Prediction of bioactive compounds hepatotoxicity using in silico and in vitro analysis

• Published in: Applied biological chemistry 2024, 67(0), , pp.1-13
• Main Authors: Choi, Kwanyong, Lee, Soyeon, Yoo, Sunyong, Han, Hyoung-Yun, Park, Soo-yeon, Kim, Ji Yeon
• Format: Article
• Language: English
• Subjects: Apoptosis; Applied Microbiology; Assaying; Bioactive compounds; Biological activity; Biological Techniques; Bioorganic Chemistry; Chemistry; Chemistry and Materials Science; Drug-induced liver injury; Hepatotoxicity; HepG2; Herbs; In silico; Injury analysis; Morphology; Piperine; Prediction; Prediction models; Side effects; Transcriptomics; Viability; 농학
• ISSN: 2468-0834; 2468-0842
• DOI: 10.1186/s13765-024-00961-z

The leading safety issue and side effect associated with natural herb products is drug-induced liver injury (DILI) caused by bioactive compounds derived from the herb products. Herein, in silico and in vitro analyses were compared to determine the hepatotoxicity of compounds. The results of in silico analyses, which included an integrated database and an interpretable DILI prediction model, identified calycosin, biochanin_A, xanthatin, piperine, and atractyloside as potential hepatotoxic compounds and tenuifolin as a non-hepatotoxic compound. To evaluate the viability of HepG2 cells exposed to the selected compounds, we determined the IC 50 and IC 20 values of viability using MTT assays. For in-depth screening, we performed hematoxylin and eosin-stained morphological screens, JC-1 mitochondrial assays, and mRNA microarrays. The results indicated that calycosin, biochanin_A, xanthatin, piperine, and atractyloside were potential hepatotoxicants that caused decreased viability and an apoptotic phase in morphology, while these effects were not observed for tenuifolin, a non-hepatotoxicant. In the JC-1 assay, apoptosis was induced by all the predicted hepatotoxicants except atractyloside. According to transcriptomic analysis, all the compounds predicted to induce DILI showed hepatotoxic effects. These results highlighted the importance of using in vitro assays to validate predictive in silico models and determine the potential of bioactive compounds to induce hepatotoxicity in humans.