本文關鍵詞： 大麻素受體 人類中性粒細胞 細胞遷移　出處：《首都醫(yī)科大學學報》2017年03期 　論文類型：期刊論文

【摘要】：目的研究大麻素受體(cannabinoid receptors,CBs)對人類中性粒細胞(d HL60)遷移的影響。方法體外培養(yǎng)人早幼粒白血病細胞系HL60,使用二甲基亞砜(dimethylsulphoxide,DMSO)誘導為類中性粒細胞(d HL60),運用實時熒光定量聚合酶鏈反應檢測其分化標志物CD11b mRNA的表達;應用瓊脂糖凝膠電泳、Western blotting法及免疫熒光技術檢測其大麻素受體1(CB1)及受體2(CB2)的表達;ACEA、AM281分別為CB1的藥理學激動劑和拮抗劑,JWH133、AM630分別為CB2的藥理學激動劑和拮抗劑,應用Boyden chamber法檢測ACEA和JWH133對d HL60遷移的影響,并從藥理學阻斷CB1、CB2后,檢測其遷移功能的變化;使用鬼筆環(huán)肽染細胞肌動蛋白纖維,并應用高內涵掃描分析的方法對肌動蛋白纖維的聚合進行分析。結果 d HL60在mRNA和蛋白質水平上均表達CB1、CB2;ACEA能夠誘導d HL60的遷移及其細胞骨架的聚合,且其所誘導的遷移能夠被CB1的藥理學阻斷劑AM281所阻斷,而CB2的藥理學阻斷劑AM630對ACEA所誘導的遷移并無影響;給予CB2的激動劑JWH133對d HL60的遷移及細胞骨架的聚合無明顯作用。結論激活CB1能夠促進d HL60的遷移。
[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.
【作者單位】： 首都醫(yī)科大學細胞生物學系‘肝臟保護與再生調節(jié)’北京市重點實驗室;
【基金】：國家自然科學基金(81430013) 北京市屬高等學校創(chuàng)新團隊建設與教師職業(yè)發(fā)展計劃項目(IDHT20150502)~~
【分類號】：R392
【正文快照】： 中性粒細胞屬于天然免疫的第一道防線,是人體內最主要的固有免疫細胞之一,主要來源于骨髓。當機體受到刺激發(fā)生炎性反應時,中性粒細胞被激活并進入外周循環(huán)系統(tǒng),后將會朝著趨化梯度區(qū)域的上游遷移[1]。當其遷移到炎性反應部位,可以通過吞噬、脫顆粒作用及中性粒胞外誘捕網來發(fā)

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