Chemotype-Resolved Cytoprotection of CAM Hydrothermal Extracts Validated in 2D/3D Fibroblast Models and Supported by...

Area of Science:

• Biochemistry
• Plant Science
• Pharmacology

Background:

• Crassulacean Acid Metabolism (CAM) plants exhibit unique stress adaptations.
• The relationship between CAM plant chemotypes and their bioactivity is not well understood.
• Investigating CAM plant extracts for therapeutic potential is warranted.

Purpose of the Study:

• To explore the chemotype-bioactivity relationships of hydrothermal extracts from four CAM plant species.
• To identify specific chemotypes responsible for antioxidant properties.
• To validate antioxidant effects using in vitro and in silico methods.

Main Methods:

• Gas Chromatography-Mass Spectrometry (GC-MS) metabolomics for chemotype analysis.
• Principal Component Analysis (PCA) and marker analysis to classify extracts.
• 2D cell culture (NIH/3T3 fibroblasts) and 3D bioprinted tissue models for functional validation.
• In silico molecular docking to predict bioactive compounds and targets.

Main Results:

• Three distinct chemotypes were identified: acid-rich, sugar-rich, and lipid/sterol-enriched.
• Acid-rich extracts (from Bulbophyllum drymoglossum and Crassula rupestris) significantly reduced oxidative stress (ROS) and preserved mitochondrial membrane potential in fibroblasts.
• Antioxidant effects were confirmed in a 3D bioprinted tissue model, demonstrating high biocompatibility.
• In silico analysis identified phaselic acid as a potential bioactive marker interacting with Keap1, COX-2, and MMP-1.

Conclusions:

• CAM plant extracts can be categorized into distinct chemotypes with varying bioactivities.
• Acid-rich CAM plant extracts possess significant antioxidant and cytoprotective properties.
• Phaselic acid is a promising candidate for the antioxidant activity observed in specific CAM plant extracts.
• This study provides a framework for leveraging CAM plant metabolites in antioxidant strategies.