本文選題：視網(wǎng)膜色素上皮細胞　切入點：原代培養(yǎng)　出處：《重慶醫(yī)科大學(xué)學(xué)報》2017年12期 　論文類型：期刊論文

【摘要】：目的:驗證腺苷受體(adenosine receptors,ADORs)在原代培養(yǎng)的人視網(wǎng)膜色素上皮細胞(retinal pigment epithelium cells,RPE)的表達及其分布。方法:體外原代培養(yǎng)人RPE細胞。通過逆轉(zhuǎn)錄多聚酶鏈反應(yīng)(transcription polymerase chain reaction,RT-PCR)檢測RPE細胞中ADORs信使核糖核酸(messenger ribonucleic acid,mRNA)的表達情況,通過共聚焦免疫熒光法(confocal fluorescence microscopy,CFM)驗證ADORs蛋白的表達并檢測各亞型在RPE細胞內(nèi)的分布。結(jié)果:在原代培養(yǎng)的RPE細胞中,ADORs亞型A1(adenosine receptors A1,ADORA1),ADORs亞型A2a(adenosine receptors A2a,ADORA2a)和ADORs亞型A2b(adenosine receptors A2b,ADORA2b)的mRNA及蛋白均有表達。ADORA1在RPE細胞的細胞核、核周及細胞質(zhì)內(nèi)均有表達,ADORA2a集中表達于RPE細胞核周及細胞質(zhì),ADORA2b在細胞核、核周及細胞質(zhì)中強表達。ADORs亞型A3(adenosine receptors A3,ADORA3)在原代培養(yǎng)的RPE細胞中無表達。結(jié)論:原代培養(yǎng)RPE中均有腺苷受體ADORA1、ADORA2a、ADORA2b表達。腺苷受體各亞型在細胞內(nèi)的不同表達位置可能提示了腺苷受體各亞型的不同功能。
[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.
【作者單位】： 重慶醫(yī)科大學(xué)附屬第一醫(yī)院眼科眼科學(xué)重慶市市級重點實驗室重慶市眼科研究所;眼科學(xué)國家重點實驗室中山大學(xué)中山眼科中心;
【基金】：國家自然科學(xué)基金青年基金資助項目(編號:81300794)
【分類號】：R774.1

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