Sodium glycocholate

Human primary RPE cells (HRPEpiC) were purchased from ScienCell (Carlsbad, CA, USA) and maintained in epithelial cell medium (ScienCell). Rhesus macaque choroidal endothelial cells (RF/6A) were purchased from American Type Culture Collection (Manassas, VA, USA). RF/6A cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM, Gibco, Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, Invitrogen), 100 units/mL penicillin, and 100 µg/mL streptomycin (Gibco, Invitrogen,). RPE cells were cultured for two weeks prior to experiments, and experiments were conducted in epithelial cell medium. CECs were serum-starved in DMEM with 2% FBS overnight prior to experiments, and all RF/6A experiments were performed in DMEM with 0.5% FBS. Cells were incubated at 37 °C, 5% CO₂, 20.9% O₂, and 95% relative humidity. Sodium glycocholate (Sigma-Aldrich #G7132, St. Louis, MO, USA), sodium glycodeoxycholate (Calbiochem, Millipore #361311, Burlington, MA, USA), and sodium glycoursodeoxycholate (Toronto Research Chemicals #G679545, Toronto, ON, Canada) were dissolved in water to make 10 mM stock solutions for use in experimental assays.

The fruiting body of D. indusiata was bought from the local supermarket in Ya’an, China. The sample was dried, grounded, and sieved through a 60-mesh screen. The powders were stored at −20 °C before use. Carboxymethyl cellulose (CM-cellulose), cholesterol, sodium deoxycholate, sodium taurocholate, sodium glycocholate, sodium cholate, oleic acid, 1-phenyl-3-methyl-5-pyrazolone (PMP), monosaccharide standards, Griess reagent, 2,2-diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl (DPPH), and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) were obtained from Sigma-Aldrich (Sigma, St. Louis, MO, USA). A free cholesterol assay kit was obtained from Solarbio (Solarbio, Beijing, China).

Sodium deoxycholate (NaDC; 97%), sodium taurodeoxycholate (NaTDC; 95%), sodium taurocholate (NaTC; ≥97%), sodium cholate (NaC; 97%), sodium glycocholate (GC; ≥97%), sodium glycodeoxycholate (GDC; ≥97%) and l‐α‐phosphatidylcholine from dried egg yolk (≥50%) were used as purchased from Sigma Aldrich, Dorset, UK for the preparation of stock solutions of mobile phase. Analytical grade 4‐(2‐hydroxyethyl)‐1‐piperazineethanesulfonic acid (HEPES buffer) was purchased from Sigma Aldrich, Dorset, UK. The compounds considered in this work were caffeine 97% (Sigma Aldrich, Dorset, UK), fenoprofen 97% (Fluka, Dorset, UK), acetaminophen 99% (Sigma Aldrich, Dorset, UK), ketoprofen 98% (Sigma Aldrich, Dorset, UK), phenylbutazone 99% (Sigma Aldrich, Dorset, UK), fluconazole 98% (Sigma Aldrich, Dorset, UK), carbamazepine 99% (Sigma Aldrich, Dorset, UK), cimetidine (Sigma Aldrich, Dorset, UK), naproxen 98% (Sigma Aldrich, Dorset, UK), terbutaline 96% (Sigma Aldrich, Dorset, UK), zolmitriptan >98% (Sigma Aldrich, Dorset, UK), salicylic acid 99% (Fisher Scientific, Loughborough, UK), ibuprofen 98% (BASF, Cheshire, UK), acetyl salicylic acid 99% (Acros Organics, Geel, Belgium), diclofenac 98% (TCI Europe, Zwijndrecht, Belgium), flurbiprofen 98% (TCI Europe), nicotinic acid >98% (Sigma Aldrich, Dorset, UK) and theophylline 98%, (TCI, Oxford, UK).