Study on the pharmacodynamic material basis of Buthus martensii Karsch in ameliorating chronic atrophic gastritis: Screening and experimental validation of protein-peptide bioactive components based on 4D label-free proteomics.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE: Chronic atrophic gastritis (CAG) is a digestive system disorder characterized by persistent inflammation and damage to the gastric mucosa, marking a critical stage in the inflammation-carcinoma transition. Buthus martensii Karsch (BmK) has been clinically used to alleviate CAG and impede the progression to carcinoma. However, the active components and pharmacological mechanisms of BmK remain largely unexplored. AIM OF THE STUDY: This study integrated proteomic analysis with experimental validation to provide an initial characterization of BmK's proteomic profile, identify its active components, and elucidate the therapeutic pathways involved in its action against CAG. MATERIALS AND METHODS: In vivo experiments were conducted to assess the therapeutic efficacy and safety of BmK in treating CAG. High-throughput sequencing was subsequently employed to construct BmK's transcriptome and develop a translatable protein database. To identify protein-peptide components, 4D label-free mass spectrometry-based quantitative proteomics was utilized. Potential functional peptide segments were screened, and their therapeutic effects and mechanisms were evaluated through in vivo experiments. RESULTS: In vivo efficacy tests revealed that the bioactive components of BmK significantly enhanced gastric function and reduced inflammation in CAG-afflicted mice. Safety assessments indicated that, at the standardized dosage, BmK did not impair liver function, kidney function, or motor behavior. Transcriptomic and proteomic analyses identified 17,604 genes, 12,368 proteins, and 33,834 peptides in BmK. Differential proteomic analysis showed distinct protein profiles across different medicinal parts and dosage forms of BmK, with the scorpion tail powder identified as the main bioactive component. The peptide TRINITY_DN1656_c0_g1.p1 was highlighted as a key bioactive protein-peptide. In vitro synthesis of peptide STP1656 based on TRINITY_DN1656_c0_g1.p1 demonstrated its ability to improve pathological damage and inflammation in CAG mice. Further experiments indicated that STP1656 exerted therapeutic effects by modulating glycolysis-related proteins (PKM2, HK2) and apoptosis-related proteins (Bcl-2, Bax). CONCLUSION: This study provides novel insights into the material basis and molecular mechanisms through which BmK ameliorates CAG, primarily by regulating the glycolytic pathway and apoptosis. Furthermore, it highlights the potential of bioactive protein-peptides as a promising therapeutic approach.