Exp Mol Med.  2011 Dec;43(12):684-692. 10.3858/emm.2011.43.12.079.

Inhibition of NF-kappaB prevents high glucose-induced proliferation and plasminogen activator inhibitor-1 expression in vascular smooth muscle cells

• Jeong IK
• Oh DH
• Park SJ
• Kang JH
• Kim S
• Lee MS
• Kim MJ
• Hwang YC
• Ahn KJ
• Chung HY
• Chae MK
• Yoo HJ

• KMID: 1058706
• DOI: http://doi.org/10.3858/emm.2011.43.12.079

Abstract

Recent epidemiologic studies clearly showed that early intensive glucose control has a legacy effect for preventing diabetic macrovascular complications. However, the cellular and molecular processes by which high glucose leads to macrovascular complications are poorly understood. Vascular smooth muscle cell (VSMC) dysfunction due to high glucose is a characteristic of diabetic vascular complications. Activation of nuclear factor-kappaB (NF-kappaB) may play a key role in the regulation of inflammation and proliferation of VSMCs. We examined whether VSMC proliferation and plasminogen activator inhibitor-1 (PAI-1) expression induced by high glucose were mediated by NF-kappaB activation. Also, we determined whether selective inhibition of NF-kappaB would inhibit proliferation and PAI-1 expression in VSMCs. VSMCs of the aorta of male SD rats were treated with various concentrations of glucose (5.6, 11.1, 16.7, and 22.2 mM) with or without an inhibitor of NF-kappaB or expression of a recombinant adenovirus vector encoding an IkappaB-alpha mutant (Ad-IkappaBalphaM). VSMC proliferation was examined using an MTT assay. PAI-1 expression was assayed by real-time PCR and PAI-1 protein in the media was measured by ELISA. NF-kappaB activation was determined by immunohistochemical staining, NF-kappaB reporter assay, and immunoblotting. We found that glucose stimulated VSMC proliferation and PAI-1 expression in a dose-dependent manner up to 22.2 mM. High glucose (22.2 mM) alone induced an increase in NF-kappaB activity. Treatment with inhibitors of NF-kappaB such as MG132, PDTC or expression of Ad-IkappaB-alphaM in VSMCs prevented VSMC proliferation and PAI-1 expression induced by high glucose. In conclusion, inhibition of NF-kappaB activity prevented high glucose-induced VSMC proliferation and PAI-1 expression.

Keyword

cell proliferation; glucose metabolism disorders; muscle, smooth, vascular; NF-kappaB; plasminogen activator inhibitor 1

MeSH Terms

Animals
Aorta/cytology
Cardiovascular Diseases/prevention & control
Cell Proliferation/*drug effects
Cells, Cultured
Diabetes Complications/prevention & control
Gene Expression Regulation/drug effects
Glucose/immunology/*metabolism
Leupeptins/pharmacology
Male
Muscle, Smooth, Vascular/*cytology
Myocytes, Smooth Muscle/cytology/*drug effects/immunology/metabolism
NF-kappa B/*antagonists & inhibitors/immunology
Plasminogen Activator Inhibitor 1/*genetics
Proline/analogs & derivatives/pharmacology
Rats
Rats, Sprague-Dawley
Thiocarbamates/pharmacology