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

简介：细胞表面E-选择素是白细胞滚动附着于内皮的介质，这是白细胞在炎症部位外渗的重要步骤，从而在局部炎症反应中起关键作用。血清中可溶性E-选择素水平升高已被报道在多种病理条件下。但麻醉药对可溶性E-选择素产生的直接影响尚未见报道。方法：体外培养的人脐静脉内皮细胞（HUVECs），分别加入氯胺酮（0、10、100 μM）37℃孵育2 h后，再加入IL-1（5 U/ml）孵育4 h，观察麻醉药对HUVECs可溶性E-选择素释放及E-选择素表达的影响。孵育后，收集培养基，离心以除去细胞碎片，并用作条件培养基（CM）。可溶性E 关于我们 - 用夹心酶联免疫吸附试验（ELISA）测定CM中的选择素。此外，我们通过北方印迹法评估E-选择素mRNA水平。结果：在不存在氯胺酮的情况下，用IL-1处理引起可溶性E-选择素水平的显著（p<0.05）增加。与此相反，氯胺酮的预处理显著降低了IL-1的刺激作用。10μM氯胺酮可抑制约70%的分泌。在不存在IL-1的情况下，氯胺酮处理未引起其可溶性E-选择素值的显著变化。编码E-选择素的mRNA表达得更慢，并在较小的程度下存在ketamine.DISCUSSION：这些结果表明，氯胺酮抑制生产和释放可溶性E-选择素从IL-1刺激的HUVECs。至少部分通过下调E-选择素mRNA表达。我们的研究表明，氯胺酮可以通过抑制可溶性E-选择素的产生来治疗全身炎症反应综合征患者。少

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