Key Laboratory of Computational Chemistry-Based Natural Antitumor Drug Research & Development, Liaoning Province, Engineering Research Center of Natural Medicine Active Molecule Research & Development, Liaoning Province, Key Laboratory of Natural Bioactive Compounds Discovery & Modification, Shenyang; School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, Shenyang, Liaoning 110016, China.
Key Laboratory of Computational Chemistry-Based Natural Antitumor Drug Research & Development, Liaoning Province, Engineering Research Center of Natural Medicine Active Molecule Research & Development, Liaoning Province, Key Laboratory of Natural Bioactive Compounds Discovery & Modification, Shenyang; School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, Shenyang, Liaoning 110016, China.
Phytomedicine. 2023 Jun;114:154744. doi: 10.1016/j.phymed.2023.154744. Epub 2023 Mar 6.
Acute lung injury (ALI) has the attribution of excessive inflammation of the lung. Jinzhen oral liquid (JO), a famous Chinese recipe used to treat ALI, has a favorable therapeutic effect on ALI. However, its anti-inflammatory mechanism has not been extensively studied.
This study was to elucidate the effects of JO on lipopolysaccharide (LPS)-induced ALI and its molecular mechanism.
An ALI model was established by intratracheal instillation of LPS (2 mg/50 μl). The open field experiment was carried out to explore the spontaneous movement and exploratory behavior of ALI mice. Cytokines levels concentrations (IL-6, IL-10 and TNF-α) were determined by enzyme-linked immunosorbent assay (ELISA). Network pharmacology was used to predict the mechanism of JO against ALI. Immunofluorescence, co-immunoprecipitation, fluorescence resonance energy transfer (FRET), Western blot and RT-PCR were used to verify the molecular mechanisms of JO.
The in vivo results suggested that JO (1, 2, 4 g/kg) dose-dependently improved the exercise performance of mice and reduced the lung W/D weight ratio as well as the production of IL-6 and TNF-α, but increased the release of IL-10 in the ALI group. The network pharmacological analysis demonstrated that the Toll-like receptor (TLR) pathway might be the fundamental action mechanisms of JO against ALI. Immunofluorescence staining and co-immunoprecipitation analysis showed that JO decreased the expression levels of TLR4 and MyD88 and reduced their interaction in the lung tissue of ALI mice. Meanwhile, JO decreased nuclear translocation and phosphorylation of NF-κB P65. The results from cellular experiments were in line with those in vivo. The FRET experiment also confirmed that JO disturbed the interaction of TLR4 and MyD88. Subsequently, we also found that the six indicative components of JO have the similar therapeutic effect as JO.
In summary, we suggested that JO suppressed the TLR4/MyD88/NF-κB signaling pathway, thus inhibiting LPS-induced ALI in vitro and in vivo. The clarified mechanism provided an important theoretical basis and a novel treatment strategy for the ALI treatment of JO.
急性肺损伤（ALI）的特征是肺部过度炎症。金珍口服液（JO）是一种用于治疗 ALI 的著名中药方剂，对 ALI 具有良好的治疗效果。然而，其抗炎机制尚未得到广泛研究。
本研究旨在阐明 JO 对脂多糖（LPS）诱导的 ALI 的作用及其分子机制。
通过气管内滴注 LPS（2mg/50μl）建立 ALI 模型。通过旷场实验探讨 ALI 小鼠的自发运动和探索行为。酶联免疫吸附试验（ELISA）测定细胞因子浓度（IL-6、IL-10 和 TNF-α）。采用网络药理学预测 JO 治疗 ALI 的作用机制。免疫荧光、免疫共沉淀、荧光共振能量转移（FRET）、Western blot 和 RT-PCR 用于验证 JO 的分子机制。
体内结果表明，JO（1、2、4g/kg）剂量依赖性地改善了小鼠的运动表现，降低了肺 W/D 重量比以及 IL-6 和 TNF-α的产生，但增加了 ALI 组中 IL-10 的释放。网络药理学分析表明，Toll 样受体（TLR）途径可能是 JO 治疗 ALI 的基本作用机制。免疫荧光染色和免疫共沉淀分析表明，JO 降低了 TLR4 和 MyD88 的表达水平，并减少了它们在 ALI 小鼠肺组织中的相互作用。同时，JO 减少了 NF-κB P65 的核转位和磷酸化。细胞实验结果与体内结果一致。FRET 实验也证实了 JO 干扰了 TLR4 和 MyD88 的相互作用。随后，我们还发现，JO 的六个指示性成分具有与 JO 相似的治疗效果。
综上所述，我们认为 JO 抑制了 TLR4/MyD88/NF-κB 信号通路，从而抑制了 LPS 诱导的体外和体内 ALI。阐明的机制为 JO 治疗 ALI 提供了重要的理论依据和新的治疗策略。
