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体外人视网膜色素上皮细胞的光凝:通过转录组学和蛋白质组学揭示对ARPE-19的影响

Photocoagulation of human retinal pigment epithelium in vitro: unravelling the effects on ARPE-19 by transcriptomics and proteomics.

作者信息

Tababat-Khani Poya, de la Torre Carolina, Canals Francesc, Bennet Hedvig, Simo Rafael, Hernandez Cristina, Fex Malin, Agardh Carl-David, Hansson Ola, Agardh Elisabet

机构信息

Unit on Vascular Diabetic Complications, Department of Clinical Sciences, Lund University Diabetes Center, Malmö, Sweden.

Proteomics Laboratory, Vall d'Hebron Institute of Oncology (VHIO), Vall d'Hebron University Hospital, Universitat Autònoma de Barcelona, Barcelona, Spain.

出版信息

Acta Ophthalmol. 2015 Jun;93(4):348-54. doi: 10.1111/aos.12649. Epub 2015 Jan 21.

Abstract

PURPOSE

Despite the extensive use of retinal photocoagulation for ischaemia and vascular leakage in retinal vascular disease, the molecular mechanisms behind its clinical beneficial effects are still poorly understood. One important target of laser irradiation is the retinal pigment epithelium (RPE). In this study, we aimed at identifying the isolated effects of photocoagulation of RPE at both the mRNA and protein expression levels.

METHODS

Human ARPE-19 cells were exposed to photocoagulation. Gene expression and protein expression were compared to untreated cells using microarray and liquid chromatography-mass spectrometry analysis. Genes and proteins queried by microarray and mass spectrometry were subjected to the Kyoto Encyclopedia of Genes and Genomes (KEGG) database pathway analyses.

RESULTS

Laser irradiation resulted in an induction of the cytoprotective heat-shock protein subfamily Hsp70 as well as in a suppression of the vascular permeability factor carbonic anhydrase 9 (CA9). These expression patterns were evident at both the mRNA and protein levels. KEGG pathway analyses revealed genes and proteins involved in cellular turnover, repair and inflammation.

CONCLUSIONS

By characterizing the transcriptional and translational effects of laser coagulation on the RPE cells in culture, we have revealed responses, which might contribute to some of the beneficial effects obtained by photocoagulation for ischaemia and vascular leakage in retinal vascular disease.

摘要

目的

尽管视网膜光凝术在视网膜血管疾病的缺血和血管渗漏治疗中被广泛应用,但其临床有益效果背后的分子机制仍知之甚少。激光照射的一个重要靶点是视网膜色素上皮(RPE)。在本研究中,我们旨在确定RPE光凝在mRNA和蛋白质表达水平上的单独作用。

方法

将人ARPE - 19细胞暴露于光凝环境中。使用微阵列和液相色谱 - 质谱分析将基因表达和蛋白质表达与未处理的细胞进行比较。对微阵列和质谱检测到的基因和蛋白质进行京都基因与基因组百科全书(KEGG)数据库通路分析。

结果

激光照射导致细胞保护性热休克蛋白亚家族Hsp70的诱导以及血管通透性因子碳酸酐酶9(CA9)的抑制。这些表达模式在mRNA和蛋白质水平均很明显。KEGG通路分析揭示了参与细胞更新、修复和炎症的基因和蛋白质。

结论

通过表征激光凝固对培养的RPE细胞的转录和翻译作用,我们揭示了一些反应,这些反应可能有助于解释视网膜血管疾病中光凝术在缺血和血管渗漏方面所获得的一些有益效果。

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