Exploring the mechanisms of Yang Wei Shu granule for the treatment of chronic atrophic gastritis using UPLC-QTOF-MS/MS, network pharmacology, and cell experimentation.

Ethnopharmacological Relevance: Chronic atrophic gastritis (CAG) is a global disease of the digestive system and is an important precancerous lesion in the development of gastric cancer. Yang Wei Shu granule (YWSG), which evolved from the formula 'Warm Stomach Soup' of the Jin and Yuan Dynasties in China, is frequently used as a classic herbal compound in the treatment of CAG. However, the active ingredients and mechanisms by which it works are not clear.

Aim Of The Study: To elucidate the chemical composition of YWSG and investigate the potential mechanisms of YWSG on CAG by composition analysis, network pharmacology and cellular experimental studies.

Materials And Methods: The chemical and blood-entry constituents of YWSG were analyzed by ultra-high performance liquid chromatography-Quadrupole tandem time-of-flight mass spectrometry (UPLC-QTOF-MS/MS). Subsequently, potential targets of YWSG for CAG treatment were identified through utilization of publicly available online resources. The YWSG-component-target-pathway network and protein-protein interaction (PPI) network were constructed using Cytoscape software. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of potential targets was performed using the DAVID database. Finally, a cellular model of lipopolysaccharide (LPS)-activated RAW 264.7 macrophages was established and validated by in vitro experiments.

Results: A total of 150 compounds in YWSG and 47 blood-entry constituents were identified by using UPLC-QTOF-MS/MS. Based on network pharmacology, a total of 132 target genes were identified as being involved in the therapeutic effect of YWSG on CAG. Network pharmacology and molecular docking results suggest that AKT1, PIK3CA, PTPN11, SRC and STAT3 may be potential targets of YWSG for the treatment of CAG. Cellular experiments showed that the YWSG-containing serum had no cytotoxic effect on RAW264.7 cells and could inhibit nitric oxide (NO) production and the expression of pro-inflammatory factors TNF-α, IL-6, and IL-1β. Additionally, it was observed to promote the expression of the anti-inflammatory factor IL-10 in LPS-stimulated RAW264.7 cells. The immunofluorescence results showed that YWSG treated CAG by inhibiting the PI3K-Akt pathway.

Conclusions: The mechanism of action of YWSG on CAG was preliminarily elucidated using UPLC-Q-TOF-MS/MS, network pharmacology and in vitro experiments. The main active components and potential targets of YWSG were investigated, providing a scientific basis for further research.