Yeh Chiu-Li, Hsu Chun-Sen, Chen Soul-Chin, Hou Yu-Chen, Chiu Wan-Chun, Yeh Sung-Ling
Institute of Nutrition and Health Sciences, Taipei Medical University, Taipei, Taiwan.
Shock. 2007 Jul;28(1):39-44. doi: 10.1097/shk.0b013e31802f0190.
This study investigated the effect of different arginine (Arg) concentrations on adhesion molecule expression on endothelial cells (ECs) and leukocytes and the transendothelial migration of polymorphonuclear neutrophils (PMNs) through ECs stimulated by plasma or peritoneal drain fluid (PDF) from surgical patients. Human umbilical vein ECs (HUVECs) and PMNs from healthy subjects were treated with different concentrations (0, 50, 100, and 1000 micromol/L) of Arg for 24 h. After that, HUVECs were stimulated for 3 h with plasma or PDF from patients who underwent abdominal surgery, and PMNs were allowed to transmigrate through ECs for 2 h. The HUVEC expression of cellular adhesion molecules (CAMs) and integrin (CD11b) and the interleukin (IL) 8 receptor expression on PMNs were measured by flow cytometry. The PMNs transmigrating through ECs were also analyzed. The results showed that CAM and integrin expressions in PDF groups were higher than those in control groups. Among the PDF groups, IL-8 secretions from ECs and PMNs were lower with 100 and 1000 micromol/L Arg than with 0 and 50 micromol/L Arg, and this was consistent with the expression of the IL-8 receptor on PMNs. In addition, CAM expressions on ECs and CD11b expression on PMNs, as well as PMN transmigration, were lower with 100 and 1000 micromol/L Arg than with 0 and 50 micromol/L Arg. The HUVECs stimulated by plasma from surgical patients had similar effects on surface molecule expression as PDF; however, as shown in PDF stimulation, the effects were not so obvious. Inhibition of nitric oxide production results in high CAM and IL-8 expressions comparable with groups with low Arg administration. The results of this in vitro study suggest that ECs and PMNs were activated after patients' plasma or PDF stimulation. A low Arg concentration comparable with catabolic conditions resulted in higher adhesion molecule expression and greater transendothelial migration of neutrophils. Arginine administration at levels similar to or higher than physiological concentrations reduced IL-8 and CAM expression, and PMN transmigration was also decreased after stimulation with plasma or PDF from surgical patients. Inactivation of NO results in high CAM and IL-8 expression. This finding indicated that NO may be an important endogenous inhibitor for EC-PMN interaction and neutrophil transmigration.
本研究调查了不同浓度精氨酸(Arg)对内皮细胞(ECs)和白细胞上黏附分子表达的影响,以及多形核中性粒细胞(PMNs)通过手术患者血浆或腹腔引流液(PDF)刺激的ECs进行跨内皮迁移的情况。将来自健康受试者的人脐静脉内皮细胞(HUVECs)和PMNs用不同浓度(0、50、100和1000微摩尔/升)的Arg处理24小时。之后,用接受腹部手术患者的血浆或PDF刺激HUVECs 3小时,并让PMNs通过ECs迁移2小时。通过流式细胞术测量HUVEC上细胞黏附分子(CAMs)和整合素(CD11b)的表达以及PMNs上白细胞介素(IL)8受体的表达。还对通过ECs迁移的PMNs进行了分析。结果显示,PDF组中CAM和整合素的表达高于对照组。在PDF组中,与0和50微摩尔/升Arg相比,100和1000微摩尔/升Arg处理的ECs和PMNs分泌的IL-8较低,这与PMNs上IL-8受体的表达一致。此外,与0和50微摩尔/升Arg相比,100和1000微摩尔/升Arg处理的ECs上的CAM表达、PMNs上的CD11b表达以及PMN迁移均较低。手术患者血浆刺激的HUVECs对表面分子表达的影响与PDF相似;然而,如PDF刺激所示,其影响不那么明显。抑制一氧化氮产生会导致与低Arg给药组相当的高CAM和IL-8表达。这项体外研究结果表明,患者血浆或PDF刺激后ECs和PMNs被激活。与分解代谢状态相当的低Arg浓度导致更高的黏附分子表达和更大的中性粒细胞跨内皮迁移。给予与生理浓度相似或更高水平的精氨酸会降低IL-8和CAM表达,并且在手术患者血浆或PDF刺激后PMN迁移也会减少。一氧化氮失活会导致高CAM和IL-8表达。这一发现表明,一氧化氮可能是EC-PMN相互作用和中性粒细胞迁移的重要内源性抑制剂。