腺苷受体在原代培养人视网膜色素上皮细胞中的表达研究

发布时间：2018-03-22 13:03

本文选题：视网膜色素上皮细胞　切入点：原代培养　出处：《重庆医科大学学报》2017年12期 　论文类型：期刊论文

【摘要】：目的:验证腺苷受体(adenosine receptors,ADORs)在原代培养的人视网膜色素上皮细胞(retinal pigment epithelium cells,RPE)的表达及其分布。方法:体外原代培养人RPE细胞。通过逆转录多聚酶链反应(transcription polymerase chain reaction,RT-PCR)检测RPE细胞中ADORs信使核糖核酸(messenger ribonucleic acid,mRNA)的表达情况,通过共聚焦免疫荧光法(confocal fluorescence microscopy,CFM)验证ADORs蛋白的表达并检测各亚型在RPE细胞内的分布。结果:在原代培养的RPE细胞中,ADORs亚型A1(adenosine receptors A1,ADORA1),ADORs亚型A2a(adenosine receptors A2a,ADORA2a)和ADORs亚型A2b(adenosine receptors A2b,ADORA2b)的mRNA及蛋白均有表达。ADORA1在RPE细胞的细胞核、核周及细胞质内均有表达,ADORA2a集中表达于RPE细胞核周及细胞质,ADORA2b在细胞核、核周及细胞质中强表达。ADORs亚型A3(adenosine receptors A3,ADORA3)在原代培养的RPE细胞中无表达。结论:原代培养RPE中均有腺苷受体ADORA1、ADORA2a、ADORA2b表达。腺苷受体各亚型在细胞内的不同表达位置可能提示了腺苷受体各亚型的不同功能。
[Abstract]:Objective: to investigate the expression and distribution of adenosine receptor adenosine receptor (ADORs) in primary cultured human retinal pigment epithelial cells (pigment epithelium cells). Methods: human RPE cells were cultured in vitro. Reverse transcriptase polymerase chain reaction (RT PCR) was used to detect the expression and distribution of adenosine receptor polymerase chain reactionsrRNA (RT-PCRs) in human retinal pigment epithelial cells (RPE). Methods: human RPE cells were cultured in vitro and detected by reverse transcription polymerase chain reaction (RT PCR). The expression of ADORs messenger ribonucleic aciddor mRNAs in RPE cells, Confocal fluorescence microscopy (CFM) was used to verify the expression of ADORs protein and to detect the distribution of ADORs subtypes in RPE cells. Results: the mRNA and ADORs subtypes A2b(adenosine receptors A2bORA2b of A1(adenosine receptors A1ADORA1 and A2a(adenosine receptors A2a ADORA2a) and ADORs subtype A2b(adenosine receptors A2bORA2b of primary cultured RPE cells were detected by confocal immunofluorescence. All the proteins expressed. ADORA1 in the nucleus of RPE cells. The expression of ADORA2a in the perinuclear and cytoplasm of RPE was concentrated in the periphery of the nucleus and the cytoplasm of ADORA2b in the nucleus. The strong expression of ADORs subtype A3(adenosine receptors A3A3ADORA3) was not found in primary cultured RPE cells. Conclusion: adenosine receptor ADORA1, ADORA2aORA2b and adenosine receptor subtypes may be expressed at different sites in primary cultured RPE. The different functions of adenosine receptor subtypes were demonstrated.
【作者单位】：重庆医科大学附属第一医院眼科眼科学重庆市市级重点实验室重庆市眼科研究所;眼科学国家重点实验室中山大学中山眼科中心;
【基金】：国家自然科学基金青年基金资助项目(编号:81300794)
【分类号】：R774.1

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