Illuminating the black box of traditional Chinese medicine: The pivotal role of transgenic fluorescent zebrafish.

ETHNOPHARMACOLOGICAL RELEVANCE: Traditional Chinese Medicine (TCM) holds significant therapeutic potential, yet its modernization is hindered by the inherent complexity of its formulations and uncertainties surrounding the precise mechanisms of action, efficacy, and safety profiles of its components. The zebrafish (Danio rerio) model, particularly transgenic fluorescent zebrafish (TFZ), has emerged as a powerful tool to address these challenges, offering a vertebrate model that combines genetic tractability with real-time in vivo visualization capabilities. AIM OF THE STUDY: This review aims to highlight the role of TFZ in advancing TCM research, particularly in drug screening, efficacy evaluation, and toxicity assessment. We systematically catalog the key transgenic lines used to assess the therapeutic potential of TCM and natural products (NPs) in various disease areas. METHODS: A comprehensive literature search was conducted across multiple databases (PubMed, Web of Science, Scopus, CNKI) from January 2000 to April 2025, using core keywords related to transgenic zebrafish, fluorescence imaging, TCM, NPs, and specific biological functions. Studies utilizing TFZ models to evaluate TCM efficacy and toxicity were included, while non-zebrafish models and studies lacking mechanistic insights were excluded. Key data were systematically extracted and categorized by disease area, with a focus on mechanistic insights. Visualization of experimental workflows and schematics was created using BioRender.com. RESULTS: This review underscores the pivotal role of TFZ in advancing TCM research and systematically organizes the key transgenic lines used to evaluate TCM and NPs across multiple domains, including anti-angiogenesis, inflammation suppression, cardiovascular and cerebrovascular protection, hepatoprotection, and neuroprotection. Specific transgenic lines, such as Tg(fli1a:EGFP) for angiogenesis studies and Tg(lyz:EGFP) for inflammation studies, have been instrumental in evaluating the therapeutic effects of various TCM compounds and extracts. Artificial intelligence (AI)-driven tools have further enhanced the efficiency and accuracy of these studies by automating data analysis and reducing subjectivity. CONCLUSION: The integration of TFZ models with AI technology has significantly advanced TCM research, providing a robust platform for high-throughput screening, efficacy evaluation, and toxicity assessment. This approach enables real-time mechanistic insights, deepening​ the understanding of TCM therapeutics.