Urinary matrix metalloproteinase -8, -9, -14 and their regulators (TRY-1, TRY-2, TATI) in patients with diabetic nephropathy.
Matrix metalloproteinase-9 (MMP-9) has been shown to be involved in the development of diabetic nephropathy (DNP). We studied the levels, molecular forms, and degree of activation of urinary MMP-8, -9, -14, trypsin-1 and -2, as well as tumor-associated trypsin inhibitor (TATI) of DNP patients and healthy controls. Urinary samples were analyzed for MMPs by Western blotting and gelatin zymography and for trypsin-1, -2, and TATI by time-resolved immunofluorometric assays. Total MMP-8 immunoreactivity, the proportion of active MMP-9, and gelatinolytic activity in urine were significantly higher in DNP patients than in controls. In urine of DNP patients the proportion of active polymorphonuclear neutrophil (PMN)-type (but not fibroblast-type) MMP-8 was increased. MMP-8 and MMP-9 were found to form high molecular weight complexes in DNP urine. Total immunoreactivity of soluble urinary MMP-14 and the levels of trypsin (TRY)-1 and TRY-2, but not of TATI, were also significantly increased in DNP. Zymography, Western blotting, and immunofluorometric analysis of DNP urine showed a significant association especially between activation of MMP-9 as well as PMN-type MMP-8 and TRY-2. Our findings suggest that a trypsin-MMP cascade is involved in the pathogenesis of DNP, which may offer new possibilities for diagnosis and treatment of DNP with MMP inhibitors.
Lauhio
Anneli
A
Helsinki University Central Hospital, Department of Medicine, Division of Infectious Diseases, Helsinki, Finland. anneli.lauhio@hus.fi
Sorsa
Timo
T
Srinivas
Ravi
R
Stenman
Mathias
M
Tervahartiala
Taina
T
Stenman
Ulf-Håkan
UH
Grönhagen-Riska
Carola
C
Honkanen
Eero
E
D000328:Adult; D000368:Aged; D015153:Blotting, Western; D003928:Diabetic Nephropathies / Q000503:physiopathology*; D005260:Female; D015200:Fluoroimmunoassay; D006801:Humans; D008297:Male; D053511:Matrix Metalloproteinase 14 / Q000652:urine; D020784:Matrix Metalloproteinase 8 / Q000652:urine; D020780:Matrix Metalloproteinase 9 / Q000652:urine; D008875:Middle Aged; D013997:Time Factors; D014357:Trypsin / Q000378:metabolism*; D014359:Trypsin Inhibitor, Kazal Pancreatic / Q000378:metabolism*; D014362:Trypsinogen / Q000378:metabolism*
C064831:PRSS2 protein, human; D014359:Trypsin Inhibitor, Kazal Pancreatic; D014362:Trypsinogen; C489303:PRSS1 protein, human; D014357:Trypsin; D020784:Matrix Metalloproteinase 8; D020780:Matrix Metalloproteinase 9; D053511:Matrix Metalloproteinase 14