Yi Li1, Bin Du, Jia-qi Pan, De-chang Chen, Da-wei Liu. 1. Department of Critical Care Medicine, Peking Union Medical College Hospital, Chinese Medical Academy, Beijing 100730, China. billliyi@yahoo.com
Abstract
OBJECTIVE: To investigate the effect of activated protein C (APC) on inflammatory responses in human umbilical vein endothelial cells (HUVEC) stimulated with lipopolysaccharide (LPS). METHODS: The second passage of collagenase digested HUVEC was divided into the following groups: serum free medium control group (SFM control), phosphate buffer solution control group (PBS control), LPS group with final concentration of 1 microg/ml (LPS group), APC group with final concentration of 7 microg/ml, Pre-APC group (APC pretreatment for 30 min prior to LPS challenge), and Post-APC group (APC administration 30 min after LPS challenge). Supernatant was harvested at 0, 4, 8, 12 and 24 h after LPS challenge. Interleukin-6 (IL-6) and Interleukin-8 (IL-8) levels were analyzed with ELISA. Cells were harvested at 24 h after LPS challenge, and total RNA was extracted. Messenger RNA levels for IL-6 and IL-8 were semi-quantitatively determined by RT-PCR. RESULTS: Compared with control group, IL-6 and IL-8 levels steadily increased 4 to 24 h after LPS stimulation. APC treatment could increase LPS-induced IL-6 and IL-8 production. The mRNA levels of IL-6 and IL-8 exhibited a similar change. CONCLUSION: APC can further increase the level of IL-6 and IL-8 induced by LPS. The effect of these elevated cytokines is still under investigation.
OBJECTIVE: To investigate the effect of activated protein C (APC) on inflammatory responses in human umbilical vein endothelial cells (HUVEC) stimulated with lipopolysaccharide (LPS). METHODS: The second passage of collagenase digested HUVEC was divided into the following groups: serum free medium control group (SFM control), phosphate buffer solution control group (PBS control), LPS group with final concentration of 1 microg/ml (LPS group), APC group with final concentration of 7 microg/ml, Pre-APC group (APC pretreatment for 30 min prior to LPS challenge), and Post-APC group (APC administration 30 min after LPS challenge). Supernatant was harvested at 0, 4, 8, 12 and 24 h after LPS challenge. Interleukin-6 (IL-6) and Interleukin-8 (IL-8) levels were analyzed with ELISA. Cells were harvested at 24 h after LPS challenge, and total RNA was extracted. Messenger RNA levels for IL-6 and IL-8 were semi-quantitatively determined by RT-PCR. RESULTS: Compared with control group, IL-6 and IL-8 levels steadily increased 4 to 24 h after LPS stimulation. APC treatment could increase LPS-induced IL-6 and IL-8 production. The mRNA levels of IL-6 and IL-8 exhibited a similar change. CONCLUSION:APC can further increase the level of IL-6 and IL-8 induced by LPS. The effect of these elevated cytokines is still under investigation.
Authors: B White; M Schmidt; C Murphy; W Livingstone; D O'Toole; M Lawler; L O'Neill; D Kelleher; H P Schwarz; O P Smith Journal: Br J Haematol Date: 2000-07 Impact factor: 6.998
Authors: B L Warren; A Eid; P Singer; S S Pillay; P Carl; I Novak; P Chalupa; A Atherstone; I Pénzes; A Kübler; S Knaub; H O Keinecke; H Heinrichs; F Schindel; M Juers; R C Bone; S M Opal Journal: JAMA Date: 2001-10-17 Impact factor: 56.272