RNA-Seq analysis of gene expression profiling in retinal vascular endothelial cells under high glucose condition_Chinese Journal of Ocular Fundus Diseases

Authors：

Zhang Zhe , Liu Juping , Dong Lijie , Liu Zhuqing , Huang Liangyu , Su Ruihong , Zhao Jinzhi , Zhang Xiaomin ,  Li Xiaorong

Tianjin Medical University Eye Hospital, Tianjin Medical University Eye Institute, Tianjin Medical University School of Optometry and Ophthalmology, Tianjin 300384, China;

Corresponding author：

Li Xiaorong, Email: iiitc1989@163.com

Keywords：

Retinal Vessels/cytology; Endothelial cells; Gene expression profiling; Diabetic retinopathy/etiology

DOI：

10.3760/cma.j.issn.1005-1015.2018.04.014

Video：

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ObjectiveTo observe RNA-Seq analysis of gene expression profiling in retinal vascular endothelial cells after anti-vascular endothecial growth factor (VEGF) treatment.MethodsRetinal vascular endothelial cells were cultured in vitro, and the logarithmic growth phase cells were used for experiments. The cells were divided into the control group and high glucose group. The cells of two groups were cultured for 5 hours with 5, 25 mmol/L glucose, respectively. And then, whole transcriptome sequencing approach was applied to the above two groups of cells through RNA-Seq. Now with biological big data obtained as a basis, to analyze the differentially expressed genes (DEGs). And through enrichment analysis to explain the differential functions of DEGs and their signal pathways.ResultsThe gene expression profiles of the two groups of cells were obtained. Through analysis, 449 DEGs were found, including 297 upregulated and 152 downregulated ones. The functions of DEGs were influenced by regulations over molecular biological process, cellular energy metabolism and protein synthesis, etc. Among these genes, ITGB1BP2, NCF1 and UNC5C were related to production of inflammation; AKR1C4, ATP1A3, CHST5, LCTL were related to energy metabolism of cells; DAB1 and PRSS55 were related to protein synthesis; SMAD9 and BMP4 were related to the metabolism of extracellular matrix. GO enrichment analysis showed that DEGs mainly act in three ways: regulating biological behavior, organizing cellular component and performing molecular function, which were mainly concentrated in the system generation of biological process part and regulation of multicellular organisms. Pathway enrichment analysis showed that gene expressions of the two cell groups were differentiated in transforming growth factor-β (TGF-β) signaling pathway, complement pathway and amino acid metabolism-related pathways have also been affected, such as tryptophan, serine and cyanide. Among them, leukocyte inhibitory factor 9 and bone morphogenetic protein 4 play a role through the TGF-β signaling pathway.ConclusionsHigh glucose affects the function of retinal vascular endothelial cells by destroying transmembrane conduction of retinal vascular endothelial cells, metabolism of extracellular matrix, and transcription and translation of proteins.

Citation： Zhang Zhe, Liu Juping, Dong Lijie, Liu Zhuqing, Huang Liangyu, Su Ruihong, Zhao Jinzhi, Zhang Xiaomin, Li Xiaorong. RNA-Seq analysis of gene expression profiling in retinal vascular endothelial cells under high glucose condition. Chinese Journal of Ocular Fundus Diseases, 2018, 34(4): 377-381. doi: 10.3760/cma.j.issn.1005-1015.2018.04.014 Copy

1.	Kawano H, Motoyama T, Hirashima O, et al. Hyperglycemia rapidly suppresses flow-mediated endothelium- dependent vasodilation of brachial artery[J]. J Am Coll Cardiol, 1999, 34(1):146-154.
2.	Balletshofer BM, Rittig K, Enderle MD, et al. Endothelial dysfunction is detectable in young normotensive first-degree relatives of subjects with type 2 diabetes in association with insulin resistance[J]. Circulation, 2000, 101(15):1780-1784.
3.	Wang Z, Gerstein M, Snyder M. RNA-Seq: a revolutionary tool for transcriptomics[J]. Nat Rev Genet, 2009, 10(1):57-63. DOI: 10.1038/nrg2484.
4.	Mutz KO, Heilkenbrinker A, Lönne M, et al. Transcriptome analysis using next-generation sequencing[J]. Curr Opin Biotechnol, 2013, 24(1):22-30. DOI: 10.1016/j.copbio.2012.09.004.
5.	Mantione KJ, Kream RM, Kuzelova H, et al. Comparing bioinformatic gene expression profiling methods: microarray and RNA-Seq[J]. Med Sci Monit Basic Res, 2014, 20:138-142. DOI: 10.12659/MSMBR.892101.
6.	牛瑞, 东莉洁, 胡博杰, 等. 抗血管内皮生长因子药物治疗后视网膜血管内皮细胞基因表达谱的RNA—Seq分析[J].中华眼底病杂志,2018,34(3):275-280. DOI: 10.3760/cma.j.issn.1005-1015.2018.03.016.Niu R, Dong LJ, Hu BJ, et al. RNA-Seq analysis of gene expression profiling in human retinal vascular endothelial cells after anti-vascular endothecial growth factor treatment [J]. Chin J Ocul Fundus Dis, 2018, 34(3):275-280. DOI: 10.3760/cma.j.issn.1005-1015.2018.03.016.
7.	Derynck R, Zhang YE. Smad-dependent and Smad-independent pathways in TGF-beta family signalling[J]. Nature, 2003, 425(6958):577-584. DOI: 10.1038/nature02006.
8.	Attisano L, Wrana JL. Signal transduction by the TGF-beta superfamily[J]. Science, 2002, 296(5573):1646-1647. DOI: 10.1126/science.1071809.
9.	Wan TT, Li XF, Sun YM, et al. Recent advances in understanding the biochemical and molecular mechanism of diabetic retinopathy[J]. Biomed Pharmacother,2015,74:145-147. DOI: 10.1016/j.biopha.2015.08.002.
10.	Lee S, Clark SA, Gill RK, et al.1,25-dihydroxyvitamin D3 and pancreatic β-cell function: vitamin D receptors, gene expression and insulin secretion[J]. Endocrinology,1994,134(4):1602-1610. DOI: 10.1210/endo.134.4.8137721.
11.	Carroll MC. The complement system in regulation of adaptive immunity[J]. Nature Immunology, 2004, 5(10):981-986.
12.	Uesugi N, Sakata N, Nangaku M, et al. Possible mechanism for medial smooth muscle cell injury in diabetic nephropathy: glycoxidation-mediated local complement activation[J]. Am J Kidney Dis, 2004, 44(2):224-238.
13.	Zhang J, Gerhardinger CM. Early complement activation and decreased levels of glycosylphosphatidylinositol-anchored complement inhibitors in human and experimental diabetic retinopathy[J]. Diabetes, 2002, 51(12):3499-3504.
14.	Qin X, Goldfine A, Krumrei N, et al. Glycation inactivation of the complement regulatory protein CD59: a possible role in the pathogenesis of the vascular complications of human[J]. Diabetes, 2004, 53(10):2653-2661.
15.	Gerl VB, Bohl J, Pitz S, et al. Extensive deposits of complement C3d and C5b-9 in the choriocapillaris of eyes of patients with diabetic retinopathy[J]. Invest Ophthalmol Vis Sci, 2002, 43(4):1104-1108.
16.	Peng Q, Li K, Smyth LA, et al. C3a and C5a promote renal ischemia-reperfusion injury[J]. J Am Soc Nephrol, 2012, 23(9):1474-1485. DOI: 10.1681/ASN.2011111072.
17.	Peng Q, Li K, Sacks S H, et al. The role of anaphylatoxins C3a and C5a in regulating innate and adaptive immune responses[J]. Inflamm Allergy Drug Targets, 2009, 8(3):236-246.
18.	Segal BH, Grimm MJ, Khan AN, et al. Regulation of innate immunity by NADPH oxidase[J]. Free Radic Biol Med, 2012, 53(1):72-80. DOI: 10.1016/j.freeradbiomed.2012.04.022.
19.	Segal BH, Veys P, Malech H, et al. Chronic granulomatous disease: lessons from a rare disorder[J]. Biol Blood Marrow Transplant, 2011, 17(1):123-131. DOI: 10.1016/j.bbmt.2010.09.008.
20.	Adaikalakoteswari A, Balasubramanyam M, Rema M, et al. Differential gene expression of NADPH oxidase (p22phox) and hemoxygenase-1 in patients with Type 2 diabetes and microangiopathy[J]. Diabet Med, 2006, 23(6):666-674.

1. Kawano H, Motoyama T, Hirashima O, et al. Hyperglycemia rapidly suppresses flow-mediated endothelium- dependent vasodilation of brachial artery[J]. J Am Coll Cardiol, 1999, 34(1):146-154.
2. Balletshofer BM, Rittig K, Enderle MD, et al. Endothelial dysfunction is detectable in young normotensive first-degree relatives of subjects with type 2 diabetes in association with insulin resistance[J]. Circulation, 2000, 101(15):1780-1784.
3. Wang Z, Gerstein M, Snyder M. RNA-Seq: a revolutionary tool for transcriptomics[J]. Nat Rev Genet, 2009, 10(1):57-63. DOI: 10.1038/nrg2484.
4. Mutz KO, Heilkenbrinker A, Lönne M, et al. Transcriptome analysis using next-generation sequencing[J]. Curr Opin Biotechnol, 2013, 24(1):22-30. DOI: 10.1016/j.copbio.2012.09.004.
5. Mantione KJ, Kream RM, Kuzelova H, et al. Comparing bioinformatic gene expression profiling methods: microarray and RNA-Seq[J]. Med Sci Monit Basic Res, 2014, 20:138-142. DOI: 10.12659/MSMBR.892101.
6. 牛瑞, 东莉洁, 胡博杰, 等. 抗血管内皮生长因子药物治疗后视网膜血管内皮细胞基因表达谱的RNA—Seq分析[J].中华眼底病杂志,2018,34(3):275-280. DOI: 10.3760/cma.j.issn.1005-1015.2018.03.016.Niu R, Dong LJ, Hu BJ, et al. RNA-Seq analysis of gene expression profiling in human retinal vascular endothelial cells after anti-vascular endothecial growth factor treatment [J]. Chin J Ocul Fundus Dis, 2018, 34(3):275-280. DOI: 10.3760/cma.j.issn.1005-1015.2018.03.016.
7. Derynck R, Zhang YE. Smad-dependent and Smad-independent pathways in TGF-beta family signalling[J]. Nature, 2003, 425(6958):577-584. DOI: 10.1038/nature02006.
8. Attisano L, Wrana JL. Signal transduction by the TGF-beta superfamily[J]. Science, 2002, 296(5573):1646-1647. DOI: 10.1126/science.1071809.
9. Wan TT, Li XF, Sun YM, et al. Recent advances in understanding the biochemical and molecular mechanism of diabetic retinopathy[J]. Biomed Pharmacother,2015,74:145-147. DOI: 10.1016/j.biopha.2015.08.002.
10. Lee S, Clark SA, Gill RK, et al.1,25-dihydroxyvitamin D3 and pancreatic β-cell function: vitamin D receptors, gene expression and insulin secretion[J]. Endocrinology,1994,134(4):1602-1610. DOI: 10.1210/endo.134.4.8137721.
11. Carroll MC. The complement system in regulation of adaptive immunity[J]. Nature Immunology, 2004, 5(10):981-986.
12. Uesugi N, Sakata N, Nangaku M, et al. Possible mechanism for medial smooth muscle cell injury in diabetic nephropathy: glycoxidation-mediated local complement activation[J]. Am J Kidney Dis, 2004, 44(2):224-238.
13. Zhang J, Gerhardinger CM. Early complement activation and decreased levels of glycosylphosphatidylinositol-anchored complement inhibitors in human and experimental diabetic retinopathy[J]. Diabetes, 2002, 51(12):3499-3504.
14. Qin X, Goldfine A, Krumrei N, et al. Glycation inactivation of the complement regulatory protein CD59: a possible role in the pathogenesis of the vascular complications of human[J]. Diabetes, 2004, 53(10):2653-2661.
15. Gerl VB, Bohl J, Pitz S, et al. Extensive deposits of complement C3d and C5b-9 in the choriocapillaris of eyes of patients with diabetic retinopathy[J]. Invest Ophthalmol Vis Sci, 2002, 43(4):1104-1108.
16. Peng Q, Li K, Smyth LA, et al. C3a and C5a promote renal ischemia-reperfusion injury[J]. J Am Soc Nephrol, 2012, 23(9):1474-1485. DOI: 10.1681/ASN.2011111072.
17. Peng Q, Li K, Sacks S H, et al. The role of anaphylatoxins C3a and C5a in regulating innate and adaptive immune responses[J]. Inflamm Allergy Drug Targets, 2009, 8(3):236-246.
18. Segal BH, Grimm MJ, Khan AN, et al. Regulation of innate immunity by NADPH oxidase[J]. Free Radic Biol Med, 2012, 53(1):72-80. DOI: 10.1016/j.freeradbiomed.2012.04.022.
19. Segal BH, Veys P, Malech H, et al. Chronic granulomatous disease: lessons from a rare disorder[J]. Biol Blood Marrow Transplant, 2011, 17(1):123-131. DOI: 10.1016/j.bbmt.2010.09.008.
20. Adaikalakoteswari A, Balasubramanyam M, Rema M, et al. Differential gene expression of NADPH oxidase (p22phox) and hemoxygenase-1 in patients with Type 2 diabetes and microangiopathy[J]. Diabet Med, 2006, 23(6):666-674.

Chinese Journal of Ocular Fundus Diseases

RNA-Seq analysis of gene expression profiling in retinal vascular endothelial cells under high glucose condition

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