Mechanism of matrix accumulation and glomerulosclerosis in spontaneously hypertensive rats.

Abstract

Renal interstitial fibrosis and thickening of the glomerular basement membrane are associated with hypertension. However, the mechanism of matrix accumulation is unclear. Spontaneously hypertensive rats (SHR) develop hypertension at between 2 and 6 weeks of age. To test the hypothesis that increased matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) contribute to the pathomechanisms of hypertensive nephropathy, the cortex and medulla of male SHR at 2 and 6 weeks were analyzed for MMP-2, MMP-7, and MMP-9 by gelatin and elastin gel zymography. The levels of TIMP-4 were measured by western blot analysis. The bands in blots were scanned and normalized with actin. To localize MMP-2 and TIMP-4 in situ, immuno-labeling was performed. To determine proteinuresis, urinary protein was measured by Bio-Rad dye binding assay. The mean arterial pressure (mmHg) was measured in Inactin-anesthetized rats by a PE-50 catheter in the femoral artery. Age-sex matched normotensive Wistar rats (NWR) were used as controls and grouped: (1). SHR, 2 weeks; (2). SHR, 6 weeks; (3). NWR, 2 weeks; and (4). NWR, 6 weeks (n = 6 in each group). Levels of cortex MMP-2 and MMP-9 were increased in 6 week SHR as compared with NWR. In the medulla, MMP-9 and MMP-7 were increased, but there was no change in MMP-2. The levels of cortex TIMP-4 tended to increase but insignificantly. In contrast, there were significant increases in the levels of TIMP-4 in the medulla of 6 week SHR as compared with 2 week SHR or NWR. In addition, there were substantial elastinolytic activity in the cortex of 6 week SHR. The in situ labeling suggested no TIMP-4 in the glomeruli. There was substantial TIMP-4 in the epithelial layer of tubules. The levels of fibrotic collagen were significantly higher in both the glomeruli and tubular interstitium. Urinary protein excretion was increased significantly in 6 week SHR when compared with other groups. The mean arterial pressure was 1.6-fold higher in 6 week SHR than in controls. These results suggest that increased MMP-2 and MMP-9 activity contributes to glomerular injury and hypertensive remodeling. The increased levels of TIMP-4 in the medulla may inhibit the collagenolytic activity of MMP but is unable to inhibit the elastinolytic activity. An important role of MMP-2, MMP-9, and TIMP-4 in hypertensive remodeling of the cortex and medulla in the SHR is demonstrated.