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

麻醉剂和剪切应力对细胞因子刺激的内皮细胞中 E-选择素表达的影响。

• 批准号: 11671486
• 负责人: SHAKUNAGA Kiyoshi
• 金额: $ 2.24万
• 依托单位: TOYAMA MEDICAL AND PHARMACEUTICAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671486/
• 关键词: adhesion molecules; E-selectin; soluble E-selectin; interleukin 1; intravenous anesthetics; ketamine; propofol; endothelial cells

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 … More -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. Less

前言：细胞表面E-选择素是白细胞与内皮细胞滚动附着的中介物，是炎症部位白细胞外渗的重要步骤，在局部炎症反应中起关键作用。血清中可溶性E-选择素水平的升高在各种病理条件下都有报道。但目前还没有关于麻醉药对可溶性E-选择素产生的直接影响的报道。为探讨麻醉药对体外培养的人脐静脉内皮细胞(HUVECs)可溶性E-选择素释放及E-选择素表达的影响。方法：用氯胺酮(0，10,100μM)与体外培养的人脐静脉内皮细胞(HUVECs)在37℃条件下孵育2 h，再加入IL-1(5U/ml)或不加IL-1(5U/ml)孵育4 h。培养后收集培养液，离心除去细胞碎片，作为条件培养液(CM)。可溶性E-…的浓度用双抗体夹心酶联免疫吸附试验(ELISA)检测CM中的More-Selectin。结果：在无氯胺酮的情况下，IL-1可显著增加可溶性E-选择素的水平(P<0.05)。相反，氯胺酮的预处理显著降低了IL-1的刺激作用。约70%的分泌被10μM氯胺酮抑制。在没有IL-1的情况下，氯胺酮处理对其可溶性E-选择素值没有显著影响。结论：氯胺酮抑制IL-1刺激的人脐静脉内皮细胞产生和释放可溶性E-选择素。至少部分通过下调E-选择素mRNA的表达。我们的研究提示，氯胺酮可以通过抑制可溶性E-选择素的产生而用于全身炎症反应综合征患者。较少