Movas

Human aorta endothelial cells line (HAEC) was obtained from Lonza (Basel, Switzerland). Vascular smooth muscle cell line (MOVAS) was purchased from American Type Culture Collection (ATCC, Rockville, Maryland, MD, USA). HAECs were maintained in supplemented endothelial growth medium EGM-2 (Lonza, Basel, Switzerland), MOVAS cells were cultured in DMEM supplemented 10% (v/v) FBS (both from Gibco, Scotland, UK). Cultured were maintained at 37 °C in a humidified atmosphere of 5% CO₂—95% air. When cells achieved 90% confluence the medium was changed and cells were treated with 100 μM deuterated Oleic acid (OAd34, Sigma Aldrich) or deuterated arachidonic acid (AAd8, 25 µM, Cayman Chemical) in the presence or absence of lipolysis inhibitor, atglistatin (50 µM), phospholipase A₂ inhibitor (AACOCF3, 10 µM, Abcam), COX-1 inhibitor (SC-560, Cayman Chemical) or COX-2 inhibitor (Dup-697, Cayman Chemical) for 4 h or 24 h. Oleic acid (Cayman Chemical) for the experiments was freshly saponified using 100 mM NaOH and BSA-conjugated (10% in DPBS, low endotoxin, fatty acid-free, suitable for cell culture, sterile-filtered; Sigma Aldrich). After 4 h or 24 h of incubation cells were fixed for 4 min with 4% paraformaldehyde. Fixed cells were washed 3 times with DPBS and stored in DPBS at 4 °C until the execution of the measurements.

Mouse vascular smooth muscle cell line (MOVAS) obtained from American Type Culture Collection (ATCC) were used for other biochemical experiments. The MOVAS cells were cultured in DMEM medium (Gibco, MA, USA) containing 10% fetal bovine serum (FBS, 10099141, Gibco, MA, USA) and 1% penicillin-streptomycin (15140122, Gibco, MA, USA).

Mouse aortic vascular smooth muscle cells (MOVAS, ACTT number:CRL-2797) and FBS were purchased from Cellcook Biotech Co., China. MOVAS were cultured in DMEM (Gibco, USA) medium containing 10% FBS, 0.2 mg/ml G418 (BioFroxx, GER), penicillin, and streptomycin (Gibco, USA, 1 ： 100) at 37°C and 5% CO₂. The cells were divided into physiological group and pathological group. The physiological group included the control group (blank group), low- and high-dose GXP groups, and atorvastatin calcium group. The pathological group included endothelin-induced ET-1 group (model group), ET-1 +  low- and high-dose GXP groups, and ET-1 + atorvastatin calcium group.