Identification of Anti-Inflammatory and Analgesic Mechanisms of Naru-3 Wei Pill Using Animal Models, UHPLC-QE-MS, and...

Naru-3 Wei Pill is a traditional Mongolian Medicine (TMM) that has anti-inflammatory, analgesic, and antibacterial effects. The present study aimed to demonstrate the anti-inflammatory and analgesic effect of Naru-3 Wei Pill, identify the core components and key targets and determine the pharmacological basis and mechanism of its blood-borne components.

The anti-inflammatory and analgesic effects of Naru-3 pills were studied using hot plate, tail flick, and acetic acid-induced writhing tests. The blood-borne components of Naru-3 Wei Pill were identified using ultrahigh-performance liquid chromatography with Q-Exactive mass spectrometry (UHPLC-QE-MS), and their targets were screened using ChEMBL, TCMIO, GeneCards, and TTD databases. Network construction, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, and protein-protein interaction (PPI) network was performed. Molecular dynamics methods, PCR and western blot, were used for verification of the results.

Multiple pain models results indicate that Naru-3 pills exert anti-inflammatory and analgesic effects by modulating the PI3K signaling pathways and reducing inflammation factors (interleukin [IL]-6). A total of 35 blood-borne components were identified, acting on 291 targets involving 172 pathways. The core targets included AKT1, SRC, mitogen-activated protein kinase 14 (MAPK14), and ESR1. Molecular docking and dynamics simulations conformed strong binding affinity of the complexes formed between Rhein and AKT1, MAPK1, and SRC, and between genistein and SRC. PCR and western blot confirm the regulation of AKT/PI3K pathway of Naru-3 pills in animal models.

Naru-3 pills significantly prolonged pain thresholds and reduced pain behaviors in mice. The study identified the material basis and mechanisms of its anti-inflammatory and analgesic effects, providing a foundation for further research.