Human total mmp 1

Damiana leaves powder was purchased from Ecuadorian Rainforest, LLC (Clifton, NJ, USA). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), and penicillin-streptomycin were supplied by Gibco RBL (Grand Island, NY, USA). Standard apigenin, positive control ascorbic acid, dexamethasone, and 5-diphenyltetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Human Total MMP-1 and Human Total MMP-3 ELISA kits were purchased from R&D Systems (Minneapolis, MN, USA). Organic solvents were purchased from Samchun Chemical (Seoul, Korea) and Daejung Chemical & Metal (Siheung, Korea). Inorganic salts were purchased from Sigma-Aldrich. Silica gel was purchased from Merck (Kenilworth, NJ, USA). The primary and secondary antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA), Santa Cruz Biotechnology (Santa Cruz, CA, USA), and Bio-Rad Laboratories, Inc. (Hercules, CA, USA).

NHDF (3 × 10³ cells/well) cells were seeded into 48-well plates for 24 h of incubation at 37 °C in 5% CO₂. Then, the cells were treated with extracts of skin, brain, liver, and stomach of both N. rossii and C. gunnari (NRSK, NRB, NRL, NRS, CGSK, CGB, CGL, and CGS) (25 µg/mL) in the presence or absence of UVB irradiation (125 mJ/cm²) separately, following the previously described method. After 48 h of incubation, the concentrations of MMP-1 were analyzed from the preserved supernatant using commercially available ELISA kits (Human Total MMP-1; R&D Systems Inc.) following the manufacturers’ instructions. Each sample was analyzed in triplicate.