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  本文關(guān)鍵詞：缺氧對人RPE細(xì)胞表達(dá)HIF-1α的影響及T7-siRNAs對VEGF的干涉作用　出處：《吉林大學(xué)》2005年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 視網(wǎng)膜色素上皮細(xì)胞 HIF-1α RNA 干涉 脈絡(luò)膜新生血管

【摘要】：膜新生血管(choroidal neovascularization,CNV)可引起黃斑出血、滲出,最終瘢痕形成而導(dǎo)致患者視力喪失。視網(wǎng)膜色素上皮(retinal pigment epithelium,RPE)在CNV的形成過程中具有重要地位。我們首先利用胰蛋白酶消化的方法分離并原代培養(yǎng)了人視網(wǎng)膜色素上皮細(xì)胞,同時進(jìn)行了細(xì)胞鑒定。建立了hRPE細(xì)胞缺氧模型,并利用RT-PCR方法和免疫熒光方法檢測了不同缺氧時相hRPE細(xì)胞中HIF一1a轉(zhuǎn)錄和表達(dá)情況。RNA干涉作用(RNAi)是新發(fā)現(xiàn)的可以高效特異抑制基因表達(dá)途徑。我們根據(jù)VEGFl-5外顯子序列確定了3處靶位點(diǎn),并通過T7RNA聚合酶體外轉(zhuǎn)錄合成3對siRNA,轉(zhuǎn)染RPE細(xì)胞后,高效特異地干涉了hRPE細(xì)胞中VEGF的表達(dá)。這些實驗為研究CNV發(fā)生病理機(jī)制和基因治療CNV奠定了基礎(chǔ)。
[Abstract]:Choroidal neovascularization (CNV) may cause macular hemorrhage and exudation. Eventually scar formation results in visual loss. Retinal pigment epithelium (RPE) retinal pigment epithelium. RPE) plays an important role in the formation of CNV. Firstly, human retinal pigment epithelial cells were isolated and cultured by trypsin digestion. The anoxic model of hRPE cells was established. RT-PCR and immunofluorescence methods were used to detect the transcription and expression of HIF 1a in hRPE cells under different hypoxia conditions. We have identified three target sites according to the exon sequence of VEGFl-5. Three pairs of siRNAs were synthesized by T7 RNA polymerase transcription in vitro and transfected into RPE cells. These experiments provide a basis for studying the pathogenetic mechanism of CNV and gene therapy CNV.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2005
【分類號】：R363

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 鄭健粱,金學(xué)民,鄭湖玲,林建賢,馮官光;人視網(wǎng)膜色素上皮細(xì)胞的培養(yǎng)及鑒定[J];中國實用眼科雜志;1999年07期

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本文編號：1355676