Thioredoxin induced antioxidant gene expressions in human lens epithelial cells.
Thioredoxin (Trx) is one of the major redox-regulating proteins. It catalyzes dithiol/disulfide exchange reactions and displays many unique intracellular and extracellular activities thereby controlling multiple mammalian cell functions. In the present study we examine the effect of exogenous Trx on the expression of several antioxidant genes in human lens epithelial (HLE B3) cells. mRNA levels for gene expression were monitored by RT-PCR and real-time PCR while protein levels were measured by western blot analysis. We have found that recombinant human Trx (hTrx)-treated HLE B3 cells have a simultaneous increase in mRNA expressions of mitochondrial manganese superoxide dismutase (MnSOD), thioltranferase 1 (TTase 1) or glutaredoxin 1 (Grx1), mitochondrial thioltransferase (TTase 2) or glutaredoxin 2 (Grx2), and thioredoxin peroxidase IV (Prx IV). The increased MnSOD and TTase 1 mRNA expressions were accompanied with their respective increases in protein levels. Other antioxidant genes, including Cu/ZnSOD, catalase, glutathione peroxidase 1 (GPx1), thioredoxin reductase 1 (TrxR1), thioredoxin peroxidase III (Prx III), and gamma-glutamyl cysteine synthetase were not affected. The ability of Trx to induce selectively these antioxidant genes in the absence of oxidative stress suggest a cytokine/growth factor-like new physiological role of hTrx in HLE B3 cells. Our data also provide evidence of a strong antioxidant defense system in HLE B3 cells that can be activated by extracellular hTrx, as well as of a possible link between the thioredoxin (Trx) and glutathione (GSH) redox regulating systems in these cells.
Yegorova
Svitlana
S
Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln, 134 VBS, Fair St./East Campus Loop, 68583-0905, USA.
Yegorov
Oleg
O
Lou
Marjorie F
MF
D000975:Antioxidants / Q000378:metabolism*; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000187:drug effects / Q000201:enzymology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D054477:Glutaredoxins; D006801:Humans; D007908:Lens, Crystalline / Q000187:drug effects* / Q000201:enzymology; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010088:Oxidoreductases / Q000096:biosynthesis / Q000235:genetics; D010544:Peroxidases / Q000096:biosynthesis / Q000235:genetics; D054464:Peroxiredoxins; D011490:Protein Disulfide Reductase (Glutathione) / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013482:Superoxide Dismutase / Q000096:biosynthesis / Q000235:genetics; D013879:Thioredoxins / Q000494:pharmacology*
D000975:Antioxidants; C516008:GLRX2 protein, human; D054477:Glutaredoxins; D011994:Recombinant Proteins; D013879:Thioredoxins; D010088:Oxidoreductases; D010544:Peroxidases; D054464:Peroxiredoxins; D013482:Superoxide Dismutase; D011490:Protein Disulfide Reductase (Glutathione)