An integrated RNA-Seq and network pharmacology approach for exploring the preventive effect of Corydalis bungeana Turcz. Extract and Acetylcorynoline on LPS-induced acute lung injury

Corydalis bungeana Turcz. (KDD) is a Chinese herbal medicine with anti-inflammatory, lung cleansing, detoxification and other functions. Clinically, it is commonly used to treat respiratory infections. This study uses ALI as the research model, which is consistent with the clinical use of KDD. Acetylcorynoline (AC) is the main alkaloid component of the KDD extracts, and network pharmacology studies suggest that it may be the main active ingredient in the prevention of ALI.The aim of this study is to explore the underlying mechanisms and to study the efficacy material basis of KDD in anti-ALI effect by LPS-induced mice and using a combination of RNA sequencing (RNA-Seq) technology and network pharmacology.Establish a mouse model of ALI by intraperitoneal injection of LPS (5 mg/kg). The main active ingredients of KDD were identified and analyzed by high performance liquid chromatography with quadrupole time-of-flight mass spectrometry (HPLC-QTOF-MS) and network pharmacology. IL-18, IL-1β, and IL-6 levels in serum and bronchoalveolar lavage fluid (BALF), lung histopathological changes, and lung myeloperoxidase (MPO) activity were assessed. We investigated the possible molecular mechanisms of KDD and AC in an LPS-induced mouse ALI models with RNA-Seq technology. In addition, the anti-inflammatory effect of AC was verified in vitro by establishing an LPS-stimulated RAW264.7 inflammation model. Molecular docking further validated AC as the efficacy material basis of KDD in anti-ALI.Based on HPLC-QTOF-MS technology and network pharmacology, KDD is more strongly associated with lung tissue, and that AC may be the main active ingredient of KDD. Subsequently, in vivo experiments results showed that KDD and AC reduced the levels of pro-inflammatory cytokines in serum and BALF, reduced MPO levels and reduced inflammatory damage in the lungs. To elucidate its underlying mechanism, based on RNA-Seq analysis techniques performed in lung tissue, enrichment analysis showed that KDD and AC intervened through the NLR signaling pathway, thereby mitigating LPS-induced ALI. Then, RT-qPCR, IF, WB and other technologies were used to verify the anti-ALI core difference genes of KDD and AC from the gene transcription and protein expression levels of the NLR signaling pathway, and confirmed the anti-ALI. In vitro experimental results also showed that AC has anti-inflammatory effects in RAW264.7. Finally, the biotransformation and molecular docking results also further indicated that AC is the active ingredient of KDD in anti-ALI.Studies have shown that KDD has a good therapeutic effect on ALI, and AC is the main pharmacodynamic material basis for its therapeutic effect in ALI.