Qi Lu, Wang Shun, Guo Tao, Qi Zhuocao, Wu Suwan, Gao Dan, Yan Zhiqiang, Tan Bo, Yang Aidong
The Research Center for Traditional Chinese Medicine, Shanghai Institute of Infectious Diseases and Biosecurity, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, People's Republic of China.
The First College of Clinical Medical Science, China Three Gorges University & Yichang Central People's Hospital, Yichang, 443003, People's Republic of China.
J Inflamm Res. 2024 Dec 21;17:11403-11417. doi: 10.2147/JIR.S498299. eCollection 2024.
Qingdai (QD) is a traditional Chinese medicine (TCM) commonly used in clinical practice to treat acute lung injury/acute respiratory distress syndrome (ALI/ARDS). However, the mechanisms underlying the effects of QD remain not fully understood. This investigation demonstrated QD alleviated LPS-induced ALI in mice and exerted anti-inflammatory effects by inhibiting the JAK2/STAT3 signaling pathway.
The active compounds of QD were identified through UPLC-LTQ-Orbitrap-MS/MS. Network pharmacology predicted potential pharmacological targets and the signaling pathways contributed to the effectiveness of QD in treating ALI. Molecular docking assessed the binding of active components to critical targets. ALI mice triggered by Lipopolysaccharides (LPS) were used for transcriptomic analysis to assess alterations in pulmonary gene expression. The pathological changes of lung tissue were analyzed via HE staining. Proinflammatory cytokine levels in serum were measured using ELISA, and the mRNA expression was measured by RT-qPCR. Western blot analysis evaluated protein expression related to the JAK2/STAT3 signaling pathway. Additionally, RAW264.7 cells induced by LPS were treated with QD to measure proinflammatory cytokines and JAK2/STAT3 signaling pathway protein expression.
Six major components of QD were identified. Network pharmacology predicted JAK2 and STAT3 as targets for QD in ALI treatment, with KEGG analysis highlighting the JAK/STAT signaling pathway. Transcriptomics confirmed the JAK/STAT signaling pathway in the therapeutic effects of QD. Molecular docking demonstrated high binding affinities of bisindigotin, isoindigo, and 6-(3-oxoindolin-2-ylidene)indolo[2,1-b]quinazolin-12-one (IQO) to JAK2 and STAT3. In vivo, QD reduced lung inflammation, downregulated proinflammatory cytokines, and inhibited JAK2/STAT3 signaling pathway. In vitro, QD mitigated LPS-triggered inflammatory responses in RAW264.7 macrophages by inhibiting the same pathway.
The therapeutic effects of QD in ALI might be mediated by the modulation of the JAK2/STAT3 signaling pathway, which may make it a valuable therapeutic strategy for ALI/ARDS.
青黛（QD）是临床常用的一种治疗急性肺损伤/急性呼吸窘迫综合征（ALI/ARDS）的中药。然而，青黛作用的潜在机制仍未完全明确。本研究表明，青黛可减轻脂多糖（LPS）诱导的小鼠ALI，并通过抑制JAK2/STAT3信号通路发挥抗炎作用。
通过超高效液相色谱-线性离子阱-静电场轨道阱串联质谱（UPLC-LTQ-Orbitrap-MS/MS）鉴定青黛的活性成分。网络药理学预测青黛治疗ALI的潜在药理靶点及信号通路。分子对接评估活性成分与关键靶点的结合情况。采用LPS诱导的ALI小鼠进行转录组分析，以评估肺组织基因表达的变化。通过苏木精-伊红（HE）染色分析肺组织的病理变化。采用酶联免疫吸附测定（ELISA）检测血清中促炎细胞因子水平，逆转录-定量聚合酶链反应（RT-qPCR）检测其mRNA表达。蛋白质免疫印迹法（Western blot）分析与JAK2/STAT3信号通路相关的蛋白表达。此外，用青黛处理LPS诱导的RAW264.7细胞，检测促炎细胞因子及JAK2/STAT3信号通路蛋白表达。
鉴定出青黛的6种主要成分。网络药理学预测JAK2和STAT3是青黛治疗ALI的靶点，京都基因与基因组百科全书（KEGG）分析突出了JAK/STAT信号通路。转录组学证实JAK/STAT信号通路在青黛的治疗作用中发挥作用。分子对接显示双吲哚蓝、异靛蓝和6-（3-氧代吲哚啉-2-亚基）吲哚并[2,1-b]喹唑啉-12-酮（IQO）与JAK2和STAT3具有较高的结合亲和力。在体内，青黛减轻肺部炎症，下调促炎细胞因子，并抑制JAK2/STAT3信号通路。在体外，青黛通过抑制相同通路减轻LPS触发的RAW264.7巨噬细胞炎症反应。
青黛对ALI的治疗作用可能是通过调节JAK2/STAT3信号通路介导的，这可能使其成为治疗ALI/ARDS的一种有价值的治疗策略。
