Regulatory effect of yerba mate (Ilex paraguariensis) extracts on proliferation and metabolic activity on human epithelial renal non-tumor and tumor cell lines

FERRANDO, MATÍAS; SANTIANO, FLAVIA; CANO, ROCÍO; SANHUEZA, MARÍA DE LOS ÁNGELES; FERNANDEZ, MARÍA DE LOS ÁNGELES; ESPINO, MAGDALENA; CARÓN, RUBÉN WALTER; PISTONE-CREYDT, VIRGINIA; LÓPEZ-FONTANA, CONSTANZA MATILDE

San Juan

Congreso; XLI Reunión Anual de la Sociedad de Biología de Cuyo; 2023

Sociedad de Biología de Cuyo

Yerba mate (I. paraguariensis), a native species from South America, has demonstrated satisfactory effects in the prevention and treatment of health disorders. A recent study by Fernández et al. 2023 reports an anti-proliferative in vitro effect on prostate tumor cells of yerba mate extracts in an aqueous phase and in Natural Deep Eutectic Solvents (NADES: 0,10 g citric acid, 0,37 g glycerol and 0,53 g water). In this work, we evaluated the effect on proliferation and metabolic activity of these extracts on human renal epithelial non-tumor (HK-2) and tumor (786-O) cell lines. Yerba mate aqueous extract was solubilized in DMEM F12 or RPMI cell culture medium with 1% of FBS at 15 mg/mL, from this solution, an increasing dilution curve was formulated (starting at 3.12 to 0.095 mg/mL). The optimal yerba mate-CGH extract was solubilized in DMEM F12 or RPMI cell culture medium at 12.5 mg/mL, from this solution increasing dilution curve was formulated (starting at 2.6 to 0.079 mg/mL). We treated HK-2 and 786-O cells with various dilutions of yerba mate extracts, and subsequently, we assessed proliferation using MTT, evaluated viability by Trypan blue staining, and conducted Western Blot analysis to assess the protein expression of: 1) apoptotic proteins: cleaved and total PARP and Caspase 3, 2) cell cycle regulatory proteins: cyclin D1, retinoblastoma protein, p21, and 3) PCNA. Statistical differences were assessed by one-way ANOVA. The yerba mate extracts in the aqueous phase exhibited an anti-proliferative impact on HK-2 cells at a concentration of 1.56 μg/mL, leading to a significant reduction in their viability. Interestingly, when these cells were exposed to yerba mate extracts in combination with NADES, we observed differences in viability and metabolic activity even at lower concentrations (0.325 mg/mL) compared to the control group. As for NADES itself, no significant differences were observed between treatments. Regarding the 768-O cell line, yerba mate in the aqueous phase did not show an anti-proliferative effect. However, Yerba mate extracts dissolved in NADES exerted an anti-proliferative effect at 0.325 mg/mL, leading to a significant reduction in metabolic activity compared to control. NADES alone decreased the proliferation and viability of 786-O cells at the highest concentration (2.6 mg/mL). We observed a significant increase in the expression of p21 proteins (p