海南医科大学学报

目的：结合网络药理学与实验验证，探究吴茱萸汤减轻脓毒症大鼠肺损伤的可能机制。方法：采用TCMSP和ETCM数据库筛选并确定吴茱萸汤中活性成分，利用Uniport数据库挑选出有效靶标，借助GeneCards和OMIM数据库获取脓毒症肺损伤的疾病靶点，使用在线Venny平台选出二者的交集靶点，利用String平台构建PPI网络，运用Cytoscape 3.7.1软件筛选核心靶点，运用DAVID数据库进行GO及KEGG通路富集分析，构建“药物-疾病-通路”网络图。实验验证部分采用CLP法建立脓毒症肺损伤大鼠模型，药物干预后观察各组大鼠生存率及肺组织病理变化，比较肺组织湿/干重比，检测BALF中白细胞计数、相关炎症因子水平及肺组织中NF-κB蛋白表达情况。结果：本研究共筛选得到TNF-α、IL-1β、IL-6等159个吴茱萸汤治疗脓毒症-肺损伤的作用靶点，涉及包括NF-κB signaling pathway在内的20条关键通路。实验验证结果显示：各给药组较模型组大鼠生存率提高（P<0.05），其中吴茱萸汤中剂量组较其余给药组提高最显著（P<0.05）；模型组较假手术组大鼠肺组织湿/干比升高（P<0.05），而各给药组较模型组下降（P<0.05），其中吴茱萸汤中剂量组较其余给药组下降最显著（P<0.05）；模型组较假手术组BALF中白细胞计数及TNF-α、IL-1β、IL-6水平均升高（P<0.05），而各给药组较模型组均降低（P<0.05），其中吴茱萸汤中剂量组较其余给药组降低最显著（P<0.05）；模型组较假手术组大鼠肺组织中NF-κB p65蛋白表达上调（P<0.05），而各给药组较模型组下调（P<0.05），其中吴茱萸汤中剂量组较其余给药组下调最显著（P<0.05）。结论：吴茱萸汤可降低TNF-α、IL-1β、IL-6水平，下调NF-κB蛋白表达，通过抑制脓毒症大鼠炎症以减轻肺损伤，该作用机制可能与NF-κB信号通路有关。

Objective:To pharmacology and experimental verification explore the possible mechanism of Wuzhuyu decoction in reducing lung injury in septic rats based on network. Methods: TCMSP and ETCM databases were used to screen and determine the active ingredients in Wuzhuyu decoction. Effective targets were selected using the Uniport database, and disease targets for sepsis-induced lung injury were obtained using GeneCards and OMIM databases. The intersection targets of the two were selected using the online Venny platform. A PPI network was constructed using the String platform, and core targets were screened using Cytoscape 3.7.1 software.The DAVID database was utilized to analyze GO and KEGG pathway and a "Drug-Disease-Pathway" network diagram was constructed. The experimental validation section used the CLP method to establish a rat model of sepsis-induced lung injury. After drug intervention, the survival rate and pathological changes in lung tissue of each group of rats were observed, and the wet/dry weight ratio of lung tissue was compared. The white blood cell count, levels of related inflammatory factors in BALF and expression of NF-κB protein in lung tissue were detected. Results: This research screened 159 targets of Wuzhuyu decoction for the cure of sepsis lung injury, including TNF-α, IL-1β, IL-6, etc, involving 20 key pathways including NF-κB signaling pathway. The experimental verification results showed that the survival rates of rats in each treatment group were im-proved compared to the model group(P<0.05), while the medium dose group of Wuzhuyu decoction showed the most significant improvement compared to the other treatment groups(P<0.05). The wet/dry ratio of lung tissue in the model group was higher than the sham surgery group(P<0.05), while each treatment group showed a decrease compared to the model group(P<0.05).Among them, the medium dose group of Wuzhuyu decoction showed the most significant decrease compared to the other treatment groups(P<0.05). The model group showed an increase in white blood cell count, TNF-α, IL-1β and IL-6 levels in BALF compared to the sham surgery group(all P<0.05), while each treatment group showed a decrease compared to the model group(all P<0.05). Among them, the medium dose group of Wuzhuyu decoction showed the most significant decrease compared to the other treatment groups(P<0.05). The expression of NF-κB p65 protein in the lung tissue of rats in the model group was upregulated compared to the sham surgery group(P<0.05), while each treatment group was downregulated compared to the model group(P<0.05). Among them, the medium dose group of Wuzhuyu decoction had the most significant downregulation compared to the other treatment groups(P<0.05). Conclusion: Wuzhuyu decoction can reduce the levels of TNF-α, IL-1β and IL-6, downregulate the expression of NF-κB protein, and alleviate lung injury by inhibiting inflammation in septic rats. This mechanism may be related to the NF-κB signaling pathway.