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

万文娟; 崔冬梅; 曾骏文; 李琦

doi:10.13406/j.cnki.cyxb.001356

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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

基础研究 | 更新时间：2024-01-17

- 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
- Expression of adenosine receptors in human retinal pigment epithelium cells primary cultured in vitro
- 重庆医科大学学报 2017年42卷第12期页码：1587-1591
- 作者机构：
  
  1. 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
  2. 眼科学国家重点实验室中山大学中山眼科中心
- 作者简介：
- 基金信息：
  
  国家自然科学基金青年基金资助项目(编号:81300794)
- DOI：10.13406/j.cnki.cyxb.001356
  中图分类号： R774.1
- 纸质出版日期：2017，
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万文娟, 崔冬梅, 曾骏文, 等. 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[J]. 重庆医科大学学报, 2017,42(12):1587-1591.

Wan Wenjuan, Cui Dongmei, Zeng Junwen, et al. Expression of adenosine receptors in human retinal pigment epithelium cells primary cultured in vitro[J]. Journal of Chongqing Medical University, 2017,42(12):1587-1591.
万文娟, 崔冬梅, 曾骏文, 等. 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[J]. 重庆医科大学学报, 2017,42(12):1587-1591. DOI： 10.13406/j.cnki.cyxb.001356.

Wan Wenjuan, Cui Dongmei, Zeng Junwen, et al. Expression of adenosine receptors in human retinal pigment epithelium cells primary cultured in vitro[J]. Journal of Chongqing Medical University, 2017,42(12):1587-1591. DOI： 10.13406/j.cnki.cyxb.001356.

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 determine the distribution of adenosine receptors（ADORs）in primary cultured human retinal pigment epithelium（RPE）cells.Methods:Human RPE cells were primary cultured in vitro.ADORs messenger ribonucleic acid（mRNA）in RPE was detected by reverse transcription polymerase chain reaction（RT-PCR）.Confocal fluorescence microscopy was used to confirm the protein expression and study the subcellular distribution of ADORs.Results:Adenosine receptors A1

A2a and A2 b（ADORA1

ADORA2a and ADORA2b）mRNA and protein were expressed in primary cultured RPE cells.The ADORs subtypes were differently distributed within the cells.ADORA1 was expressed in nucleus

perinucleus and cytoplasm of RPE.ADORA2a was concentrated mainly in perinucleus and cytoplasm.ADORA2b was strongly expressed in the nucleus

perinucleus and the cytoplasm.Adenosine receptors A3（ADORA3）was barely expressed in the primary cultured RPE cells.Conclusion:ADORA1

ADORA2a

ADORA2b are expressed in human RPE cells primary cultured in vitro.The different distributions at the subcellular level suggests different functions of ADOR subtypes.

关键词

视网膜色素上皮细胞原代培养腺苷受体

Keywords

human retinal pigment epithelium cellsprimary cultureadenosine receptors

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