Novel insights into anti-inflammatory activity-related metabolic markers and biosynthetic regulatory mechanisms between principal and lateral roots of Aconitum carmichaelii Debx. via integrated metabolomics and transcriptomics.

BACKGROUND

The principal roots (CHW) and lateral roots (FZ) of Aconitum carmichaelii Debx. are widely used medicinal herbs in Asia. Although the two roots are derived from the same plant, they exhibit differences in anti-inflammatory activity (AIA). In the market, small FZ is frequently passed off as CHW. Therefore, revealing AIA-related markers and biosynthetic regulatory mechanisms between CHW and FZ are critical for the quality control of these medicinal materials.

OBJECTIVE

To identify AIA-related markers and reveal the biosynthetic regulatory mechanisms of these markers.

METHODS

A "metabolite-gene-efficacy" correlation strategy was adopted. Firstly, verified the AIA difference between CHW and FZ via RAW264.7 cell model and Cyclooxygenase-2 inhibition assay, then integrated metabolomics and transcriptomics to screen AIA-related markers and regulatory genes, finally validated the results via qRT-PCR, AIA assay and molecular docking.

RESULTS

12 AIA-related diterpenoid alkaloid (DA) markers were identified, with 3 components as specific discriminative markers for CHW and FZ. A novel "modular regulatory" mechanism of DA markers biosynthesis was revealed: 8 DA markers in FZ were regulated by highly expressed 3‑hydroxy-3-methylglutaryl-CoA reductase in the mevalonate pathway, while 4 high-accumulation DA markers in CHW were positively driven by 1-deoxy-d-xylulose-5-phosphate reductoisomerase, 1-deoxy-d-xylulose-5-phosphate synthase, ent‑kaurene oxidase, ent‑kaurene synthase and BAHD-type acyltransferase in the methylerythritol phosphate and DA skeleton biosynthesis pathways, and 5 transcription factor families regulated the expression of the above-mentioned synthases.

CONCLUSION

This study establishes a "gene regulation → metabolite accumulation → efficacy formation" framework to explain the AIA difference between CHW and FZ, provides specific markers for their quality control, and offers novel insights for DA biosynthetic engineering.