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Human lens thioredoxin: molecular cloning and functional characterization
Svitlana Yegorova1, Aimin Liu, Marjorie F Lou
1Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, Nebraska 68583-0905, USA.
Investigative Ophthalmology & Visual Science
|July 29, 2003
Summary
The thioredoxin system, specifically thioredoxin 1 (Trx1), is upregulated in human lens epithelial cells under oxidative stress, suggesting its protective role in the lens.
Area of Science:
- Ophthalmology
- Molecular Biology
- Biochemistry
Background:
- The human lens requires protection against oxidative damage.
- Thioredoxin 1 (Trx1) is a key protein in cellular redox homeostasis.
Purpose of the Study:
- To clone the human lens thioredoxin (TXN) gene.
- To characterize recombinant Trx1 protein.
- To investigate Trx1 expression regulation in human lens epithelial cells under oxidative stress.
Main Methods:
- Human TXN gene cloning from a lens cDNA library.
- Insulin reduction assay for Trx1 activity.
- Exposure of human lens epithelial cells (HLE B3) to H(2)O(2) for oxidative stress.
- Western blot, real-time PCR, and Trx1 activity assays.
Main Results:
- Human lens TXN gene sequence is conserved.
- Recombinant Trx1 exhibits resistance to oxidation.
- Trx1 is expressed in HLE B3 cells and localized in human and porcine lenses.
- Oxidative stress upregulated Trx1 protein by 35% and mRNA by 50%.
Conclusions:
- Trx1 upregulation in response to oxidative stress indicates its role in lens defense.
- The thioredoxin system is a potential protective mechanism against oxidative damage in the lens.