Role of β-adrenergic receptor regulation of TNF-α and insulin signaling in retinal Müller cells

Robert J. Walker, Nancy M. Anderson, Youde Jiang, Suleiman Bahouth, Jena J. Steinle

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Purpose. The goal of this study was to determine the relationship of TNF-α and the downregulation of insulin receptor signaling in retinal Müller cells cultured under hyperglycemic conditions and the role of β-adrenergic receptors in regulating these responses. Methods. Retinal Müller cells were cultured in normal (5 mM) or high (25 mM) glucose until 80% confluent and then were reduced to 2% serum for 18 to 24 hours. The cells were then treated with 10 μM salmeterol followed by Western blot analysis or ELISA. For TNF-α inhibitory studies, the cells were treated with 5 ng/mL of TNF-α for 30 minutes or by a 30-minute pretreatment with TNF-α followed by salmeterol for 6 hours. In the TNF-α short hairpin (sh)RNA experiments, the cells were cultured until 90% confluent, followed by transfection with TNF-α shRNA for 18 hours. Results. TNF-α-only treatments of Müller cells resulted in significant decreases of tyrosine phosphorylation of the insulin receptor and Akt in high-glucose conditions. Salmeterol (10 μM), a β-2-adrenergic receptor agonist, significantly increased phosphorylation of both insulin receptor and Akt. TNF-α shRNA significantly decreased phosphorylation of IRS-1 Ser307, which was further decreased after salmeterol+TNF-α shRNA. Both TNF-α shRNA and salmeterol significantly reduced death of the retinal Müller cells. Conclusions. These studies demonstrate that β-adrenergic receptor agonists in vitro can restore the loss of insulin receptor activity noted in diabetes. By decreasing the levels of TNF-α and decreasing the phosphorylation of IRS-1 Ser307 while increasing tyrosine phosphorylation of insulin receptor, these results suggest a possible mechanism by which restoration of β-adrenergic receptor signaling may protect the retina against diabetes-induced damage.

Original languageEnglish (US)
Pages (from-to)9527-9533
Number of pages7
JournalInvestigative Ophthalmology and Visual Science
Volume52
Issue number13
DOIs
StatePublished - Dec 1 2011

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Insulin Receptor
Adrenergic Receptors
Small Interfering RNA
Phosphorylation
Insulin
Cultured Cells
Adrenergic Agonists
Tyrosine
Glucose
Transfection
Retina
Down-Regulation
Western Blotting
Enzyme-Linked Immunosorbent Assay
Salmeterol Xinafoate
Serum

All Science Journal Classification (ASJC) codes

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience
  • Medicine(all)

Cite this

Role of β-adrenergic receptor regulation of TNF-α and insulin signaling in retinal Müller cells. / Walker, Robert J.; Anderson, Nancy M.; Jiang, Youde; Bahouth, Suleiman; Steinle, Jena J.

In: Investigative Ophthalmology and Visual Science, Vol. 52, No. 13, 01.12.2011, p. 9527-9533.

Research output: Contribution to journalArticle

Walker, Robert J. ; Anderson, Nancy M. ; Jiang, Youde ; Bahouth, Suleiman ; Steinle, Jena J. / Role of β-adrenergic receptor regulation of TNF-α and insulin signaling in retinal Müller cells. In: Investigative Ophthalmology and Visual Science. 2011 ; Vol. 52, No. 13. pp. 9527-9533.
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abstract = "Purpose. The goal of this study was to determine the relationship of TNF-α and the downregulation of insulin receptor signaling in retinal M{\"u}ller cells cultured under hyperglycemic conditions and the role of β-adrenergic receptors in regulating these responses. Methods. Retinal M{\"u}ller cells were cultured in normal (5 mM) or high (25 mM) glucose until 80{\%} confluent and then were reduced to 2{\%} serum for 18 to 24 hours. The cells were then treated with 10 μM salmeterol followed by Western blot analysis or ELISA. For TNF-α inhibitory studies, the cells were treated with 5 ng/mL of TNF-α for 30 minutes or by a 30-minute pretreatment with TNF-α followed by salmeterol for 6 hours. In the TNF-α short hairpin (sh)RNA experiments, the cells were cultured until 90{\%} confluent, followed by transfection with TNF-α shRNA for 18 hours. Results. TNF-α-only treatments of M{\"u}ller cells resulted in significant decreases of tyrosine phosphorylation of the insulin receptor and Akt in high-glucose conditions. Salmeterol (10 μM), a β-2-adrenergic receptor agonist, significantly increased phosphorylation of both insulin receptor and Akt. TNF-α shRNA significantly decreased phosphorylation of IRS-1 Ser307, which was further decreased after salmeterol+TNF-α shRNA. Both TNF-α shRNA and salmeterol significantly reduced death of the retinal M{\"u}ller cells. Conclusions. These studies demonstrate that β-adrenergic receptor agonists in vitro can restore the loss of insulin receptor activity noted in diabetes. By decreasing the levels of TNF-α and decreasing the phosphorylation of IRS-1 Ser307 while increasing tyrosine phosphorylation of insulin receptor, these results suggest a possible mechanism by which restoration of β-adrenergic receptor signaling may protect the retina against diabetes-induced damage.",
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