Chlorogenic acid

Caffeine (C0750 Sigma Aldrich, Saint Louis, MO, USA) and chlorogenic acid (C3878 Sigma Aldrich) were dissolved in deionized water to prepare stock solutions and then sterilized by filtration using nitrocellulose membrane (0.25 µm) (Figure S1). Glutathione monoethyl ester (353905 Sigma Aldrich) and N-acetyl-L-cysteine (A7250 Sigma Aldrich) were used as established antioxidant compounds (Figure S1) and dissolved according to the manufacturer’s instructions. Menadione sodium bisulfite (M2518 Sigma Aldrich) and diclofenac sodium salt (D6899 Sigma Aldrich) were used as positive controls for ROS production. Palmitate stock solution (20 mmol/L) was prepared using sodium palmitate (Sigma Aldrich P9767) and sodium oleate in an aqueous solution of bovine serum albumin (BSA fatty acid-free) as previously described [27 (link)]. Free fatty acid mix (FFA) was prepared by mixing oleate and palmitate solutions at a 2:1 ratio, obtaining a final concentration of 0.75 mmol/L (0.5 mmol/L: oleate and 0.25 mmol/L: palmitate). Saturated 2-SG (2-Stearolylglycerol 36484), monounsaturated 2-OG (2-Oleoylglycerol M2787, Sigma Aldrich) and Stearoyl-CoA Desaturase 1 inhibitor (SCD-1 inhibitor 569406, Sigma Aldrich) were dissolved according to the manufacturer’s instructions.

The ethanol of agricultural origin used for extraction was of analytical grade and was obtained from the MV Group (Kaunas, Lithuania). The reagents used in this study were as follows: acetonitrile (HPLC grade) Sigma-Aldrich Company (Poznan, Poland), acetone (HPLC grade) Sigma-Aldrich Company (Poznan, Poland), deionised water, ethyl acetate (HPLC grade) Merck (Poznan, Poland), polyphenols standards (epigallocatechin, catechin, chlorogenic acid, epigallocatechin gallate, p-coumaric, quercetin-3-O-rutinoside, kaempferol-3-O-glucoside, myricetin, quercetin, kaempferol, quercetin-3-O-glucoside, cyanidin-3-O-glucoside, cyanidin-3-O-rutinoside, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)Sigma-Aldrich Company (Poznan, Poland), sulfuric acid 98% Merck Company (Poznan, Poland), DMEM medium GlutaMAX™ Supplement, fetal bovine serum (FBS), penicillin, and streptomycin (Gibco, TermoFisher Scientific, Waltham, MA, USA).

All reagents and solvents were of analytical grade unless otherwise specified and used without further purification. 2,2-Diphenyl-1-picryhydrazyl radical (DPPH) was purchased from Alfa Aesar (London, UK), ethanol 96% and ethanol absolute from Carlo Erba (Rodano, MI, Italy). Gallic acid (3,4,5-trihydroxybenzoic acid), quinolinic acid (pyridine-2,3-dicarboxylic acid), ferrous sulfate heptahydrate, hydrogen peroxide (30% w/w), ethanol, ethyl acetate, ethyl lactate, sodium carbonate and Folin-Ciocalteu reagent were purchased from Sigma–Aldrich (Milan, Italy). DMPO (5,5-dimethyl-1-pyrroline N-oxide) was purchased from Enzo Life and used without further purification. Water was purified with a Milli-Q system from Millipore (Millipore Corporation, Billerica, MA, USA).
For chromatographic analyses, HPLC grade water, acetonitrile and methanol were purchased from VWR (Milan, Italy). High purity commercial standards luteolin, luteolin-7-O-glucoside and apigenin-7-O-glucoside were provided by Extrasynthese (Lyon, France), whilst chlorogenic acid, hydroxytyrosol, oleuropein and 3,4-dihydroxyphenilacetic acid (DOPAC) were provided by Sigma-Aldrich (Milan, Italy).
BPOV (bis(maltolate)oxidovanadium(IV)) and BMOV (bis(picolinate)oxidovanadium(IV)) were synthesized according to the methods reported in the literature (Orvig et al. 1995 (link); Lodyga-Chruscinska et al. 2011 (link)).
Human colon fibroblast cell line CCD-33Co (CRL-1539™) and preadipocyte cell line isolated from adipose fat tissue hTERT A41hBAT-SVF (CRL-3385™) were purchased from American Type Culture Collection (LGC Standards, Milan, Italy).
DMEM high glucose, fetal bovine serum (FBS), RPMI 1640, Penicillin/Streptomycin, L-glutamine, and 25% Trypsin–EDTA were purchased from Euroclone (Pero, MI, Italy).
Human insulin solution (I9278), dexamethasone (D4902), 3-isobutyl-1-methylxanthine (IBMX, I5879), 3,3',5-triiodo-L-thyronine sodium salt (T3, I6397), indomethacin (I7378), D-pantothenic acid (P5155), biotin (B4639), D-( +)-glucose (G7021), Oil Red O solution (O1391), Formalin solution neutral buffered 10% (HT5012), Bovine Serum Albumin (A-3059), and hematoxylin solution (MHS16) were Sigma-Aldrich products (Milan, Italy).
Rabbit pAb to Glucose Transporter GLUT4 (ab216661), Rabbit specific HRP/AEC (ABC) Detection IHC Kit (ab64260), Aqueous Mounting Medium (ab128982) were Abcam Products (Prodotti Gianni, Milan, Italy).

Chemicals were purchased as follows: cannabidiol (2) from Knowde (USA), quercetin (9) from Koch-Light laboratories Ltd. (Suffolk, UK), moracin T (12) and mullberofuran K (16) from ChemFaces (Wuhan, China), doxycycline (17) from Fagron (Olomouc, Czech Republic), caffeine (18) from Lancaster (Morecambe, UK), chlorogenic acid (19), and epigallocatechin gallate (20) from Sigma-Aldrich (St. Louis, MO, USA).
Mueller–Hinton medium and microbial agar were obtained from Sigma-Aldrich. QS AI-2 Bioassay (AB) medium was composed of NaCl (Penta, Praha, Czech Republic), MgSO₄·7H₂O (Lach-Ner, Neratovice, Czech Republic), casamino acids (Gibco-Life technologies corporation—Thermo Fisher, Waltham, MA, USA), KOH (Sigma-Aldrich, St. Louis, MO, USA), L-arginine and thiamine (VWR Chemicals, Radnor, PA, USA), riboflavin (Sigma-Aldrich, St. Louis, MO, USA), KH₂PO₄ (Penta, Praha, Czech Republic), K₂HPO₄ (Penta, Praha, Czech Republic), and glycerol (Sigma-Aldrich, St. Louis, MO, USA).
Preparation of AB (AI-2 bioassay) medium: 300 mM NaCl, 0.5 mM MgSO₄, and 2mg/mL of casamino acids were mixed with distilled water. KOH was added to adjust the pH to 7.5. The mixture was then autoclaved for 30 min. A total of 0.5 mM L-arginine, 20 μg/mL thiamine, 2 μg/mL riboflavin, 2.5 mM K₂HPO₄, 2.5 mM KH₂PO₄ and 0.5% glycerol were each dissolved separately, in distilled water. These solutions were filtered through a 0.2 μm microfilter syringe and subsequently added to the rest of the AB medium for V. harveyi MM30 [14 (link)].

Folin–Ciocalteu’s phenol reagent, Ethyl acetate, n-hexane, and methanol were purchased from Merck (Darmstadt, Germany). 1,1-diphenyl-2-picrylhydrazyl (DPPH), potassium persulfate (K₂S₂O₈), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonate) (ABTS), 1,3,5-triphenyltetrazolium chloride (TPTZ), Trolox (6-hydroxy2,5,7,8-tetramethylchroman-2-carboxylic acid), FeSO₄·7H₂O, gallic acid, cinnamic acid, syringic acid, ferulic acid, chlorogenic acid, caffeic acid, p-coumaric acid, benzoic acid, pinocembrin, rutin, quercetin, luteolin, kaempferol, epicatechin, catechin, apigenin, myricetin, isorhamnetin, taxifolin, chrysin, galanganin, genistein, and hesperetin were all purchased from Sigma Chemical Co. (St. Louis, MO, USA).

Gallic acid (CAS Number 149-91-7), chlorogenic acid (CAS Number 327-97-9), oenothein B (CAS Number 104987-36-2), myricitrin (CAS Number 17912-87-7), quercetin-3-O-glucuronide (CAS Number 22688-79-5), hyperoside (CAS Number 482-36-0), ellagic acid (CAS Number 476-66-4), kaempferol-3-O-glucoside (CAS Number 480-10-4), quercitrin (CAS Number 522-12-3), myricetin (CAS Number 529-44-2), kaempferol-3-O-rhamnoside (CAS Number 482-39-3), quercetin (CAS Number 117-39-5), kaempferol (CAS Number 520-18-3), and oenothein B were purchased from Merck (Milan, Italy). myricetin-3-O-galactoside (CAS Number 15648-86-9) and kaempferol-3-O-glucuronide (CAS Number 22688-78-4) were purchased from Extrasynthese (Genay, France). Individual stock solutions of each analyte, at a concentration of 1000 µg/mL, were prepared by dissolving pure standard compounds in HPLC-grade methanol:water (80:20) and storing them in glass-stoppered bottles at 4 °C. Regarding ellagic acid, the stock solution was prepared by dissolving 5 mg in 25 mL of an HPLC-grade acetone:acetonitrile:ethanol mixture (1:1:1). HPLC-grade methanol was purchased by Carlo Erba (Milan, Italy), while HPLC-grade formic acid (99%) was acquired from Merck (Darmstadt, Germany), as well as the resin Amberlite XAD7HP. A Milli-Q SP Reagent Water System (Millipore, Bedford, MA, USA) was employed to produce ultrapure water. All the solutions were filtered with Phenex™ RC 4 mm 0.2 µM syringeless filters (Phenomenex, Castel Maggiore, Italy) prior to the analysis.