(P23-027-23) Anthocyanin-Enriched Extracts and Pure Phenolic Compounds Reduced Intracellular Reactive Oxygen Species Induced by Particulate Matter in Dermal Fibroblasts

Objectives: Evaluate and compare the antioxidant potential of crude and anthocyanin-enriched extracts from two unique bean varieties (Phaseolus vulgaris L.) and pure phenolic compounds in HS27 and HS68 fibroblast cell lines.

Methods: Black bean (BB) and pinto bean (PB) were extracted through conventional leaching using ethanol–water (50:50 v/v). The extracts were purified through a XAD-7HP resin to obtain anthocyanin-enriched extracts (ANC). Crude extracts (SC) from both varieties were also evaluated. Delphinidin (Del), malvidin (Mal), pelargonidin (Pel), and petunidin (Pet) concentrations were measured through HPLC. Briefly, C-BB (25-400 µg/mL), C-PB (25-400 µg/mL), ANC-BB (25-400 µg/mL), and ANC-PB (25-400 µg/mL) were tested for cell viability. Intracellular ROS induced by particulate matter (PM₁₀) was determined through 2´,7´-dichlorofluorescein diacetate fluorescence assay. All extracts (100 µg/mL), ascorbic acid, gallic acid, rutin, and catechin (100 µM) antioxidant potential were evaluated. Specific markers tested the SIRT1-NF-κB pathway through western blot analyses.

Results: Del (46 mg/g SC), Mal (29.9 mg/g SC), and Pet (7.5 mg/g SC) were found in major concentrations in the ANC-BB. Pel (0.53 mg/g SC) was only identified in the ANC-PB. C-PB and ANC-PB significantly reduced cell viability to 51% in both cell lines at 400 µg/mL (p < 0.05). ANC-BB reduced it at 53% in HS27 at 400 µg/mL. ANC-BB reduced cell viability in the HS68 cell line at 47% with 400 µg/mL (p < 0.05). For intracellular ROS determination, in HS27, C-BB (40.25% ± 3.78), C-PB (32.76% ± 5.71), ANC-BB (31.18% ± 12.07), ANC-PB (19.60% ± 5.23) reduced (p < 0.05) the percentage of intracellular ROS respect to the PM₁₀ treatment (100%). In the case of HS68, C-BB (34.92% ± 4.57) and ANC-PB (28.21% ± 8.14) decreased ROS significantly (p < 0.05). SIRT1 relative expression increased to 101.4% in C-PB and 119.4% in catechin treatments.

Conclusions: Black bean and pinto bean extracts presented high concentrations of anthocyanins. In addition, the crude and anthocyanin-enriched extracts reduced the intracellular ROS production induced by PM₁₀ in dermal fibroblasts modulating the SIRT1-NF-κ-B pathway.

Funding Sources: Partially funded by CONACYT-Mexico scholarship number 901000, FORDECYT grant number 292474 and the US Department of Agriculture.