大麻素受体1介导人类中性粒细胞dHL60的迁移功能

doi:10.3969/j.issn.1006-7795.2017.03.018

首都医科大学学报 ›› 2017, Vol. 38 ›› Issue (3): 417-422.doi: 10.3969/j.issn.1006-7795.2017.03.018

大麻素受体1介导人类中性粒细胞dHL60的迁移功能

樊晓婷, 田蕾, 杨琳, 李丽英

首都医科大学细胞生物学系‘肝脏保护与再生调节’北京市重点实验室, 北京 100069

收稿日期:2016-08-05 出版日期:2017-05-21 发布日期:2017-06-14
通讯作者: 李丽英 E-mail:liliying@ccmu.edu.cn
基金资助:
国家自然科学基金（81430013），北京市属高等学校创新团队建设与教师职业发展计划项目（IDHT20150502）。

Cannabinoid receptor 1 mediated the migration of human neutrophil-like cell line dHL60

Fan Xiaoting, Tian Lei, Yang Lin, Li Liying

Department of Cell Biology, Municipal Laboratory for Liver Protection and Regulation of Regeneration, Capital Medical University, Beijing 100069, China

Received:2016-08-05 Online:2017-05-21 Published:2017-06-14
Supported by:
This study was supported by National Natural and Science Foundation of China (81430013), Project of Construction of Innovative Teams and Teacher Career Development for Universities and Colleges Under Beijing Municipality (IDHT20150502).

摘要/Abstract

摘要： 目的研究大麻素受体（cannabinoid receptors，CBs）对人类中性粒细胞（dHL60）迁移的影响。方法体外培养人早幼粒白血病细胞系HL60，使用二甲基亚砜（dimethylsulphoxide，DMSO）诱导为类中性粒细胞（dHL60），运用实时荧光定量聚合酶链反应检测其分化标志物CD11b mRNA的表达；应用琼脂糖凝胶电泳、Western blotting法及免疫荧光技术检测其大麻素受体1（CB1）及受体2（CB2）的表达；ACEA、AM281分别为CB1的药理学激动剂和拮抗剂，JWH133、AM630分别为CB2的药理学激动剂和拮抗剂，应用Boyden chamber法检测ACEA和JWH133对dHL60迁移的影响，并从药理学阻断CB1、CB2后，检测其迁移功能的变化；使用鬼笔环肽染细胞肌动蛋白纤维，并应用高内涵扫描分析的方法对肌动蛋白纤维的聚合进行分析。结果 dHL60在mRNA和蛋白质水平上均表达CB1、CB2；ACEA能够诱导dHL60的迁移及其细胞骨架的聚合，且其所诱导的迁移能够被CB1的药理学阻断剂AM281所阻断，而CB2的药理学阻断剂AM630对ACEA所诱导的迁移并无影响；给予CB2的激动剂JWH133对dHL60的迁移及细胞骨架的聚合无明显作用。结论激活CB1能够促进dHL60的迁移。

关键词: 大麻素受体, 人类中性粒细胞, 细胞迁移

Abstract: Objective To investigate the effect of cannabinoid receptors (CBs) on migration of human neutrophil-like cell line dHL60. Methods HL60 cell was cultivated in vitro and stimulated by dimethylsulphoxide (DMSO) to differentiate into neutrophil-like cells (dHL60), RT-PCR was used to detect the CD11b mRNA expression as a differentiation marker. Western blotting and agarose gel electrophoresis were used to detect the expression of CB1 and CB2. Boyden chamber was used for cell migration assay. Filamentous actin (F-actin) stained with phalloidin was examined by high content analysis. Results In this study, we present evidence that dHL60 significantly expressed CB1 and CB2; the treatment of ACEA (agonist of CB1) caused an increase in dHL60 migration and F-actin remodeling, the pharmacological inhibition of CB1 markedly attenuated ACEA-induced migration; the treatment of JWH133 (agonist of CB2) has no effect on migration and F-actin remodeling. Conclusion CB1 mediated the migration of human neutrophil-like cell line dHL60.

Key words: cannabinoid receptors(CBs), human neutrophil-like cell, cell migration

中图分类号:

樊晓婷, 田蕾, 杨琳, 李丽英. 大麻素受体1介导人类中性粒细胞dHL60的迁移功能[J]. 首都医科大学学报, 2017, 38(3): 417-422.

Fan Xiaoting, Tian Lei, Yang Lin, Li Liying. Cannabinoid receptor 1 mediated the migration of human neutrophil-like cell line dHL60[J]. Journal of Capital Medical University, 2017, 38(3): 417-422.

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大麻素受体1介导人类中性粒细胞dHL60的迁移功能

Cannabinoid receptor 1 mediated the migration of human neutrophil-like cell line dHL60

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