Organ toxicity of Paris polyphylla in mice and the underlying hepatotoxic mechanism uncovered by an integrated network toxicology and transcriptomics.

ETHNOPHARMACOLOGICAL RELEVANCE: Paris polyphylla, a prominent traditional Chinese medicinal herb known for its heat-clearing and detoxifying properties, is widely utilized to relieve pain and treat sores, snake bites, and traumatic injuries. Recent pharmacological investigations have revealed its marked anti-tumor activity. However, as research continues to advance, its potential organ toxicity has been identified, presenting certain limitations for related studies.

PURPOSE: This study aimed to compare the toxic components, toxic doses, and organ-specific toxic responses of the cultivated and wild varieties of two P. polyphylla cultivars, and to elucidate potential mechanisms underlying variety-specific hepatotoxicity.

METHODS: Using ultra-performance liquid chromatography-tandem mass spectrometry, we identified the major active components in cultivated and wild varieties of P. polyphylla cultivars: P. polyphylla Smith var. yunnanensis (Franch.) Hand.-Mazz, referred to as PY, and P. polyphylla Smith var. chinensis (Franch.) Hara, referred to as PC. A comparative analysis was conducted to assess the differences in their contents. Additionally, we evaluated both the acute and subchronic toxicity of these varieties in mice to determine the differences in organ damage caused by the cultivated and wild varieties of PY and PC across various doses. This evaluation was based on the changes in mouse body weight, organ coefficients, and serum biochemical indicator levels, as well as histopathological alterations. Focusing on cultivated PY, we investigated its potential hepatotoxicity targets and the underlying mechanisms using a combination of network toxicology and transcriptomic techniques. The findings were validated through western blotting, terminal deoxynucleotidyl transferase dUTP nick end labeling, and transmission electron microscopy.

RESULTS: Quantitative analysis revealed that PY contained higher levels of active components than PC, with wild varieties exceeding cultivated ones. Acute toxicity test revealed that the maximum tolerated doses of the extracts from both cultivated and wild PY and PC varieties exceeded 80 g/kg body weight in mice. In the subchronic toxicity test, when the doses of PY and PC exceeded 1.95 g/kg body weight, significant increases were observed in the levels of liver and heart function biomarkers. Pathological examinations primarily revealed inflammatory changes in the liver and heart in several cases. Notably, wild varieties elicited more pronounced organ damage than cultivated ones. Comprehensive network toxicology and transcriptomic analyzes revealed that P. polyphylla intervention disrupted hepatic metabolic homeostasis through pathways that impair mitochondrial structure and function, while simultaneously activating inflammatory response pathways and promoting hepatocyte apoptosis. Enrichment analysis revealed several key targets, including STAT3. Further experimental results demonstrated that cultivated PY significantly elevated inflammatory factor levels in mice. In liver tissue, there was an aberrant activation of apoptosis and mitochondrial autophagy, accompanied by abnormal expression of related molecules, such as STAT3, Bcl-2, and BNIP3. These research findings indicate that c-PY extract stimulates abnormal mitochondrial autophagy activation, induces apoptosis and inflammatory responses in liver tissue, and ultimately leads to liver damage, potentially associated with the activation of the STAT3/BNIP3 axis. Our study also highlights the efficacy and potential of integrating network toxicology and transcriptomic analyzes in toxicological studies.

CONCLUSION: In this study, we compared the composition and toxic effects of cultivated and wild PY and PC varieties. Using transcriptomics and network toxicology, we revealed potential mechanisms of cultivated PY-induced hepatotoxicity. This research lays the foundation for determining the quality standards of P. polyphylla, evaluating the safety of its medicinal use, and elucidating the mechanisms underlying its toxicity.