Application of Chemometric Analysis With Acute Toxicity Tests of Zebrafish and Network Pharmacology for Comparison of Different Species: Rhubarb as an Example.

Abstract

This study systematically evaluated the quality variations of Rheum spp. from different botanical origins through an integrated approach combining chemical characterization and biological validation. Chemometric analysis of chromatographic profiles effectively categorized the samples into two distinct groups, successfully differentiating Rheum tanguticum Maxim. ex Balf. from Rheum officinale Baill., and Rheum palmatum L., though the latter two species showed overlapping chemical characteristics. In the zebrafish acute toxicity test, the half-death rate of R. tanguticum was much lower than that of R. officinale and R. palmatum, further indicating that the classification was reasonable and reliable. Based on chemometrics and toxicity test results of zebrafish, four components of sennoside B, sennoside A, aloe-emodin 8-O-glucoside, and chrysophanol were selected as potential toxicity markers of rhubarb. Our findings suggest that rhubarb-induced hepatotoxicity likely results from multicomponent/multi-target synergistic interactions, particularly through modulation of PI3K-AKT/mTOR signaling, matrix remodeling, and stress response modulation. Molecular docking confirmed differential binding affinities, with chrysophanol and aloe-emodin 8-O-glucoside demonstrating strong target engagement. This integrated approach not only establishes a reliable quality assessment protocol for rhubarb species differentiation but also provides mechanistic insights into the complex toxicological profile of these medicinal plants, advancing our understanding of their pharmacotoxicological characteristics.