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BMC Ophthalmology
Published in: BMC Ophthalmology 1/2018

Open Access 01-12-2018 | Research article

UVB-mediated down-regulation of proteasome in cultured human primary pterygium fibroblasts

Authors: Alexios J. Aletras, Ioannis Trilivas, Maria-Elpida Christopoulou, Sotiria Drakouli, Constantine D. Georgakopoulos, Nikolaos Pharmakakis

Published in: BMC Ophthalmology | Issue 1/2018

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Abstract

Background

Pterygium is a condition characterized by epithelial overgrowth of the cornea, inflammatory cell infiltration and an abnormal extracellular matrix accumulation. Chronic UV exposure is considered as a pathogenic factor of this disease. Proteasome is an intracellular multi-subunit protease complex that degrades intracellular proteins. Among proteasome subunits the β5 (PSMB5), bearing chymotrypsin-like activity. It is considered as the main proteasome subunit and its expression is mediated by Nrf2-ARE pathway in many cell types. This study investigates the expression of PSMB5 in pterygium and the effect of UVB irradiation on its expression and activity in pterygium fibroblasts.

Methods

Normal conjunctival and pterygium specimens were obtained from the bulbar conjunctiva of patients undergoing cataract surgery and from patients with pterygium undergoing surgical removal of primary tissue, respectively. Fibroblasts were isolated upon treatment of specimens with clostridium collagenase. The expression of PSMB5 and Nrf2 in tissues and cells was ascertained by RT-PCR analysis and western blotting. Cell survival was measured by the MTT method and the proteasome chymotrypsin-like activity was determined by fluorometry.

Results

RT-PCR analysis showed that the expression of PSMB5 was significantly lower in pterygium than in normal conjunctiva. The expression of PSMB5 was mediated by the Nrf2/ARE pathway as indicated by using the Nrf2 activator Oltipraz. The expression of PSMB5 and Nrf2 by pterygium fibroblasts was suppressed in a dose dependent manner following UVB radiation of 0–50 mJ/cm2 doses. The expression of PSMB5, but not of Nrf2, remained at almost the control levels, when UVB exposure was performed after pre-incubation of cells with the src kinases inhibitor PP2. UVB irradiation had very low deleterious effect on fibroblasts survival, while it did not affect the proteasome chymotrypsin-like activity.

Conclusion

In pterygium fibroblasts, UVB exposure leads to down-regulation of Nrf2/ARE-mediated PSMB5 gene expression, in which src kinases may be implicated. This effect may be partially responsible for the lower expression of PSMB5 detected in pterygium as compared to normal conjunctiva.
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Metadata
Title
UVB-mediated down-regulation of proteasome in cultured human primary pterygium fibroblasts
Authors
Alexios J. Aletras
Ioannis Trilivas
Maria-Elpida Christopoulou
Sotiria Drakouli
Constantine D. Georgakopoulos
Nikolaos Pharmakakis
Publication date
01-12-2018
Publisher
BioMed Central
Published in
BMC Ophthalmology / Issue 1/2018
Electronic ISSN: 1471-2415
DOI
https://doi.org/10.1186/s12886-018-0987-8

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