Effects of peripheral cannabinoid receptor ligands on motility and polarization in neutrophil-like HL60 cells and human neutrophils
Effects of peripheral cannabinoid receptor ligands on motility and polarization in neutrophil-like HL60 cells and human neutrophils
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DOI：
10.1074/jbc.m510871200
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发表时间：
2006-05-05
影响因子：
4.8
通讯作者：
Yamamura, H
中科院分区：
文献类型：
--
作者：
Kurihara, R;Tohyama, Y;Yamamura, H
通过使用分化为中性粒细胞样细胞的人早幼粒细胞 HL60 细胞和从全血中分离的人中性粒细胞，研究了外周大麻素受体（CB2）在中性粒细胞迁移中的可能作用。通过流式细胞术证实了 HL60 细胞、嗜中性粒细胞样 HL60 细胞和人嗜中性粒细胞上 CB2 的细胞表面表达。在用 CB2 配体 JWH015 和 2-花生四烯酰甘油 (2-AG) 刺激后，中性粒细胞样 HL60 细胞在不同方向上快速伸展和缩回一个或多个含有 F-肌动蛋白的伪足，而不是形成迁移白细胞通常表现出的前/后极性。 Rho-GTPase RhoA 的活性响应 CB2 刺激而降低，而 Rac1、Rac2 和 Cdc42 活性则增加。此外，用 RhoA 依赖性蛋白激酶 (p160-ROCK) 抑制剂 Y27632 处理细胞产生与 CB2 刺激细胞相似的细胞骨架组织。在人类中性粒细胞中，JWH015 和 2-AG 均未诱导运动或形态改变。然而，用这些配体预处理中性粒细胞会破坏 N-甲酰基-L-甲硫氨酰-L-亮氨酰-L-苯丙氨酸 (fMLP) 诱导的前/后极化和迁移，并且还显着抑制 fMLP 诱导的 RhoA 活性。这些结果表明CB2可能通过控制RhoA激活来调节过度炎症反应，从而抑制中性粒细胞迁移。
The possible role of the peripheral cannabinoid receptor (CB2) in neutrophil migration was investigated by using human promyelocytic HL60 cells differentiated into neutrophil-like cells and human neutrophils isolated from whole blood. Cell surface expression of CB2 on HL60 cells, on neutrophil-like HL60 cells, and on human neutrophils was confirmed by flow cytometry. Upon stimulation with either of the CB2 ligands JWH015 and 2-arachidonoylglycerol (2-AG), neutrophil-like HL60 cells rapidly extended and retracted one or more pseudopods containing F-actin in different directions instead of developing front/rear polarity typically exhibited by migrating leukocytes. Activity of the Rho-GTPase RhoA decreased in response to CB2 stimulation, whereas Rac1, Rac2, and Cdc42 activity increased. Moreover, treatment of cells with RhoA-dependent protein kinase (p160-ROCK) inhibitor Y27632 yielded cytoskeletal organization similar to that of CB2-stimulated cells. In human neutrophils, neither JWH015 nor 2-AG induced motility or morphologic alterations. However, pretreatment of neutrophils with these ligands disrupted N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP)-induced front/rear polarization and migration and also substantially suppressed fMLP-induced RhoA activity. These results suggest that CB2 might play a role in regulating excessive inflammatory response by controlling RhoA activation, thereby suppressing neutrophil migration.