Human pulmonary microvascular endothelial cells (HPMECs) were plated at the density of 5 x 105 cells/ml on either culture dishes or collagen I-coated flexible bottom BioFlex plates in endothelial cell medium with 5% fetal bovine serum, 1% endothelial cell growth factor, and 1% penicillin/streptomycin solution.
The confluent HPMECs stably transfected with the FSTL-1 siRNA, or treated for 2 h with 10 [micro]mol/L of the NLRP3 inhibitor MCC950, were exposed to cyclic stretch using a FX-5000T Flexercell Tension Plus system (Flexcell International, McKeesport, PA, USA) equipped with a 25-mm BioFlex loading station.
In the present study, we used HPMECs to study the underlying inflammatory mechanisms of VILI.
HPMECs were maintained in Dulbecco's modified Eagle's medium (DMEM)/F12 containing 10% (v/v) fetal bovine serum at 37[degrees]C under a humidified atmosphere of 5% C[O.sub.2] and 95% air.
HPMECs were seeded in 6-well plates at a density of 3 x [10.sup.5] cells per well in 2 mL antibiotic-free M199 medium with 10% FBS added.
Glucocorticoid receptor alpha (GR[alpha]) and I[kappa]B[alpha] expression in LPS-stimulated HPMECs with methylprednisolone
Bringing this in was quite a battle and I was quite all in when the fish was safe on board" (HPMEC
Human lung fibroblasts (HLF-1), human pulmonary microvascular endothelial cells (HPMECs) and human type II alveolar epithelial cells (A549 cell line) were purchased from the cell bank of the Chinese Academy of Sciences.
HPMECs were maintained in DMEM (high glucose) containing 10% (v/v) fetal bovine serum, 100kU/l penicillin and 100mg/l streptomycin at 37 [degrees]C in a humidified 5% C[0.sub.2].
HPMECs were cultured in vitro during the hypoxic condition
We chose 4 concentrations of PHC to determine the optimal dose of PHC on [beta]-arrestin-1 expression in LPS-induced HPMEC. It was demonstrated that 2 [micro]g/mL PHC upregulated expression of [beta]-arrestin-1 most significantly (Figure 4).
Then, the time point of PHC on [beta]-arrestin-1 expression in LPS-induced HPMEC was observed.