Interleukin-8 production from human umbilical vein endothelial cells during brief hyperglycemia : The effect of tumor necrotic factor alpha

背景。本研究评估了不同高血糖状态下内皮细胞产生趋化因子白细胞介素(IL)-8的变化，并研究了急性炎症反应相关的高血糖是否会促进内皮细胞产生IL-8。材料与方法。人脐内皮细胞（HUVECs）以1×105细胞/孔的浓度播种并培养。用含有不同浓度葡萄糖（培养基最终葡萄糖浓度分别为100、200、300、400、500 mg/dL， n= 7）和不含100 ng tnf - α的培养基199代替。37℃下12或24 h后，收集培养物上清，80℃下保存至细胞因子检测。结果高糖培养下HUVECs的IL-8产量显著高于对照培养（葡萄糖浓度为100 mg/dL） （P<0.05）。此外，与对照组相比，在所有葡萄糖浓度和两种时间过程中培养的HUVECs中，与tnf - α升高相关的高血糖可提高IL-8的产生水平（P<0.05）。在本研究中，我们观察到内皮细胞在短期高血糖期间产生IL-8的显著增加，并且通过TNF治疗获得了类似但显著更强的增加。这些发现提示，创伤后高血糖伴急性炎症反应可能是继发性组织损伤的高危人群。

Background.This study evaluated the changes in chemokine interleukin (IL)-8 production from endothelial cells under various hyperglycemic conditions and investigated whether the hyperglycemia associated with the acute inflammatory response could enhance the IL-8 production from the endothelial cells.Materials and Methods.Human umbilical endothelial cells (HUVECs) were seeded at a concentration of 1×105 cells/well and cultured. The culture medium was replaced with Medium 199 containing various concentrations of glucose (final glucose concentration of culture medium was 100, 200, 300, 400, 500 mg/dL ; n= 7 each) with or without 100 ng of TNF-alpha. After 12 or 24 h at 37℃, the supernatants were collected from the cultures and stored at-80℃ until cytokine assay. IL-8 levels of the samples from the supernatants were quantified using a commercially available ELISA kit.Results.The IL-8 production by the HUVECs was significantly higher in the high glucose culture than in the control culture (glucose concentration of 100 mg/dL) (P<0.05). Moreover, the hyperglycemia associated with elevated TNF-alpha was found to enhance the level of IL-8 production by the HUVECs cultured at all glucose concentrations and over both time courses, compared to the control (P<0.05).Conclusions.In this study we observed a significant augmentation of IL-8 production by endothelial cells during short-term hyperglycemia, and a similar but significant stronger augmentation was obtained through TNF treatment. These findings suggest that the hyperglycemia associated with acute inflammatory response after trauma may run into the patients at high risk for secondary tissue damage.