本文關(guān)鍵詞： 大麻素受體 人類中性粒細(xì)胞 細(xì)胞遷移　出處：《首都醫(yī)科大學(xué)學(xué)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(xì)胞(d HL60)遷移的影響。方法體外培養(yǎng)人早幼粒白血病細(xì)胞系HL60,使用二甲基亞砜(dimethylsulphoxide,DMSO)誘導(dǎo)為類中性粒細(xì)胞(d HL60),運用實時熒光定量聚合酶鏈反應(yīng)檢測其分化標(biāo)志物CD11b mRNA的表達(dá);應(yīng)用瓊脂糖凝膠電泳、Western blotting法及免疫熒光技術(shù)檢測其大麻素受體1(CB1)及受體2(CB2)的表達(dá);ACEA、AM281分別為CB1的藥理學(xué)激動劑和拮抗劑,JWH133、AM630分別為CB2的藥理學(xué)激動劑和拮抗劑,應(yīng)用Boyden chamber法檢測ACEA和JWH133對d HL60遷移的影響,并從藥理學(xué)阻斷CB1、CB2后,檢測其遷移功能的變化;使用鬼筆環(huán)肽染細(xì)胞肌動蛋白纖維,并應(yīng)用高內(nèi)涵掃描分析的方法對肌動蛋白纖維的聚合進(jìn)行分析。結(jié)果 d HL60在mRNA和蛋白質(zhì)水平上均表達(dá)CB1、CB2;ACEA能夠誘導(dǎo)d HL60的遷移及其細(xì)胞骨架的聚合,且其所誘導(dǎo)的遷移能夠被CB1的藥理學(xué)阻斷劑AM281所阻斷,而CB2的藥理學(xué)阻斷劑AM630對ACEA所誘導(dǎo)的遷移并無影響;給予CB2的激動劑JWH133對d HL60的遷移及細(xì)胞骨架的聚合無明顯作用。結(jié)論激活CB1能夠促進(jìn)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(xué)細(xì)胞生物學(xué)系‘肝臟保護(hù)與再生調(diào)節(jié)’北京市重點實驗室;
【基金】：國家自然科學(xué)基金(81430013) 北京市屬高等學(xué)校創(chuàng)新團(tuán)隊建設(shè)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(xì)胞屬于天然免疫的第一道防線,是人體內(nèi)最主要的固有免疫細(xì)胞之一,主要來源于骨髓。當(dāng)機體受到刺激發(fā)生炎性反應(yīng)時,中性粒細(xì)胞被激活并進(jìn)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(dāng)其遷移到炎性反應(yīng)部位,可以通過吞噬、脫顆粒作用及中性粒胞外誘捕網(wǎng)來發(fā)

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