The Effects Of Activated Protein C On LPS-induced HUVEC

Objective:To study the effect of activated protein C (APC) on inflammatory, coagulatory and fibrinolytic response in human umblilical vein endothelial cell (HUVEC) stimulated with lipopolysaccharide (LPS).Method:HUVEC was isolated with collagenase digestion. When the second passage was in confluency, HUVEC was divided into the following groups: SFM control (serum free medium control group), PBS control (phosphate buffer solution control group), LPS group (with final concentration of 1 ng/ml), APC group (with final concentration of 7 μg/ml), PRE-APC group (APC pretreatment for 30 minutes prior to LPS challenge), and POST-APC group (APC administration 30 minutes after LPS challenge). Supernatant was harvested at 0, 4, 8, 12 and 24 hours after LPS challenge, t-PA, PAI-1, IL-6, and IL-8 levels were analyzed with ELISA. Cells were harvested at 24 hours after LPS challenge, and total RNA was extracted. Messenger RNA levels for IL-6, IL-8, t-PA, PAI-1, TF, EPCR, and TM were semi-quantitatively determined with RT-PCR.Results:...