Influence of Extraction Method on the Bioactivity of Pistacia vera L. Extracts: Metabolic and Gene Expression Changes in Colorectal Cancer Cells

Sustainable extraction methods represent a key strategy in green chemistry and nutraceutical development, aiming to replace conventional solvent-based techniques while maintaining extract quality and safety. This study compared pistachio (<i>Pistacia vera</i> L.) extracts obtained by Ultrasound-Assisted Extraction (UAE) and a classical solvent-based protocol, focusing on compositional features and biological effects. Extracts were characterized for their chemical profiles, and their impact on HCT-116 colon-derived cells was evaluated through viability assays and gene expression analysis. The UAE-derived extract, richer in carbohydrates, promoted higher cell proliferation after 72 h, whereas the classical extract upregulated <i>HMOX-1</i>, suggesting activation of antioxidant defense pathways. Moreover, UAE treatment downregulated <i>GLUT2</i> expression while modulating cytokinestranscripts, indicating a possible carbohydrate-driven immunometabolic response. Overall, these findings highlight both the advantages and limitations of green extraction approaches: while environmentally sustainable and efficient, ultrasound-assisted protocols may modify extract composition in ways that influence biological responses. Optimization of extraction parameters is therefore essential to ensure a balance between ecological sustainability, compositional integrity, and biological safety.