-7082 组细胞的 p-STAT3/STAT3 蛋白表达比值及 IL-1β 水平均显著降低（P＜0.01，P＜ 0.001）。结论 柴胡救肺方可能通过作用于 NF-κB/STAT3 信号通路的磷酸化，抑制 IL-1β 的合成，从而发挥 抗 COVID-19 的作用。

Objective To investigate the mechanism of Chaihu Jiufei Decoction （CHJFF） in treating COVID-19 using network pharmacology and in vitro experiments. Methods（1） Active components of CHJFF and their targets were retrieved from TCMSP， BATMAN-TCM， and TCMIP databases. The GSE152418 gene expression profile related to COVID-19 infection was downloaded from the GEO database， and differentially expressed genes （DEGs） were identified as COVID-19-related targets. Immune infiltration analysis was performed on COVID-19 DEGs. The intersection between CHJFF targets and COVID-19-related targets yielded potential therapeutic targets of CHJFF for COVID-19. A protein-protein interaction （PPI） network of potential targets was constructed using the STRING database，and core targets I for CHJFF in treating COVID-19 were screened. Least absolute shrinkage and selection operator （LASSO） regression analysis was applied to identify core targets II. The intersection of core targets I and II yielded key targets of CHJFF for COVID-19.（2） An in vitro inflammatory cell model was established by inducing eosinophils with 500 ng·mL-1 LPS and treating them with low-，medium-，and high-dose CHJFF-containing serum （5%，10%，15%），5 μmol·L-1 BAY 11-7082（NF-κB p65 inhibitor），or 10% CHJFF-containing serum + BAY 11-7082. RT-PCR was used to measure mRNA expression levels of IL-10，IL-1β，IL-1α，CCL11，CCL24，and CCL26 in cells. ELISA was performed to detect levels of IL-10，IL-1β，IL-1α，CCL11，CCL24，and CCL26 in cell supernatants. Western Blot was used to assess protein expression ratios of p-NF- κB p65/NF- κB p65 and p-STAT3/ STAT3. Results （1） A total of 148 active components of CHJFF，270 component-related targets，and 4 189 COVID- 19-related DEGs were identified. The intersection analysis revealed 67 potential therapeutic targets （common targets） of CHJFF for COVID-19，from which 10 core targets I were screened. LASSO regression identified 15 core targets II. The intersection of core targets I and II yielded three key targets：IL10，IL1B，and EGFR. Immune infiltration analysis showed that memory CD4+ T cells and eosinophils were downregulated in COVID-19 patients （P＜0.05，P＜0.01）， while plasma cells were upregulated （P＜0.001）. IL10 was highly expressed in plasma cells， monocytes， and eosinophils；IL1B was upregulated in eosinophils；and EGFR was overexpressed in plasma cells.（2） Compared with the control group， the model group showed significantly downregulated IL-10 mRNA expression （P＜0.01）， significantly upregulated mRNA expression of IL-1β，IL-1α，CCL11，CCL24，and CCL26（P＜0.01，P＜0.001）； significantly decreased IL-10 content in cell supernatant （P＜0.01），significantly increased IL-1β，IL-1α，CCL11， CCL24，and CCL26 content （P＜0.05，P＜0.01，P＜0.001）；and significantly elevated protein expression ratios of p-NF-κB p65/NF-κB p65 and p-STAT3/STAT3（P＜0.001）. Compared with the model group，the low-，medium-， and high-dose CHJFF-containing serum groups exhibited significantly upregulated IL-10 mRNA expression （P＜0.05， P＜0.01）， significantly downregulated mRNA expression levels of IL-1β， IL-1α， CCL11， CCL24， and CCL26 （P＜0.01，P＜0.001）；the low-dose CHJFF-containing serum group showed significantly reduced IL-1β content in cell supernatant （P＜0.05）； the medium-dose CHJFF-containing serum group demonstrated significantly increased IL-10 content （P＜0.05） and significantly decreased IL-1β，CCL24，and CCL26 content （P＜0.05，P＜0.001） in cell supernatant；the high-dose CHJFF-containing serum group displayed significantly elevated IL-10 content （P＜ 0.01） and significantly reduced IL-1β， IL-1α， CCL11， CCL24， and CCL26 content （P＜0.05， P＜0.01， P＜ 0.001） in cell supernatant； the low-dose CHJFF-containing serum group showed significantly decreased p-STAT3/ STAT3 protein expression ratio （P＜0.001）， while the medium- and high-dose CHJFF-containing serum groups exhibited significantly reduced protein expression ratios of p-NF-κB p65/NF-κB p65 and p-STAT3/STAT3（P＜0.01， P＜0.001）；the CHJFF-containing serum （10%） group，BAY 11-7082 group，and CHJFF-containing serum （10%） + BAY 11-7082 group all showed significantly decreased p-STAT3/STAT3 protein expression ratios and IL-1β levels （P＜0.01，P＜0.001）. Conclusion CHJFF may exert anti-COVID-19 effects by inhibiting IL-1β synthesis through suppression of NF-κB/STAT3 signaling pathway phosphorylation.

R285.5

国家中医药管理局科研项目（2023ZYLCYJ02-21）；国家中医药管理局中医药创新团队及人才支持计划项目（ZYYCXTD-D-202203）；广东省 重点领域研发计划项目（2023B1111020003）；广东省中医急症研究重点实验室开放课题（KF2023JZ06）；广州市科技局市校院联合资助项目 （广州市中西医结合防治新发突发传染病重点实验室，202201020382）；广东省中医院朝阳人才科研专项（ZY2022KY10，ZY2022YL04）。