Helianthus annuus L. (sunflower) seed is an oilseed crop with documented anti-diabetic potential. This study employed in vitro (cell line model) to validate the anti-diabetic action of sunflower seed cultivars on the modulation of the peroxisome proliferator-activated receptor (PPAR) signalling pathway, focusing on the hub genes matrix metalloproteinase 1 (MMP1) and peroxisome proliferator-activated receptor alpha (PPARA), obtained from our previous network pharmacology study. Cytotoxicity assessment of the six cultivars investigated revealed the optimal concentrations of 100 μg/mL for AGSUN 5108 CLP and AGSUN 5270, 75 μg/mL for AGSUN 5103 CLP and AGSUN 5101 CLP, and 50 and 25 μg/mL for AGSUN 5206 CLP and AGSUN 8251, respectively. Glucose consumption was reduced in AGSUN 5103 CLP, AGSUN 8251, and AGSUN 5101 CLP treated cells compared to metformin (14.85 mmol/L) and untreated insulin-resistant HepG2 cells (17.00 mmol/L). Additionally, these cultivars upregulated the expression of MMP1 and PPARA, with AGSUN 5101 CLP exhibiting the higher expression level, showing the fold increases in MMP1 (1.88) and PPARA (4.59) expression, relative to metformin (MMP1= 8.25; PPARA = 12.38) and insulin (MMP1= 3.71; PPARA = 10.37). These findings highlight the potential of sunflower seeds, particularly cultivar AGSUN 5101 CLP, as natural therapeutic agents for diabetes through PPAR signalling pathway. Through MMP1 and PPARA modulation within PPAR pathway, the seeds may stimulate glucose uptake, insulin sensitivity, and fatty acid oxidation, which may aid in reducing the risk of type 2 diabetes mellitus (T2DM) and its associated complications. However, further pre-clinical, translational, and clinical studies are required to substantiate these findings.
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