Plasminogen Activator Inhibitor-1 Antisense Oligodeoxynucleotides Abrogate Mesangial Fibronectin Accumulation.
10.4196/kjpp.2010.14.6.385
- Author:
Jehyun PARK
1
;
Ji Yeon SEO
;
Hunjoo HA
Author Information
1. Department of Bioinspired Science, Division of Life and Pharmaceutical Sciences, College of Pharmacy, Ewha Womans University, Seoul 120-752, Korea. hha@ewha.ac.kr
- Publication Type:Original Article
- Keywords:
Plasminogen activator inhibitor-1;
Antisense oligodeoxynucleotide;
Plasmin;
Matrix metalloproteinase;
Mesangial cells
- MeSH:
Animals;
Blotting, Western;
Diabetic Nephropathies;
Extracellular Matrix;
Fibrinolysin;
Fibronectins;
Fibrosis;
Glucose;
Liposomes;
Mesangial Cells;
Oligodeoxyribonucleotides;
Plasminogen;
Plasminogen Activator Inhibitor 1;
Plasminogen Activators;
Rats;
Renal Insufficiency, Chronic;
Transfection;
Transforming Growth Factor beta1;
Transforming Growth Factors;
Up-Regulation
- From:The Korean Journal of Physiology and Pharmacology
2010;14(6):385-390
- CountryRepublic of Korea
- Language:English
-
Abstract:
Excessive extracellular matrix (ECM) accumulation is the main feature of chronic renal disease including diabetic nephropathy. Plasminogen activator inhibitor (PAI)-1 is known to play an important role in renal ECM accumulation in part through suppression of plasmin generation and matrix metalloproteinase (MMP) activation. The present study examined the effect of PAI-1 antisense oligodeoxynucleotide (ODN) on fibronectin upregulation and plasmin/MMP suppression in primary mesangial cells cultured under high glucose (HG) or transforming growth factor (TGF)-beta1, major mediators of diabetic renal ECM accumulation. Growth arrested and synchronized rat primary mesangial cells were transfected with 1 microM phosphorothioate-modified antisense or control mis-match ODN for 24 hours with cationic liposome and then stimulated with 30 mM D-glucose or 2 ng/ml TGF-beta1. PAI-1 or fibronectin protein was measured by Western blot analysis. Plasmin activity was determined using a synthetic fluorometric plasmin substrate and MMP-2 activity analyzed using zymography. HG and TGF-beta1 significantly increased PAI-1 and fibronectin protein expression as well as decreased plasmin and MMP-2 activity. Transient transfection of mesangial cells with PAI-1 antisense ODN, but not mis-match ODN, effectively reversed basal as well as HG- and TGF-beta1-induced suppression of plasmin and MMP-2 activity. Both basal and upregulated fibronectin secretion were also inhibited by PAI-1 antisense ODN. These data confirm that PAI-1 plays an important role in ECM accumulation in diabetic mesangium through suppression of protease activity and suggest that PAI-1 antisense ODN would be an effective therapeutic strategy for prevention of renal fibrosis including diabetic nephropathy.
