本文選題：人抗原R + 大麻素受體��； 參考：《首都醫(yī)科大學(xué)學(xué)報》2016年01期

【摘要】：目的研究大麻素受體1(cannabinoid receptor 1,CB1)在單核巨噬細(xì)胞遷移中的重要作用以及RNA結(jié)合蛋白人抗原R(human antigen R,HuR)參與其中的可能機制。方法選用單核巨噬細(xì)胞系J774A.1,應(yīng)用瓊脂糖凝膠電泳和免疫熒光染色技術(shù)鑒定J774A.1中CB1以及HuR的表達(dá);ACEA和AM281分別為CB1的藥理學(xué)激動劑和拮抗劑,應(yīng)用Boyden chamber法檢測ACEA和AM281對J774A.1遷移活性的影響。HuR的基因干擾用于確定激活CB1誘導(dǎo)的J774A.1遷移功能是否依賴HuR;胞質(zhì)蛋白的分離用于探究激活CB1是否能引起J774A.1胞質(zhì)中HuR的富集;RT-qPCR和Western blotting法檢測CB1和HuR mRNA和蛋白質(zhì)的變化情況。結(jié)果該研究證明J774A.1在基因和蛋白質(zhì)水平上均表達(dá)CB1和HuR;激活CB1能夠促進J774A.1的遷移(P0.01)并且能夠被其藥理學(xué)拮抗劑AM281所抑制;激活CB1誘導(dǎo)的J774A.1的遷移依賴HuR;激活CB1促進了J774A.1胞質(zhì)中HuR的富集進一步影響了CB1的表達(dá),由此HuR參與了激活CB1誘導(dǎo)的J774A.1的遷移。結(jié)論激活CB1能夠誘導(dǎo)單核巨噬細(xì)胞系J774A.1的遷移,且此過程依賴RNA結(jié)合蛋白HuR。
[Abstract]:Objective to study the important role of cannabinoid receptor 1(cannabinoid receptor 1 (CB1) in the migration of mononuclear macrophages and the possible mechanism of the involvement of RNA binding protein human antigen R(human antigen Ru Hur in the migration of mononuclear macrophages. Methods Mononuclear macrophage cell line J774A.1was used to identify the expression of CB1 and HuR in J774A.1 by agarose gel electrophoresis and immunofluorescence staining. ACEA and AM281 were pharmacological agonists and antagonists of CB1, respectively. The effect of ACEA and AM281 on J774A.1 migration activity was detected by Boyden chamber assay. The gene interference of Hur was used to determine whether the J774A.1 migration function induced by CB1 was dependent on HuR.The separation of cytoplasmic proteins was used to investigate whether activating CB1 could induce HuR in J774A.1 cytoplasm. The changes of CB1, HuR mRNA and protein were detected by RT-qPCR and Western blotting. Results this study demonstrated that J774A.1 expressed CB1 and Hur at both gene and protein levels, activation of CB1 promoted the migration of J774A.1 (P0.01) and was inhibited by its pharmacological antagonist AM281. Activation of CB1 induced migration of J774A.1 was dependent on Hur, and activation of CB1 promoted the accumulation of HuR in the cytoplasm of J774A.1 and further affected the expression of CB1, thus HuR was involved in CB1 induced J774A.1 migration. Conclusion Activation of CB1 can induce the migration of monocyte macrophage J774A.1, and this process is dependent on the RNA binding protein Hur.
【作者單位】： 首都醫(yī)科大學(xué)基礎(chǔ)醫(yī)學(xué)院細(xì)胞生物學(xué)系‘肝臟保護與再生調(diào)節(jié)’北京市重點實驗室;
【基金】：國家自然科學(xué)基金(81170407,31301154) 北京市屬高等學(xué)校創(chuàng)新團隊建設(shè)與教師職業(yè)發(fā)展計劃項目(IDHT20150502)~~
【分類號】：R392

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