Chemicals: Folin-Ciocalteu reagent, sodium carbonate, gallic acid, trifluoroacetic and sulphuric acids, Trolox (6-hydroxy -2, 5, 7, 8- tetramethyl chromane-2-carboxylic acid), ABTS (2, 2'-azinobis (2-methylpropionamidine) dihydrochloride), 2′, 7′-Dichlorodihydrofluorescein diacetate (H2DCFDA), dexamethasone (DEX), and lipopolysaccharide (LPS) were purchased from Sigma Inc. (St. Louis, MO). Catechin, epicatechin, procyanidin B1 and B2 were purchased from Chromadex (Irvine, CA). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum, TrypLETM, TRIzol reagent, and cDNA Reverse Transcription kit were purchased from Life Technologies ( Grand Island, NY). Penicillin and streptomycin were obtained from Fisher Scientific (Atlanta, GA). All other solvents and chemicals used in this investigation were obtained from VWR International (Suwanee, GA).
Plant Material: Leaves from A. mearnsii, were obtained from Guangxi Huangmian State-owned Forest Farm (Guangxi, China). Air-dried leaves were ground into fine powders of 30–80 mesh.
Preparation of Crude Extract and Fractions: Powder leaves (30 g) was extracted twice with 500 mL of acidified 80% aqueous methanol (0.1% TFA) and then with 500 mL of 70% aqueous acetone. …show more content…
Cells were seeded in 24-well plates (5 × 105 cells/well) 24 h prior to treatment. The cells were then treated with A. mearnsii crude extract (L) and fractions (L1-L4) at predetermined doses for 1 h before elicitation with LPS at 1 µg/mL for an additional 4 h. For every experiment, one positive control (cells treated with DEX, at 10 µM) and one negative control (cells treatment with vehicle) were included. Three replicates were made for both the treatments and the controls. At the end of the treatment period, cells were harvested in TRIzol reagent for subsequent cellular RNA
Plant Material: Leaves from A. mearnsii, were obtained from Guangxi Huangmian State-owned Forest Farm (Guangxi, China). Air-dried leaves were ground into fine powders of 30–80 mesh.
Preparation of Crude Extract and Fractions: Powder leaves (30 g) was extracted twice with 500 mL of acidified 80% aqueous methanol (0.1% TFA) and then with 500 mL of 70% aqueous acetone. …show more content…
Cells were seeded in 24-well plates (5 × 105 cells/well) 24 h prior to treatment. The cells were then treated with A. mearnsii crude extract (L) and fractions (L1-L4) at predetermined doses for 1 h before elicitation with LPS at 1 µg/mL for an additional 4 h. For every experiment, one positive control (cells treated with DEX, at 10 µM) and one negative control (cells treatment with vehicle) were included. Three replicates were made for both the treatments and the controls. At the end of the treatment period, cells were harvested in TRIzol reagent for subsequent cellular RNA