Therapeutic and toxicity targets for Euphorbia Kansui L. in treating hepatocellular carcinoma ascites revealed by ingredients pattern analysis accompanying vinegar processing.

Euphorbia Kansui L. is effective in managing ascites resulting from hepatocirrhosis and hepatoma, however, it has some toxicity. Vinegar processing, a traditional detoxification method, can effectively reduce the toxicity of raw Kansui. However, the underlying mechanisms of Kansui's therapeutic and toxicological effects remain unclear. This topic was investigated through an integrated approach in current study. Firstly, the extract of Kansui was proved to have an inhibitory effect on the proliferation of various tumor cells, particularly liver cancer cells. Then HPLC-based ingredients pattern analysis on raw and processed Kansui indicated that there was a significant reduction for triterpenoids and jatrophane-type diterpenoids during the processing, in contrast, a mild reduction for ingenane-type diterpenoids was observed during the same processing. From network pharmacology analysis inferred that terpenoids from Kansui are involved in tumor signaling pathways and immune activation pathways and so on, may contribute to therapeutic effect of alleviate malignant ascites. Inflammatory response pathway and cellular response to chemical stimulus may contribute to the herb's toxicity such as gastrointestinal irritation. The core targets in the protein-protein interaction network were then validated by molecular docking, SPR experiments and enzymatic experiments. Enzyme inhibition assays confirming that extract of vinegar-processed Kansui exhibits reduced inhibitory effects on cholinesterases (BCHE and ACHE), aligning with predicted detoxification mechanisms. Besides, SPR analysis revealing weaker binding affinity of processed Kansui to the toxicological target ACHE, supporting the detoxification mechanism at the molecular level. These findings are meaningful for understanding the mechanism for Kansui's therapeutic and toxicological effects, meanwhile to ensure the safety of Kansui in medication.