Quantitative proteomic analysis of human retinal microvascular endothelial cells stimulated with 4-Hydroxynonenal
10.3760/cma.j.issn.1005-1015.2019.05.012
- VernacularTitle:4-羟基壬稀酸刺激人视网膜微血管内皮细胞的定量蛋白质组学研究
- Author:
Jing XIAO
1
;
Xiaomin ZHANG
;
Xianfeng SHAO
;
Mengran XIAO
;
Fuhua YANG
;
Hui ZHANG
;
L Vicki EA
;
Xiaorong LI
Author Information
1. 天津医科大学眼科医院、眼视光学院、眼科研究所天津市视网膜功能与疾病重点实验室 300384
- Keywords:
Reitinal vessels/cytology;
Endothelial;
Cell cultured;
Proteomics;
4- Hydroxynonenal
- From:
Chinese Journal of Ocular Fundus Diseases
2019;35(5):488-493
- CountryChina
- Language:Chinese
-
Abstract:
Objective To detect the protein expression change in the proliferation of human retinal microvascular endothelial cells (hRMECs) stimulated with 4-Hydroxynonenal (4-HNE).Methods hRMECs were in a logarithmic growth phase, and then were separated into 4-HNE-stimulated group and negative control group. The concentration of 4-HNE included 5, 10, 20 and 50 μmol/L in 4-HNE-stimulated group, while the negative control group was added in the same volume of ethanol (the solvent of 4-HNE). Then the cells were stimulated with 4-HNE for 24 hours following by CCK-8 kits incubating for 4 hours to detect absorbance. It was found that 10 μmol/L 4-HNE had the most obvious effect on the proliferation of hRMECs. Therefore, the cellular proteins from 10 μmol/L 4-HNE-stimulated group and negative control group were acquired and prepared by FASP sample preparation method. Data independent acquisition was used for data acquisition, and the GO analysis and pathway enrichment were performed for analysis of differentially expressed proteins. Results CCK-8 kits detection results showed that theA value of the 10 and 20 μmol/L 4-HNE-stimulated groups were significantly higher than negative control group and 5 μmol/L 4-HNE-stimulated group (F=25.42, P<0.01), while there were no differences between 10 and 20 μmol/L 4-HNE-stimulated groups, and theA value of 50 μmol/L 4-HNE-stimulated groups was lower than negative control. A total of 2710 quantifiable proteins were identified by peoteomics,and 118 proteins were differentially expressed (fold change>1.5,P<0.05). Seventy-two proteins were up-regulated after 4-HNE stimulation, whereas 46 proteins were down-regulated. Particularly, the expressions of Heme oxygenase-1, Sulfoxdoxin-1, Heat shock protein A1B, Thioredoxin reductase-1, Glutathione reductase, ATPase and prothrombin were increased when cells were added in 4-HNE, whereas the expressions of apolipoprotein A1 and programmed cell death protein 4 were decreased. These differentially expressed proteins were mainly involved in the biological processes such as oxidative stress, cell detoxification, and ATPase-coupled membrane transport.Conclusion After stimulated with 4-HNE, the oxidative stress, cell detoxification, and ATPase-coupled membrane transport protein expression may change in hRMECs in order to regulate oxidative stress and growth situation.
