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Effect of heparin on high glucose induced proliferation and expression of matrix metalloproteinases in normal human mesangial cells

  • Zhou Qiao-ling 
  • Yasumoto Yuichiro
  • Tsukamoto Masatoshi
  • Nozaki Tsuyoshi
  • Sogabe Atsushi
  • Harada Kouji
  • Zhang Yi-xiang 
  • Lin Xiao-yan 
  • Zhang Yang-de 
  • Arima Terukatsu
Medicine

Abstract

Background The pathogenesis of diabetic nephropathy (DN) is a complex pathophysiological process. Its precise mechanism is not fully known. In recent years it has been recognized that synthesis of various extracelluar matrix (ECM) components may increase, and that degradation of ECM may decrease in DN. It was reported heparin could inhibit mesangial cells proliferation in vitro. The main aim of this study is to explore whether heparin inhibits proliferation of mesangial cells grown in high glucose concentration and to measure the effect of heparin on matrix metalloproteinases (MMPs) expression in mesangial cells. Methods The medium contained either low glucose (5 mmol/L) or high glucose (25 mmol/L). The concentrations of heparin in the culture medium were 0, 25, 50, 100, 200 or 400 µg/mL. A metabolic (WST-1) assay was used to measure mesangial cell proliferation and Western blot analysis was used to measure MMPs expression of mesangial cells. Results Normal human mesangial cell (NHMC) proliferation was higher in high glucose (HG) medium than in low glucose (LG) medium. They showed a 1.93 fold expansion after 72 h in high glucose in contrast to a 1.63 fold expansion in low glucose. In the presence of heparin, mesangial cells proliferation was inhibited, which was more obvious at high glucose concentrations than at low glucose concentrations. In high glucose, with heparin concentration of 50, 100, 200 and 400 µg/mL, the mesangial cells showed a 0.61 fold, 0.52 fold, 0.52 fold and 0.41 fold reductions in cell number compared to cells grown without heparin. In low glucose, only concentrations of 200 µg/mL and 400 µg/mL showed reduction in cell number, namely 0.54 fold and 0.45 fold, when compared to cells grown without heparin. In Western blot analysis, MMP1, MMP2, MMP3 and MMP9 was expressed by mesangial cells expressed in both high and low glucose concentrations, which was more prominent in high glucose medium. Incubation of heparin further increased expression of MMP1, MMP2, MMP3 and MMP9. Conclusions This study suggests that glucose can accelerate mesangial cell proliferation while heparin can reduce proliferation, being more obvious at high glucose concentrations. Higher glucose concentrations led to increased MMP expression, which may take part in the regulation of mesangial matrix synthesis and degradation. Addition of heparin resulted in a corresponding increase in MMP expression, most notably at high glucose concentrations, indicating a potentially renoprotective role in DN.

Key words

matrix metalloproteinases normal human mesangial cell glucose heparin 

CLC number

R587.1 

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References

  1. [1]
    Bails B K. Diabetes mellitus and its chronic complications[J]. Aorn J, 2002, 76(2): 266–276.Google Scholar
  2. [2]
    Lee T S, Saltsman K A, Ohashi H, et al. Activation of protein in kinase C by elevation of glucose concentration: Proposal for a mechanism in the development of diabetic vascular complications[J]. Proc Natt Acad Sci USA, 1989, 86: 5141–5145.CrossRefGoogle Scholar
  3. [3]
    Thomas Stearns Eliot. Incidence and prevalence of ESRD [J]. J Am Kid Dise, 2001, 38(4): S37–52.Google Scholar
  4. [4]
    Brenner B M, Anderson S. Glomerular function in diabetes mellitus[J]. Adv Nephrol, 1990, 19: 135–144.Google Scholar
  5. [5]
    Greene D A, Lattimer S A, Sima A. Sorbitol, phospho in ositides, and sodium-potassium-ATPase in the pathogenesis of diabetic complications[J]. N Engl J Med, 1987, 316: 599–606.CrossRefGoogle Scholar
  6. [6]
    Horie K, Miyata T, Maeda K, et al. Immunohistochemical colocalization of glycoxidation products and Lipid peroxidation products in diabetic renal glomerular lesions[J]. J Clin Invest, 1997, 100: 2995–3004.CrossRefGoogle Scholar
  7. [7]
    Abboud H E. Grouth factors and diabetic nephropathy: An overview [J]. Kidney Int, 1997, 52(S60): 3–6.Google Scholar
  8. [8]
    Ayo S H, Radnik R A, Glass W F II. Increased extracellar matrix synthesis and mRNA in mesangial cells grown in high glucose medium [J]. Am J Physiol, 1991, 260: F181–191.CrossRefGoogle Scholar
  9. [9]
    Sharma L, Ziyadeh F N. Hyperglycemia and diabetic kidney disease[J]. Diabetes, 1995, 44: 1139–1146.CrossRefGoogle Scholar
  10. [10]
    Fisher E, Mclennan S, Tada H, et al. Interaction of ascorbic acid and glucose on the production of collagen and proteoglycan by fibroblasts [J]. Diabetes, 1991, 40: 371–376.CrossRefGoogle Scholar
  11. [11]
    Pugliese G, Pricci F, Pugliese F, et al. Mechanisms of glucose enhanced extracellular matrix accumulation in rat glomerular mesangial cells[J]. Diabetes, 1994, 43: 478–490.CrossRefGoogle Scholar
  12. [12]
    Anderson S S, Wu K J, Nagase H, et al. Effect of matrix glycation on expression of typelv Collagen, MMP2, MMP9 and TIMP-1, by human mesangial cells[J]. Cell Adhesion and Communication, 1996, 4: 89–101.CrossRefGoogle Scholar
  13. [13]
    Steffes M W, Osterby R, Chavers B, et al. Mesangial expansion as a central mechanism for loss of kidney function in diabetic patients[J]. Diabetes, 1989, 38: 1077–1081.CrossRefGoogle Scholar
  14. [14]
    Mclennan S, Fisher E, Yue D K, et al. High glucose concentration causes a decrease in mesangium degradation[J]. Diabetes, 1994, 43: 1041–1045.CrossRefGoogle Scholar
  15. [15]
    Mclennan S, Martell S Y, Yue D K. High glucose concentration inhibits the expression of membrane type metalloproteinase by mesangial cells: Possible role in mesangium accumulation [J]. Diabetologia, 2000, 43: 642–648.CrossRefGoogle Scholar
  16. [16]
    Steffes M W, Bilous R W, Sutherland D E R, et al. Cell and matrix components of the glomerular mesangium in type I diabetes[J]. Diabetes, 1992, 41: 679–684.CrossRefGoogle Scholar
  17. [17]
    Suzuki D. Metalloproteinases in the pathogenesis of diabetic nephropathy[J]. Nephron, 1998, 80: 125–133.CrossRefGoogle Scholar
  18. [18]
    Mclennan S, Fisher E, Martell S Y, et al. Effects of glucose on matrix metalloproteinase and plasmin activities in mesangial cells: Possible role in diabetic nephropathy[J]. Kidney Int, 2000, 58(S177): 81–87.CrossRefGoogle Scholar
  19. [19]
    Striker L J. Peten E P, Elliot S J, et al. Mesangial cell turnerover effect of heparin and peptide growth factor[J]. Lab Invest. 1991, 64: 446–456.Google Scholar
  20. [20]
    Castellot J J, Hoover R L, Harper P A, et al. Heparin and glomerular epithelial cell-secreted heparin-like species inhibit mesangial cell proliferation[J]. Am J Pathol, 1985, 120: 427–435.Google Scholar
  21. [21]
    Castellot J J, Hoover R L, Karnovsky M J. Glomerular endothelial cells secrete a heparin-like inhibitor and a peptide stimulator of mesangial cell proliferation [J]. Am J Pathol, 1986, 125: 493–500.Google Scholar
  22. [22]
    Groggel G C, Marinidez G N, Hovingh P, et al. Inhibition of rat mesangial cell growth by heparin sulfate[J]. Am J Physiol, 1990, 258: F259–265.Google Scholar
  23. [23]
    Kreisberg J I, Kreisberg S H, High glucose activates PKC and stimulates fibronectin gene expression by enhancing a cAMP response element[J]. Kidney Int, 1995, 48(S 51): S3-S11.Google Scholar
  24. [24]
    Yokoyama H, Deckert T. Central role of TGF-β in the pathogenesis of diabetic nephropathy and macro vascular complications[J]. Diabet Med, 1996, 13: 313–320CrossRefGoogle Scholar
  25. [25]
    Studer R K, Craven P A, Derbrtis F R. Role for protein kinase C in the mediation of increased fibronectin accumulation by mesangial cells grown in high glucose medium[J]. Am J Pathol, 1989, 134: 843–855.Google Scholar
  26. [26]
    Handa M, Araki S, Togawa M, et al. Mitogen-activated-protein kinase cascade is activated in glomeruli of diabetic rats and glomerular mesangial cells cultured under high glucose conditions [J]. Diabetes, 1997, 46: 847–853.CrossRefGoogle Scholar
  27. [27]
    Crowley S T, Brownlee M, Edelstein D, et al. Effects of no nenzymatic glycosylation of matrix on proliferation of mesangial cells[J]. Diabetes, 1991, 40: 540–547.CrossRefGoogle Scholar
  28. [28]
    Ziyadeh F N, Sharma K, Ericksen M, et al. Stimulation of collagen gene expression and protein synthesis in marine mesangial cells by high glucose is mediated by autocrine activation of transforming growth factor β[J]. J Clin Invest, 1994, 93: 536–542.CrossRefGoogle Scholar
  29. [29]
    Barics W H, Shah S V. Proteolytic enzymes mediators of glomerular injury[J]. Kidney Int, 1991, 40: 161–173.CrossRefGoogle Scholar
  30. [30]
    Purkerson J M, Joist J H, Greenberg P J M, et al. Inhibition by anticoagulant drugs of the progressive hypertension and uremia associated with renal infarction in rats[J]. Thromb Res, 1982, 26: 227–240.CrossRefGoogle Scholar
  31. [31]
    Olson J L. Role of heparin as a protective agent following reduction of renal mass[J]. Kidney Int, 1984, 25: 376–382.CrossRefGoogle Scholar
  32. [32]
    Benador N M, Dirardin E P. Influence of heparin and type-IV collagen on IL-6 synthesis by rat glomerular mesangial cells[J]. Nephron, 1997, 77: 219–224.CrossRefGoogle Scholar
  33. [33]
    Ceol M, Baggio B, Gambaro G, et al: Effect of heparin treatment on glomerular collagen IV alpha1 and TGF-b1 expression and deposition in diabetic rats[J]. Diabetologia, 1995, 38(S1): 62.Google Scholar

Copyright information

© Central South University 2005

Authors and Affiliations

  • Zhou Qiao-ling 
    • 1
    • 2
  • Yasumoto Yuichiro
    • 1
  • Tsukamoto Masatoshi
    • 1
  • Nozaki Tsuyoshi
    • 1
  • Sogabe Atsushi
    • 1
  • Harada Kouji
    • 1
  • Zhang Yi-xiang 
    • 1
  • Lin Xiao-yan 
    • 1
  • Zhang Yang-de 
    • 2
  • Arima Terukatsu
    • 1
  1. 1.Second Department of Internal MedicineKagoshima UniversityKagoshimaJapan
  2. 2.Department of Nephrology, Xiangya HospitalCentral South UniversityChangshaChina

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