Effects Of Ketamine On The Expression Of ICAM-1,NF-κB,L-selectin And β₂-integrin

Objective: (1)To investigate the effect of ketamine on the expression of intercellular adhesive molecule-l(ICAM-l) and the translocation of nuclear kappa B into nuclei of Human umbilical venous endothelial cells (HUVECs); (2)To investigate the effect of ketamine on the expression of L-selectin and β₂-integrin of neutrophils by formyl-methi-onyl-leucy-phenylalanine(FMLP). Methods:There were two section in the experiment.1. HUVECs are isolated by pancreatic enzyme and cultured in DMEM culture medium at 37℃ 5%CO₂ incubator when a monolayer of endothelial cells has grown. (l)Effect of ketamine on the expression of ICAM-1 of HUVECs: The cultured HUVECs were stimulated with lipopolysaccharide 1000ng/ml for 24 hours in the absence or presence of ketamine (0. 3μg/ml 3. 0μg/ml 30μg/ml 300μg/ml). Subsequently, the expression of ICAM-1 were obtained by flow cytometric analysis. (2)Effect of ketamine on the translocation of nuclear factor kappa B into nuclei: The cultured HUVECs were stimulated with lipopolysaccharide 1000ng/ml for 2 hours in the absence or presence of ketamine (0. 3μg/ml 3.0μg/ml 30μg/ml 300μg/ml). The translocation of nuclear factor kappa B into nuclei are stained with immunohistochemistry technique. 2. Effect of ketamine on the expression of L- selectin and β₂-integrin of neutrophils : Neutrophils were incubated with FMLP 1.0μmol/L for 20min in the absence or presence of ketamine (0. 3 μg/ml 3.0μg/ml 30μg/ml 300μg/ml). Subsequently, the expression of them were obtained by flow cytometric analysis. Results: Not stimulated , theexpression of ICAM-1 of the HUVECs were very less (average desity of flurescence:1. 34+0. 24). Ketamine(3. 0μg/ml,30μg/ml,300 μg/ml) inhibited the expression of ICAM-1 and translocation of nuclear factor kappa B into nuclei. There was relationship of positive correlation between ICAM-1 and nuclear factor kappa B . Ketamine also repressed FMLP-induced up-regulated of β₂-integrin and shedding of L-selectin , in a concentration-dependent manner. Conclusion:Lipopolysaccharide can induce up-regulated expression of ICAM-1 of HUVECs. However , ketamine can inhibit the expression of ICAM-1 and the translocation of nuclear factor kappa B into nuclei of human umbilical vein endothelial cells with lipopolysaccharide , and restrain FMLP-induced the activation of neutrophils .