(P04-053-24) Trigonelline Prevents High-Glucose-Induced Endothelial-to-Mesenchymal Transition by Suppression of Oxidative Stress and Preservation of ATP and Mitochondrial Function

Objectives: Trigonelline is a bioactive alkaloid found in a variety of plants. Recent evidence has shown its anti-inflammatory, anti-oxidative and anti-fibrotic activities in several disease models. The potential therapeutic benefits of trigonelline in diabetes have also been documented but with unclear molecular mechanisms. This study investigated the molecular mechanisms underlying the protective effects of trigonelline on high glucose (HG)-induced endothelial cell deteriorations using a proteomics approach followed by functional validations.

Methods: Human endothelial (EA.hy926) cells were treated with 100 µM trigonelline for 24 h and subjected to label-free quantitative proteomics using nanoLC-ESI-Qq-TOF MS/MS compared with the untreated cells. Functional enrichment analyses of the significantly altered proteins were done by using the DAVID database (https://david.ncifcrf.gov/) and the REVIGO tool (https://revigo.irb.hr/). Functional validations were then performed to confirm these trigonelline-induced biological processes and molecular functions.

Results: Label-free quantitative proteomics revealed 69 significantly altered (40 downregulated and 29 upregulated) proteins in the trigonelline-treated cells. Functional enrichment analyses suggested the involvement of these altered proteins in several biological processes and molecular functions, including cell-cell adhesion, cell cadherin binding, redox homeostasis, ATP metabolic process and ATP hydrolysis activity. Quantitative analysis of the cell spindle index showed that HG caused cell elongation. In concordance, an immunofluorescence study demonstrated the HG-induced decreased levels of endothelial markers (PECAM-1 and VE-cadherin) and increased levels of mesenchymal markers (fibronectin and vimentin). Oxyblot analysis revealed the HG-induced increase of oxidized proteins. By contrast, intracellular ATP and mitochondrial membrane potential were reduced by HG. However, all of these HG-induced detrimental effects were successfully prevented by trigonelline.

Conclusions: These data have demonstrated that trigonelline prevents HG-induced EndMT via suppression of oxidative stress and preservation of intracellular ATP and mitochondrial function.

Funding Sources: None