中文摘要：

本文旨在探讨I组代谢型谷氨酸受体（group I metabotropic glutamate receptor,mGluRI）在中性粒细胞上的表达,以及该受体的激活对中性粒细胞与内皮细胞相互黏附的影响。取健康人新鲜静脉血,Ficoll-Hypaque密度梯度离心法分离中性粒细胞,免疫细胞化学法和real-time PCR法检测中性粒细胞mGluRI（包括mGluR1和mGluR5）的表达,分别应用不同浓度的mGluRI特异性激动剂S-3,5-二羟基苯甘氨酸（S-3,5-dihydroxy-phenylglycine,S-DHPG）处理中性粒细胞不同时间,通过比色法检测中性粒细胞和人正常脐静脉内皮细胞（human normal umbilical vein endothelial cells,HUVE-12）黏附率,采用流式细胞术测定中性粒细胞黏附分子CD11a表达的变化。结果证实中性粒细胞表达mGluRI（mGluR1/5）;S-DHPG在1×10-8~1×10-6mol/L浓度范围内呈剂量依赖性地提高中性粒细胞与内皮细胞之间的黏附率（P〈0.05或P〈0.01）;1×10-6mol/L S-DHPG单独作用于中性粒细胞0.5h,即可促进中性粒细胞黏附于内皮细胞（P〈0.01）,但没有时间依赖性;1×10-6mol/L S-DHPG单独作用于中性粒细胞可促进CD11a表达（P〈0.01）;mGluRI拮抗剂（RS）-α-甲基-4-羧基苯甘氨酸[（RS）-α-methyl-4-carboxyphenylglycine,（±）-MCPG]（0.5mmol/L）可以显著阻断激动剂S-DHPG（1×10-6mol/L,1h）的促黏附效应（P〈0.01）。上述结果证实了中性粒细胞膜上mGluRI的激活可增强中性粒细胞表面黏附分子CD11a的表达,促进中性粒细胞与内皮细胞的黏附。

英文摘要：

L-glutamate （Glu） is an excitatory neurotransmitter in the mammalian central nervous system. Relatively much attention has been paid to functional expression of Glu signaling molecules in peripheral tissues very recently. The present study tested the hypothesis that the activation of group I metabotropic glutamate receptor （mGluRI） in neutrophils stimulated neutrophils adherence to endothelial cells by increasing the surface expression of certain adhesion molecules. Peripheral blood was obtained by venipuncture from healthy donors, and the neutrophils were isolated by Ficoll-Hypaque gradient centrifugation. Neutrophils floating into DMEM/ F12 culture medium containing 10% fetal bovine serum were then used immediately. Immunocytochemistry and real-time quantitative RT-PCR were used to detect the expression of mGluRI （mGluR1 and mGluR5） in neutrophils. The adherence of neutrophils to cultured human normal umbilical vein endothelial cells （HUVE-12） was measured by the c01orimetric method. Cell surface expression of adhesion molecule CD 11 a in the neutrophils was determined by flow cytometry. Immunocytochemistry and real-time quantitative RT- PCR showed that mGluR1 and mGluR5 were constitutively expressed in neutrophils. Application of mGluRI agonist S-3,5-dihydroxyphenylglycine （S-DHPG） （1× 10^-8-1× 10^-6 mol/L） showed a dose-dependent stimulatory effect on the adherence of neutro-. phils to HUVE-12 （P〈0.05 or P〈0.01）, with a maximum effect at 1× 10^-6 mol/L （P〈0.01）. Incubations as short as 30rain were sufficient to induce increased adherence after the beginning of S-DHPG treatment. Following time extension （0：5-5 h）, S-DHPG （1 × 10^-6mol/L） increased the rate ofneutrophils adhesion to HUVE-12 with a maximum effect at 0.5 h （P〈0.01）. However, a time-dependent effect of S-DHPG on the rate of neutrophils adhesion to HUVE-12 was not observed during the experimental period. 1 × 10^-6 mol/L of S-DHPG also induced an increased surface expression of adhesion mole