He Shu-Ya, Pan Chun-Shui, Yan Li, Huang Ping, Yan Lu-Lu, Huang Rong, Li Quan, Huo Xin-Mei, Liu Jian, Fan Jing-Yu, Liu Wei, Wang Chao-Ran, Ge Guang-Bo, Sun Kai, Han Jing-Yan
Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, Beijing, China; Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing, China; Key Laboratory of Stasis and Phlegm, State Administration of Traditional Chinese Medicine of the People's Republic of China, Beijing, China; National Key Laboratory of Chinese Medicine Modernization, Beijing, China; Beijing Microvascular Institute of Integration of Chinese and Western Medicine, Beijing, China.
Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing, China; Key Laboratory of Stasis and Phlegm, State Administration of Traditional Chinese Medicine of the People's Republic of China, Beijing, China; National Key Laboratory of Chinese Medicine Modernization, Beijing, China; Beijing Microvascular Institute of Integration of Chinese and Western Medicine, Beijing, China.
Phytomedicine. 2025 Jul 25;143:156839. doi: 10.1016/j.phymed.2025.156839. Epub 2025 May 12.
Acute lung injury (ALI) rapidly progressing into acute respiratory distress syndrome (ARDS) is a major cause of the high fatality rate in acute respiratory infectious diseases. Qing-Fei-Pai-Du-Tang (QFPDT) has a clinical advantage in ALI/ARDS treatment. However, whether QFPDT can improve multiple pathological process involved in pulmonary microcirculatory disturbances during ALI, and the underlying mechanisms remain unclear.
The present study aimed to assess the role of QFPDT in a lipopolysaccharide (LPS)-induced ALI rat model, attempting to disclose the rationale behind the effects of QFPDT.
Male Wistar rats were intraperitoneally injected with LPS (7.5 mg/kg), and received QFPDT (6 g/kg) by gavage either 10 min before (pre-treatment) or 6 hours after (post-treatment) LPS injection. Intravital microscopy, histology, immunohistochemistry and immunofluorescence, flow cytometry, enzyme-linked immunosorbent assay, Western blotting, and proteomics analysis were utilized to investigate the effects and mechanisms of QFPDT. Chemical profiling of QFPDT was performed to identify potential active ingredients.
The results revealed that 6 and 24 hours after LPS injection induced a hyper-inflammatory and hyperpermeability process in rat lung tissues. Pre- and post-treatment with QFPDT attenuated the increase in leukocyte adhesion to pulmonary venules, accompanied by high expression of CD11b and intercellular adhesion molecule-1. Besides, QFPDT attenuated the LPS-induced increase in fluorescein isothiocyanate-dextran leakage from pulmonary microvessels, along with a downregulated expression of junction proteins and an upregulated expression and phosphorylation of Caveolin-1. Moreover, there was a downregulated expression of basement membrane proteins, increased matrix metalloproteinase-9 and cleaved Cathepsin B, and decreased ATP/ADP and ATP/AMP ratios after LPS, all of which were attenuated by QFPDT. Proteomics data evaluated by gene set enrichment analysis, QFPDT pharmacokinetic analysis, combined with molecular docking prediction and surface plasmon resonance validation revealed that QFPDT contained lung-entering prototype ingredients that improved ALI by regulating various key signaling pathway proteins associated with leukocyte adhesion, microvascular hyperpermeability, basement membrane degradation, and oxidative stress.
The present study demonstrates the multifaceted effects of QFPDT and offers insight into better understanding its underlying mechanisms in attenuating LPS-induced pulmonary microcirculatory disturbances and lung dysfunction through a multi-component and multi-target mode, thereby providing evidence supporting the application of QFPDT in ALI/ARDS-related diseases.
急性肺损伤（ALI）迅速进展为急性呼吸窘迫综合征（ARDS）是急性呼吸道传染病高死亡率的主要原因。清肺排毒汤（QFPDT）在ALI/ARDS治疗中具有临床优势。然而，QFPDT是否能改善ALI期间肺微循环障碍所涉及的多个病理过程及其潜在机制仍不清楚。
本研究旨在评估QFPDT在脂多糖（LPS）诱导的ALI大鼠模型中的作用，试图揭示QFPDT作用背后的原理。
雄性Wistar大鼠腹腔注射LPS（7.5 mg/kg），并在LPS注射前10分钟（预处理）或注射后6小时（后处理）通过灌胃给予QFPDT（6 g/kg）。利用活体显微镜检查、组织学、免疫组织化学和免疫荧光、流式细胞术、酶联免疫吸附测定、蛋白质印迹法和蛋白质组学分析来研究QFPDT的作用和机制。对QFPDT进行化学分析以鉴定潜在的活性成分。
结果显示，LPS注射后6小时和24小时在大鼠肺组织中诱导了高炎症和高通透性过程。QFPDT预处理和后处理减轻了白细胞对肺小静脉粘附的增加，伴有CD11b和细胞间粘附分子-1的高表达。此外，QFPDT减轻了LPS诱导的异硫氰酸荧光素-葡聚糖从肺微血管渗漏的增加，同时连接蛋白表达下调，小窝蛋白-1表达和磷酸化上调。此外，LPS后基底膜蛋白表达下调，基质金属蛋白酶-9和半胱天冬酶B裂解增加，ATP/ADP和ATP/AMP比值降低，所有这些均被QFPDT减轻。通过基因集富集分析评估的蛋白质组学数据、QFPDT药代动力学分析，结合分子对接预测和表面等离子体共振验证表明，QFPDT含有进入肺的原型成分，通过调节与白细胞粘附、微血管高通透性、基底膜降解和氧化应激相关的各种关键信号通路蛋白来改善ALI。
本研究证明了QFPDT的多方面作用，并有助于更好地理解其通过多成分、多靶点模式减轻LPS诱导的肺微循环障碍和肺功能障碍的潜在机制，从而为QFPDT在ALI/ARDS相关疾病中的应用提供证据支持。
