Effects of anesthetics and shear stress on the expression of E-selectin in cytokine-stimulated endothelial cells.

2000 Fiscal Year Final Research Report Summary

• Project/Area Number: 11671486
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Anesthesiology/Resuscitation studies
• Research Institution: TOYAMA MEDICAL AND PHARMACEUTICAL UNIVERSITY
• Principal Investigator: SHAKUNAGA Kiyoshi TOYAMA MEDICAL AND PHARMACEUTICAL UNIVERSITY Faculty of Medicine Research Associate, 医学部, 助手 (40187498)
• Project Period (FY): 1999 – 2000
• Keywords: adhesion molecules / E-selectin / soluble E-selectin / interleukin 1 / intravenous anesthetics / ketamine / propofol / endothelial cells

Research Abstract

INTRODUCTION : Cell-surface E-selectin is a mediator of the rolling attachment of leukocytes to the endothelium, an essential step in extravasation of leukocytes at the site of inflammation, thereby playing a key role in localized inflammatory response. Elevated levels of soluble E-selectin in serum have been reported in a variety of pathological conditions. But there are no reports about the direct effect of anesthetics on soluble E-selectin production. The aim of this study was to examine the effects of anesthetics on soluble E-selectin release and E-selectin expression in cultured human umbilical vein endothelial cells (HUVECs).
METHODS : Confluent HUVECs were preincubated with ketamine (0,10,100μM) at 37℃ for 2 h and then the confluent monolayer cells were further incubated for 4 h in the absence or the presence of IL-1 (5U/ml). After incubation, the culture medium was collected, centrifuged to remove cell debris, and used as a condtioned medium (CM). The concentrations of soluble E -Selectin in the CM were determined with a sandwich enzyme-linked immunosorbent assay (ELISA). Furthermore, we assessed E-selectin mRNA levels by Northern blottiong.
RESULTS : In the absence of ketamine, treatment with IL-1 caused a significant (p<0.05) increase in the level of soluble E-Selectin. In contrast, pretreatment of ketamine significantly reduced the stimulatory effect of IL-1. About 70% of the secretion was inhibited by 10μM ketamine. In the absence of IL-1, ketamine treatment did not cause a significant change in its soluble E-Selectin value. The mRNA encoding E-selectin was expressed much more slowly and to a lesser extent under the presence of ketamine.
DISCUSSION : These results suggest that ketamine suppresses the production and release of soluble E-Selectin from IL-1 stimulated HUVECs. at least in part by down-regulation of the E-Selectin mRNA expression. Our study suggests that ketamine could be used in patients with the systemic inflammatory response syndrome through its inhibitory effect on soluble E-Selectin production.