大麻素受体1介导人类中性粒细胞dHL60的迁移功能

发布时间：2018-01-24 01:46

本文关键词： 大麻素受体人类中性粒细胞细胞迁移　出处：《首都医科大学学报》2017年03期 　论文类型：期刊论文

【摘要】：目的研究大麻素受体(cannabinoid receptors,CBs)对人类中性粒细胞(d HL60)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(d HL60),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d HL60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 d HL60在mRNA和蛋白质水平上均表达CB1、CB2;ACEA能够诱导d HL60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d HL60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d HL60的迁移。
[Abstract]:Objective to study cannabinoid receptors of cannabinoid. Methods Human promyelocytic leukemia cell line HL60 was cultured in vitro. Dimethylsulphoxide (DMSO) was used to induce neutrophil like granulocyte (HL60). The expression of CD11b mRNA was detected by real-time fluorescence quantitative polymerase chain reaction. The expression of cannabinoid receptor 1 (CB1) and receptor 2 (CB2) were detected by agarose gel electrophoresis (blotting) and immunofluorescence technique. ACEAN AM281 is the pharmacological agonist and antagonist of CB1 and JWH133AAM630 is the pharmacological agonist and antagonist of CB2, respectively. The effect of ACEA and JWH133 on the migration of d HL60 was detected by Boyden chamber method. The cell actin fibers were stained with ghoup cyclopeptide. The polymerization of actin fibers was analyzed by high intension scanning analysis. Results d HL60 expressed CB1 and CB2 on both mRNA and protein levels. ACEA could induce the migration of d HL60 and the aggregation of cytoskeleton, and the migration could be blocked by AM281, the pharmacological blocker of CB1. AM630, the pharmacological blocker of CB2, had no effect on the migration induced by ACEA. JWH133, an agonist of CB2, had no effect on the migration of d HL60 and cytoskeleton polymerization. Conclusion Activation of CB1 can promote the migration of d HL60.
【作者单位】：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;
【基金】：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)~~
【分类号】：R392
【正文快照】： 中性粒细胞属于天然免疫的第一道防线,是人体内最主要的固有免疫细胞之一,主要来源于骨髓。当机体受到刺激发生炎性反应时,中性粒细胞被激活并进入外周循环系统,后将会朝着趋化梯度区域的上游迁移[1]。当其迁移到炎性反应部位,可以通过吞噬、脱颗粒作用及中性粒胞外诱捕网来发

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