1. Effect of CHI3L1 on the oxidative stress of human umbilical vein endothelial cells treated with palmitic acid
- Author
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Qiao Wang, Shu-Yue Zou, Shuai Chang, and Jia Zhou
- Subjects
human umbilical vein endothelial cells ,lcsh:R5-920 ,lcsh:R ,palmitic acid ,oxidative stress ,lcsh:Medicine ,chitinase 3-like protein 1 ,lcsh:Medicine (General) - Abstract
Objective To observe the effect of chitinase-3-like protein 1 (CHI3L1) on the oxidative stress of human umbilical vein endothelial cells (HUVECs) after treatment with palmitic acid, and preliminarily explore the molecular mechanism. Methods HUVECs were cultured in vitro, treated with different concentrations of CHI3L1 for 24 hours, and the MTT assay was used to determine the proliferation of HUVECs. The cells were divided into the control group (untreated group), palmitic acid group and CHI3L1+palmitic acid group. Cells in palmitic acid group were treated with 100 μmol/L palmitic acid for 24 hours, and in CHI3L1+palmitic acid group were pretreated with 100 ng/ml CHI3L1 for 2 hours, then 100 μmol/L palmitic acid were added and co-treating for 24 hours. Western blotting was used to detect the contents of nuclear and cytoplasmic p65, nuclear Lamin B, Actin, heme oxygenase (HMOX), reduced coenzyme Ⅰ/Ⅱ dependent quinone oxidoreductase (NQO1), phosphorylated protein kinase B (p-Akt), endoplasmic reticulum redox 1-like protein (Ero1-Lα), Calnexin, protein disulfide isomerase (PDI), chaperonin/ glucose regulatory protein (Bip1/Grp) 78, endoplasmic reticulum nuclear signal transduction protein (IRE)-1α and phosphorylated eukaryotic translation initiation factor (p-eIF) 2α. The contents of IL-6 and TNF-α were measured by ELISA, the activity of NF-κB was measured by DNA binding assay, and the nuclear translocation of Nrf2 was detected by immunofluorescence. Results MTT results showed that compared with the control, treatment with 100 ng/ml CHI3L1 for 24 hours did not decrease the activity of HUVECs (94.17±6.13 vs. 100.00±0.00). Western blotting results showed that the p65 level in nucleus of CHI3L1+palmitic acid group was lower than that in palmitic acid group (0.77±0.04 vs. 0.92±0.09, P
- Published
- 2020