Up-regulation interleukin-6 and interleukin-8 by activated protein C in lipopolysaccharide-treated human umbilical vein endothelial cells |
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| Authors: | Yi Li Bin Du Jia-qi Pan De-chang Chen Da-wei Liu |
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| Institution: | (1) Department of Critical Care Medicine, Peking Union Medical College Hospital, Chinese Medical Academy, Beijing, 100730, China;(2) Department of Emergency Medicine, Peking Union Medical College Hospital, Chinese Medical Academy, Beijing, 100730, China;(3) Department of Hematology, Peking Union Medical College Hospital, Chinese Medical Academy, Beijing, 100730, China |
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| Abstract: | Objective To investigate the effect of activated protein C (APC) on inflammatory responses in human umbilical vein endothelial cells
(HUVEC) stimulated with lipopoly saccharide (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 μg/ml (LPS group),
APC group with final concentration of 7 μg/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. |
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| Keywords: | Activated protein C (APC) Interleukin-6 (IL-6) Interleukin-8 (IL-8) Sepsis Human umbilical vein endothelial cell (HUVEC) |
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