Heparan sulfate proteoglycans are involved in opiate receptor-mediated cell migration
Heparan sulfate proteoglycans are involved in opiate receptor-mediated cell migration
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DOI：
10.1021/bi035295i
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发表时间：
2004-01-13
期刊：
影响因子：
2.9
通讯作者：
Wiedermann, CJ
中科院分区：
文献类型：
--
作者：
Kaneider, NC;Dunzendorfer, S;Wiedermann, CJ
阿片受体在免疫系统的细胞中表达，其天然和合成配体具有强大的免疫调节作用，已有报道。在一些研究中，阿片受体拮抗剂纳洛酮本身就显示出免疫调节作用。我们研究了纳洛酮对白细胞趋化作用的影响。使用改良的Boyden小室在微孔滤膜分析中测试细胞迁移，并使用放射性标记结合分析来研究受体的表达。纳洛酮在纳摩尔浓度诱导外周血单核细胞和中性粒细胞趋化，并抑制其向β-内啡肽、血管紧张素II、生长抑素或白介素8的迁移，但不向RANTES、血管活性肠肽或P物质迁移。配基结合研究显示，纳洛酮对白介素8与中性粒细胞的结合无影响。从细胞表面的蛋白多聚糖上切割出的硫酸乙酰肝素完全抑制了纳洛酮的趋化和失活特性，但不能抑制其他引诱剂。用肝素酶、软骨素酶、氯酸钠和抗Syndecan-4抗体处理细胞后，趋化特性被取消，表明Syndecan-4参与其中。向纳洛酮迁移的程度可通过二甲基鞘氨醇(一种特殊的鞘氨醇激酶抑制剂)进行预处理而减少。由于白细胞趋化中的Syndecan-4信号涉及鞘氨醇激酶的激活，结果表明纳洛酮在细胞迁移中与Syndecan-4功能相互作用，并提示硫酸乙酰肝素蛋白多糖作为Delta-阿片受体家族成员的辅助受体发挥作用。
Opioid receptors are expressed in cells of the immune system, and potent immunomodulatory effects of their natural and synthetic ligands have been reported. In some studies, the opiate receptor antagonist naloxone itself displayed immunomodulatory actions. We investigated effects of naloxone on leukocyte chemotaxis. Cell migration was tested in micropore filter assays using modified Boyden chambers, and receptor expression was investigated using radiolabel binding assays. Naloxone induced peripheral blood nonadherent mononuclear cell and neutrophil chemotaxis at nanomolar concentrations and deactivated their migration toward beta-endorphin, angiotensin II, somatostatin, or interleukin-8 but not toward RANTES, vasoactive intestinal peptide, or substance P. Ligand binding studies showed no alteration in the binding of interleukin-8 to neutrophils by naloxone. Cleavage of heparan sulfate from proteoglycans on the cells' surface completely inhibited chemotactic and deactivating properties of naloxone but not other attractants. Chemotactic properties were abolished by pretreating cells with heparinase, chondroitinase, sodium chlorate, and anti-syndecan-4 antibodies, indicating the involvement of syndecan-4. The extent of migration toward naloxone was diminished by pretreatment with dimethylsphingosine, a specific sphingosine kinase inhibitor. As syndecan-4 signaling in leukocyte chemotaxis involves activation of sphingosine kinase, results indicate that naloxone interacts with syndecan-4 function in cell migration and suggest a role for heparan sulfate proteoglycans as coreceptors to members of the delta-opiate receptor family.