Schaefer U, Schneider A, Rixen D, Neugebauer E
Department of Surgery, University of Cologne, Germany.
Inflamm Res. 1998 Jun;47(6):256-64. doi: 10.1007/s000110050327.
In order to understand the underlying mechanism of histamine stimulated inflammatory responses, histamine receptor subtypes and signal transduction pathways by which histamine mediates the stimulation of neutrophil adhesion to endothelial cells has been studied in vitro.
Human neutrophils and human umbilical vein endothelial cells.
Confluent human endothelial cell layer were incubated with histamine (1 mM), H1, H2 or H3 receptor agonists: fluorophenylhistamine (10 microM), amthamine (10 microm), methylhistamine (10 microM), respectively. Ten minutes prior to histamine (1 mM) stimulation H1, H2 or H3 receptor antagonists (dimethindene, 100 microM; famotidine, 1OO microM, thioperamid 100 microM, respectively) were added. Histamine stimulated signal transduction pathways were inhibited by adding phospholipase C inhibitor 2-nitro-4-carboxyphenyl N,N-diphenylcarbamat (200 microM), adenylate cyclase inhibitor 9-(2 tetrahydrofuryl)adenine (80 microM), nitric oxide synthase isozymes inhibitor S-ethylisothiourea (1 microM) or guanylate cyclase inhibitor (LY 83583; 10 microM). Neutrophil adhesion was monitored at 30, 60, 90, 120, 150, 180 and 210 min.
Neutrophil adhesion to endothelial cells was quantified by analysing alkaline phosphatase activity.
Histamine stimulation of endothelial cells resulted in a biphasic time and concentration dependent pattern of neutrophil adhesion. This pattern of neutrophil adhesion was mimicked by stimulation of endothelial cells with H1 or H2 agonists. Stimulation of endothelial cells with an H3 agonist had no effect on neutrophil binding. Inhibition of phospholipase C (PLC), nitric oxide synthase isozymes (NOS) or guanylate cyclase (GC) resulted in a significant decrease of neutrophil binding to histamine or agonist stimulated endothelial cells. An increase of neutrophil binding to unstimulated or to agonist stimulated endothelial cells was observed during inhibition of adenylate cyclase.
Our results suggest that histamine stimulated neutrophil adhesion is due to H1 and H2 receptor mediated activation of PLC, NOS and GC. Increase of cAMP concentration seems to mediate an inhibitory effect on PMN adhesion to endothelial cells.
为了解组胺刺激炎症反应的潜在机制，体外研究了组胺介导中性粒细胞黏附于内皮细胞的组胺受体亚型及信号转导途径。
人中性粒细胞和人脐静脉内皮细胞。
将汇合的人内皮细胞层分别与组胺（1 mM）、H1、H2或H3受体激动剂：氟苯组胺（10 μM）、氨他明（10 μM）、甲基组胺（10 μM）孵育。在组胺（1 mM）刺激前10分钟，分别加入H1、H2或H3受体拮抗剂（分别为100 μM的二甲茚定、100 μM的法莫替丁、100 μM的硫丙咪胺）。通过加入磷脂酶C抑制剂2-硝基-4-羧基苯基N,N-二苯基氨基甲酸酯（200 μM）、腺苷酸环化酶抑制剂9-(2-四氢呋喃基)腺嘌呤（80 μM）、一氧化氮合酶同工酶抑制剂S-乙基异硫脲（1 μM）或鸟苷酸环化酶抑制剂（LY 83583；10 μM）抑制组胺刺激的信号转导途径。在30、60、90、120、150、180和210分钟监测中性粒细胞黏附情况。
通过分析碱性磷酸酶活性定量中性粒细胞与内皮细胞的黏附。
组胺刺激内皮细胞导致中性粒细胞黏附呈双相时间和浓度依赖性模式。H1或H2激动剂刺激内皮细胞可模拟这种中性粒细胞黏附模式。H3激动剂刺激内皮细胞对中性粒细胞结合无影响。抑制磷脂酶C（PLC）、一氧化氮合酶同工酶（NOS）或鸟苷酸环化酶（GC）导致中性粒细胞与组胺或激动剂刺激的内皮细胞的结合显著减少。在抑制腺苷酸环化酶期间，观察到中性粒细胞与未刺激或激动剂刺激的内皮细胞的结合增加。
我们的结果表明，组胺刺激的中性粒细胞黏附是由于H1和H2受体介导的PLC、NOS和GC的激活。cAMP浓度的增加似乎介导了对PMN黏附于内皮细胞的抑制作用。
