Glutaredoxin regulates autocrine and paracrine proinflammatory responses in retinal glial (muller) cells.
Protein S-glutathionylation is a reversible redox-dependent post-translational modification. Many cellular functions and signal transduction pathways involve proteins whose cysteine-dependent activities are modulated by glutathionylation. Glutaredoxin (Grx1) plays a key role in such regulation because it is a specific and efficient catalyst of deglutathionylation. We recently reported an increase in Grx1 in retinae of diabetic rats and in rat retinal Müller glial cells (rMC-1) cultured in high glucose. This up-regulation of Grx1 was concomitant with NFkappaB activation and induction of intercellular adhesion molecule-1 (ICAM-1). This proinflammatory response was replicated by adenoviral-directed up-regulation of Grx1 in cells in normal glucose. The site of regulation of NFkappaB was localized to the cytoplasm, where IkappaB kinase (IKK) is a master regulator of NFkappaB activation. In the current study, inhibition of IKK activity abrogated the increase in ICAM-1 induced by high glucose or by adenoviral-directed up-regulation of Grx1. Conditioned medium from the Müller cells overexpressing Grx1 was added to fresh cultures of Müller or endothelial cells and elicited increases in the Grx1 and ICAM-1 proteins in these cells. These effects correlate with a novel finding that secretion of interleukin-6 was elevated in the cultures of Grx overexpressing cells. Also, pure interleukin-6 increased Grx1 and ICAM-1 in the rMC-1 cells. Thus, Grx1 appears to play an important role in both autocrine and paracrine proinflammatory responses. Furthermore, IKKbeta isolated from Müller cells in normal glucose medium was found to be glutathionylated on Cys-179. Hence Grx-mediated activation of IKK via deglutathionylation may play a central role in diabetic complications in vivo where Grx1 is increased.
Shelton
Melissa D
MD
Department of Pharmacology, Case Western Reserve University School of Medicine, Cleveland, OH 44106-4965, USA.
Distler
Anne M
AM
Kern
Timothy S
TS
Mieyal
John J
JJ
D000256:Adenoviridae; D000818:Animals; D019898:Autocrine Communication; D002461:Cell Line, Transformed; D048909:Diabetes Complications / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000235:genetics; D005947:Glucose / Q000378:metabolism; D054477:Glutaredoxins / Q000096:biosynthesis* / Q000235:genetics; D051550:I-kappa B Kinase / Q000235:genetics / Q000378:metabolism; D007249:Inflammation / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis / Q000235:genetics; D015850:Interleukin-6 / Q000096:biosynthesis / Q000494:pharmacology; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D009457:Neuroglia / Q000201:enzymology* / Q000473:pathology; D019899:Paracrine Communication; D051381:Rats; D012160:Retina / Q000201:enzymology* / Q000473:pathology; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics
D054477:Glutaredoxins; D015850:Interleukin-6; D016328:NF-kappa B; D018799:Intercellular Adhesion Molecule-1; D051550:I-kappa B Kinase; D005947:Glucose