Background/Aims: Plasminogen activator inhibitor (PAI)-1 is increasingly recognized as a profibrotic factor but the mechanisms are not entirely clear. The present study examined the profibrotic mechanism of PAI-1 focusing on its effect on transforming growth factor (TGF)-β1 in experimental diabetes. Methods: PAI-1 knockout (KO) mesangial cells cultured under high glucose (HG) in addition to streptozotocin-induced diabetic PAI-1 KO mice were used. Results: PAI-1 deficiency did not affect plasma glucose significantly but reduced the fractional mesangial area, fibronectin and collagen I expression in the renal cortex after 20 weeks of diabetes as well as in HG-stimulated mesangial cells along with suppression of TGF-β1 mRNA expression. PAI-1 deficiency also reduced HG-induced βig-h3, a TGF-β1-induced gene product, mRNA expression. All these losses-of-function in PAI-1 KO mesangial cells were effectively gained by recombinant PAI-1. Recombinant PAI-1-induced fibronectin and collagen I expression was abrogated by TGF-β1 receptor inhibitor or anti-TGF-β antibody suggesting that the effect of PAI-1 was mediated by TGF-β1. In a similar context, recombinant PAI-1 stimulated TGF-β1 promoter activity to the same extent as TGF-β1 itself. Conclusion: Since TGF-β1 is well known to stimulate the PAI-1 promoter, we suggest that TGF-β1 and PAI-1 together constitute a positive feedback loop in the development of renal fibrosis in diabetes.

Keywords:
βig-h3, Diabetic nephropathy, Knockout mice, Matrix metalloproteinase, Mesangial cells, Plasmin, Plasminogen activator inhibitor-1, Renal fibrosis, Transforming growth factor-β1
1.
Mauer SM, Steffes MW, Ellis EN, Sutherland DE, Brown DM, Goetz FC: Structural-functional relationships in diabetic nephropathy. J Clin Invest 1984;74:1143–1155.
2.
Bilos R: Microvascular disease: what does the UKPDS tell us about diabetic nephropathy? Diabet Med 2008;25(suppl 2):25–29.
3.
Ingelfinger JR: Forestalling fibrosis. N Engl J Med 2003;349:2265–2266.
4.
Rerolle JP, Hertig A, Nguyen G, Sraer JD, Rondeau EP: Plasminogen activator inhibitor type 1 is a potential target in renal fibrogenesis. Kidney Int 2000;58:1841–1850.
5.
Eddy AA: Plasminogen activator inhibitor-1 and the kidney. Am J Physiol Renal Physiol 2002;283:F209–F220.
6.
Baricos WH, Cortez SL, el-Dahr SS, Schnaper HW: ECM degradation by cultured human mesangial cells is mediated by a PA/plasmin/MMP-2 cascade. Kidney Int 1995;47:1039–1047.
7.
Ramos-DeSimone N, Hahn-Dantona E, Sipley J, Nagase H, French DL, Quigley JP: Activation of matrix metalloproteinase-9 (MMP-9) via a converging plasmin/stromelysin-1 cascade enhances tumor cell invasion. J Biol Chem 1999;274:13066–13076.
8.
Monea S, Lehti K, Keski-Oja J, Mignatti P: Plasmin activates pro-matrix metalloproteinase-2 with a membrane-type 1 matrix metalloproteinase-dependent mechanism. J Cell Physiol 2002;192:160–170.
9.
Munger JS, Harpel JG, Gleizes PE, Mazzieri R, Nunes I, Rifkin DB: Latent transforming growth factor-beta: structural features and mechanisms of activation. Kidney Int 1997;51:1376–1382.
10.
Zhang G, Kernan KA, Collins SJ, Cai X, Lopez-Guisa JM, Degen JL, Shvil Y, Eddy AA: Plasmin(ogen) promotes renal interstitial fibrosis by promoting epithelial-to-mesenchymal transition: role of plasmin-activated signals. J Am Soc Nephrol 2007;18:846–859.
11.
Eddy AA, Fogo AB: Plasminogen activator inhibitor-1 in chronic kidney disease: evidence and mechanisms of action. J Am Soc Nephrol 2006;17:2999–3012.
12.
Lee EA, Seo JY, Jiang Z, Yu MR, Kwon MK, Ha H, Lee HB: Reactive oxygen species mediates HG-induced plasminogen activator inhibitor-1 up-regulation in mesangial cells and in diabetic kidney. Kindey Int 2005;67:1762–1771.
13.
Kitching AR, Holdsworth SR, Ploplis VA, Plow EF, Collen D, Carmeliet P, Tipping PG: Plasminogen and plasminogen activators protect against renal injury in crescentic glomerulonephritis. J Exp Med 1997;185:963–968.
14.
Kitching AR, Kong YZ, Huang XR, Davenport P, Edgtton KL, Carmeliet P, Holdsworth SR, Tipping PG: Plasminogen activator inhibitor-1 is a significant determinant of renal injury in experimental crescentic glomerulonephritis. J Am Soc Nephrol 2003;14:1487–1495.
15.
Oda T, Jung YO, Kim HS, Cai X, Lopez-Guisa JM, Ikeda Y, Eddy AA: PAI-1 deficiency attenuates the fibrogenic response to ureteral obstruction. Kidney Int 2001;60:587–596.
16.
Huang Y, Haraguchi M, Lawrence DA, Border WA, Yu L, Noble NA: A mutant, noninhibitory plasminogen activator inhibitor type 1 decreases matrix accumulation in experimental glomerulonephritis. J Clin Invest 2003;112:379–388.
17.
Krag S, Danielsen CC, Carmeliet P, Nyengaard J, Wogensen L: Plasminogen activator inhibitor-1 gene deficiency attenuates TGF-beta1-induced kidney disease. Kidney Int 2005;68:2651–2666.
18.
Matsuo S, López-Guisa JM, Cai X, Okamura DM, Alpers CE, Bumgarner RE, Peters MA, Zhang G, Eddy AA: Multifunctionality of PAI-1 in fibrogenesis: evidence from obstructive nephropathy in PAI-1-overexpression mice. Kidney Int 2005;67:2221–2238.
19.
Nicholas SB, Aguiniga E, Ren Y, Kim J, Wong J, Govindarajan N, Noda M, Wang W, Kawano Y, Collins A, Hsueh WA: Plasminogen activator inhibitor-1 deficiency retards diabetic nephropathy. Kidney Int 2005;67:1297–1307.
20.
Collins SJ, Alexander SL, López-Guisa JM, Cai X, Maruvada R, Chua SC, Zhang G, Okamura DM, Matsuo S, Eddy AA: Plasminogen activator inhibitor-1 deficiency has renal benefits but some adverse systemic consequences in diabetic mice. Nephron Exp Nephrol 2006;104:e23–e34.
21.
Lassila M, Fukami K, Jandeleit-Dahm K, Semple T, Carmeliet P, Cooper ME, Kitching AR: Plasminogen activator inhibitor-1 production is pathogenetic in experimental murine diabetic renal disease. Diabetologia 2007;50:1315–1326.
22.
Huang Y, Border WA, Yu L, Zhang J, Lawrence DA, Noble NA: A PAI-1 mutant, PAI-1R, slows progression of diabetic nephropathy. J Am Soc Nephrol 2008;19:329–338.
23.
Ha H, Oh E, Lee HB: The role of plasminogen activator inhibitor1 in renal and cardiovascular diseases. Nat Rev Nephrol 2009;5:203–211.
24.
Gilbert RE, Wilkinson-Berka JL, Johnson DW, Cox A, Soulis T, Wu LL, Kelly DJ, Jerums G, Pollock CA, Cooper ME: Renal expression of TGF-β inducible gene-h3 (beta ig-h3) in normal and diabetic rats. Kidney Int 1998;54:1052–1062.
25.
Jiang Z, Seo JY, Ha H, Lee EA, Kim YS, Han DC, Uh ST, Park CS, Lee HB: Reactive oxygen species mediate TGF-β1-induced plasminogen activator inhibtor-1 upregulation in mesangial cells. Biochem Biophys Res Commun 2003;309:961–966.
26.
Moon JA, Kim HT, Cho IS, Sheen YY, Kim DK: IN-1130, a novel transforming growth factor-β type1 receptor kinase (ALK5) inhibitor, suppresses renal fibrosis in obstructive nephropathy. Kidney Int 2006;70:1234–1243.
27.
Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248–254.
28.
Noh H, Kim JS, Han K-H, Lee GT, Song JS, Chung SH, Jeon JS, Ha H, Lee HB: Oxidative stress during peritoneal dialysis: implications in functional and structural changes in the membrane. Kidney Int 2006;69:2022–2028.
29.
Kim SJ, Glick A, Sporn MB, Roberts AB: Characterization of the promoter region of the human transforming growth factor-beta 1 gene. J Biol Chem 1989;264:402–408.
30.
Kim SJ, Jeang KT, Glick AB, Sporn MB, Roberts AB: Promoter sequences of the human transforming growth factor-beta 1 gene responsive to transforming growth factor-beta 1 autoinduction. J Biol Chem 1989;264:7041–7045.
31.
Lee HB, Yu MR, Song JS, Ha H: Reactive oxygen species amplify protein kinase C signaling in HG-induced fibronectin expression by peritoneal mesothelial cells. Kidney Int 2004;65:1170–1179.
32.
Okada Y, Morodomi T, Enghild JJ, Suzuki K, Yasui A, Nakanishi I, Salvesen G, Nagase H: Matrix metalloproteinase 2 from human rheumatoid fibroblasts. Purification and activation of the precursor and enzymatic properties. Eur J Biochem 1990;194:721–730.
33.
Liotta LA, Goldfarb RH, Brundage R, Siegal GP, Terranova V, Garbisa S: Effect of plasminogen activator (urokinase), plasmin, and thrombin on glycoprotein and collagenous components of basement membrane. Cancer Res 1981;41:4629–4636.
34.
Lee HB, Seo J, Jiang Z, Ha H: Suppression of plasminogen activator inhibitor-1 inhibits HG- and TGF-β1-induced fibronectin secretion and increases MMP2 activity by mesangial cells (abstract). J Am Soc Nephrol 2002;13:169A.
35.
Baricos WH, Cortez SL, Deboisblanc M, Xin S: Transforming growth factor-beta is a potent inhibitor of extracellular matrix degradation by cultured human mesangial cells. J Am Soc Nephrol 1999;10:790–795.
36.
Mclennan SV, Fisher E, Martell SY, Death AK, Williams PF, Lyons JG, Yue DK: Effects of glucose on matrix metalloproteinase and plasmin activities in mesangial cells: possible role in diabetic nephropathy. Kidney Int Suppl 2000;77:S81–S87.
37.
Baricos WH, Reed JC, Cortez SL: Extracellular matrix degradation by cultured mesangial cells: mediators and modulators. Exp Biol Med (Maywood) 2003;228:1018–1022.
38.
Wong AP, Cortez SL, Baricos WH: Role of plasmin and gelatinase in extracellular matrix degradation by cultured rat mesangial cells. Am J Physiol 1992;263:F1112–F1118.
39.
Hertig A, Berrou J, Allory Y, Breton L, Commo F, Costa De Beauregard MA, Carmeliet P, Rondeau E: Type 1 plasminogen activator inhibitor deficiency aggravates the course of experimental glomerulonephritis through overactivation of transforming growth factor β. FASEB J 2003;17:1904–1906.
40.
Pedroja BS, Kang LE, Imas AO, Carmeliet P, Bernstein AM: Plasminogen activator inhibitor-1 regulates integrin αvβ3 expression and autocrine TGFβ signaling. J Biol Chem 2009;284:20708–20717.
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2009
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