Down regulation of UCP2 expression in retinal pigment epithelium cells under oxidative stress: an in vitro study
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Yuan He. Department of Ophthalmology, the Second Affiliated Hospital of Xi’an Medical University, Xi’an 710038, Shaanxi Province, China. openji7127@hotmail.com

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Supported by the National Natural Science Foundation of China (No.81100665; No.81770929).

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    Abstract:

    AIM: To evaluate the expression of uncoupling protein 2 (UCP2) in a retinal pigment epithelium cell line (ARPE-19), under oxidative stress (OS). METHODS: ARPE-19 cells were divided into groups treated with various concentrations of hydrogen peroxide (H2O2; 0, 150, 300, 500, 700, and 900 μmol/L) for 24h, to induce oxidative damage and cell viability was assessed by MTT assay. UCP2 mRNA expression in cells treated with H2O2 was investigated by reverse transcription-polymerase chain reaction (RT-PCR). UCP2 protein expression was assessed by Western blotting and ROS levels analyzed by flow cytometry (FCM). Further, UCP2-siRNA treated cultures were exposed to H2O2 (0, 75, 150, and 300 μmol/L) for 2h and cell viability determined by MTT assay. RESULTS: Cells treated with higher concentrations of H2O2 appeared shrunken; their adhesion to adjacent cells was disrupted, and the number of dead cells increased. The results of cell viability assays demonstrated that the numbers of cells were decreased in a dose-dependent manner following treatment with H2O2. Compared with untreated controls, cell viability was significantly reduced after treatment with >300 μmol/L H2O2 (P<0.05). Cell metabolic activity was decreased with increased concentrations of H2O2 as detected by MTT assay. Levels of OS were further decreased in cells treated with UCP2-siRNA compared with those treated with H2O2 alone (P<0.05). The results of RT-PCR and Western blotting demonstrated that UCP2 expression was reduced in H2O2-treated groups compared with controls (P<0.05). FCM analysis showed that cell reactive oxygen species (ROS) levels were increased in H2O2-treated groups and further upregulated by UCP2-siRNA treatment (P<0.05). CONCLUSION: Expression levels of UCP2 are decreased in ARPE-19 cells treated with H2O2. ROS levels are further increased in cells treated with UCP2-siRNA relative to those treated with H2O2 alone. UCP2 may have a protective role in ARPE-19 cells during oxidative injury.

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Ying Liu, Yuan Ren, Xia Wang, et al. Down regulation of UCP2 expression in retinal pigment epithelium cells under oxidative stress: an in vitro study. Int J Ophthalmol, 2019,12(7):1089-1094

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Publication History
  • Received:February 27,2018
  • Revised:May 06,2019
  • Adopted:
  • Online: June 11,2019
  • Published: