Exploration of the pharmacological substance basis and efficacy evaluation of fufang she-she-cao (Hedyotis diffusa formula) in repressing gastric cancer.

[ETHNOPHARMACOLOGICAL RELEVANCE] Due to the low early diagnosis rate and strong heterogeneity, gastric cancer (GC), a malignant tumor of the digestive tract, still poses a serious threat to human health. Fufang She-She-Cao (Hedyotis diffusa formula, HDF), a classic ancient prescription used in Shanghai General Hospital, has been applied to prevent and treat precancerous lesions of GC for many years. However, it lacks systematic preclinical studies.

[AIM OF THE STUDY] The purpose of this study is to explore the pharmacodynamic material basis of HDF, evaluate its efficacy in GC, and elucidate its underlying pharmacological mechanisms.

[MATERIALS AND METHODS] The chemical composition fingerprint of HDF was characterized by UPLC-HRMS. Based on the chemical components identified in HDF, a content determination method was established using UPLC-MS technology. "Compounds-target-disease' network was constructed by network pharmacology analysis. The inhibitory effect of HDF on GC in vitro was detected by CCK8, colony formation assay, EdU assay, wound healing assay, transwell assay and other methods. Nude mice were used to construct a cell-derived xenograft tumor model to evaluate the anti-GC efficacy of HDF in vivo. Preliminary exploration and verification of its potential pharmacological molecular mechanism through Western blot.

[RESULTS] HDF contains 1616 chemical components, belonging to 120 categories, including Quercetin, Engelitin and Dioscin, etc. A content detection method has been successfully established for these three components, with specificity, linear relationship, precision, and stability all meeting the content determination requirements. HDF significantly inhibited the proliferation, migration, and invasion abilities of GC cells, induced apoptosis of GC cells, and also had a significant anti-tumor effect in vivo. Based on the chemical component targets obtained by network pharmacology analysis, GO and KEGG analysis of core targets showed that they were involved in multiple classic tumor-related signaling pathways and cellular biological processes. WB results showed that HDF could significantly reduce Akt protein expression, enhance GSK3β phosphorylation and degradation of β-catenin, promote DKK1 protein expression, competitively bind to Wnt protein, and collaboratively reduce the activity of Wnt/β-catenin pathway.

[CONCLUSIONS] HDF has the advantages of diverse active ingredients and multiple therapeutic targets, and it could inhibit the activity of the tumor-related classic signaling pathway Wnt/β-catenin to exert anti-GC effects. This study is the first to systematically evaluate the anti-GC effect of HDF and preliminarily explain its pharmacological mechanism, providing solid theoretical support for the clinical prevention and treatment of GC with HDF.