[Investigation on anti-gastric cancer mechanism of combined prescription of modified Liangfu and Jianpi Yiqi based on zebrafish model and proteomics technology].

This study aims to explore the anti-gastric cancer mechanism of combined prescription of modified Liangfu and Jianpi Yiqi(CPMLJY) using a zebrafish xenograft model with gastric cancer, ultra-performance liquid chromatography(UPLC), and data-independent acquisition(DIA) proteomics. The active ingredients of CPMLJY were identified by UPLC. A zebrafish xenograft model was established by microinjecting human gastric cancer HGC-27 cells into the yolk sac of wild-type AB strain zebrafish. The maximum tolerated concentration(MTC) of CPMLJY was measured by a gradient dilution method. Zebrafish were randomly divided into normal control group, model control group, capecitabine group, and low-(1/4 MTC), medium-(1/2 MTC), and high-dose(MTC) groups of CPMLJY, with 30 zebrafish in each group. After 48 h at 35 ℃, the effect of CPMLJY on tumor proliferation and apoptosis was assessed via fluorescence labeling. Transgenic zebrafish(T-cell-red and vascular-green Fil-1 strains) were used for modelling to evaluate immune response and angiogenesis. DIA proteomics was used to reveal potential anti-gastric cancer mechanisms. The results showed that the main active ingredients of CPMLJY were flavonoids, steroidal saponins, and triterpenoid saponins. The medium-and high-dose of CPMLJY significantly inhibited tumor growth(P<0.05). High-dose of CPMLJY significantly increased the number of T cells, while medium-dose of CPMLJY significantly suppressed angiogenesis and induced apoptosis(P<0.05). Proteomics identified 143 upregulated and 233 downregulated proteins(P<0.05). CPMLJY exerted anti-tumor effects by modulating adipocyte lipolysis, forkhead box O(FoxO), wingless-related integration site(Wnt), and peroxisome proliferator-activated receptor(PPAR) signaling pathways, and amino acid metabolism(P<0.05). These findings support further clinical translation of CPMLJY for gastric cancer therapy.