A preliminary in vitro study on the activation of polymorphonuclear cells and vascular endothelial cells co-cultured with and stimulated by lipopolysaccharide.
- Author:
Jianming LI
1
;
Qian CAI
;
Hong ZHOU
;
Guangxia XIAO
Author Information
- Publication Type:Journal Article
- MeSH: Adult; Cell Line; Cells, Cultured; Coculture Techniques; Dose-Response Relationship, Drug; Endothelium, Vascular; cytology; drug effects; metabolism; Humans; Interleukin-6; metabolism; Lipopolysaccharides; pharmacology; Neutrophils; cytology; drug effects; metabolism; Time Factors; Tumor Necrosis Factor-alpha; metabolism
- From: Chinese Journal of Burns 2002;18(2):78-80
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo explore the activation of polymorphonuclear cells (PMNs) and vascular endothelial cells co-cultured with and stimulated by lipopolysaccharide.
METHODSPMNs in concentration of 2 x 10(6)/ml isolated from healthy volunteers by Percoll gradient were added to monolayer of ECV-304 cells grown to confluency, then different groups were prepared according to final concentration of lipopolysaccharide. The morphological change was observed under invert microscope. The changes in TNFalpha and IL-6 levels of the supernatant of the cultured cells were determined at 4, 8, 12 and 24 hours after culturation.
RESULTSThe TNFalpha production of cultured pure ECV-304 exhibited no remarkable change when stimulated by different concentrations of LPS. But the TNFalpha production of the ECV-304 increased significantly when co-cultured with PMNs at 4 hr and stimulated by LPS in concentration of 10 micro g/ml, and increased at 8 hours and lasted up to 24 hours of culturation in higher levels (P < 0.05). The IL-6 production of cultured pure ECV-304 increased obviously along with the increase of LPS concentration, and it showed no change when PMNs co-cultured with ECV-304. While the IL-6 level in the supernatant of co-cultured ECV-304 with PMNs increased sharply when stimulated by both low (100 ng/ml) and high (1 micro g/ml) concentrations of LPS and maintained at high levels up to 24 hours of culturation. The higher the concentration of LPS was, the quicker the IL-6 level increased.
CONCLUSIONCo-cultured PMNs and endothelial cells could be activated and activation state could be maitained by low concentrations of LPS.