Expression profile of extracellular matrix and its regulatory proteins during the process of interstitial fibrosis after anti-glomerular basement membrane antibody-induced glomerular sclerosis in Sprague-Dawley rats

1999 ◽  
Vol 49 (8) ◽  
pp. 716-725 ◽  
Author(s):  
Laxman P. Adhikary ◽  
Tadashi Yamamoto ◽  
Masato Isome ◽  
Yoshimasa Nakano ◽  
Katsutoshi Kawasaki ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Basement Membrane ◽  
Expression Profile ◽  
Glomerular Basement Membrane ◽  
Interstitial Fibrosis ◽  
Regulatory Proteins ◽  
Glomerular Sclerosis ◽  
Sprague Dawley ◽  
Sprague Dawley Rats

scholarly journals Degradation of glomerular basement membrane by purified mammalian metalloproteinases

1988 ◽  
Vol 254 (2) ◽  
pp. 609-612 ◽  
Author(s):  
W H Baricos ◽  
G Murphy ◽  
Y W Zhou ◽  
H H Nguyen ◽  
S V Shah
Keyword(s):  
Extracellular Matrix ◽  
Basement Membrane ◽  
Glomerular Basement Membrane ◽  
Degradation Products ◽  
Gel Chromatography ◽  
Collagen Types ◽  
Percentage Release ◽  
Dose Dependent

Neutral metalloproteinases degrade components of the extracellular matrix, including collagen types I-V, fibronectin, laminin and proteoglycan. However, their ability to degrade intact glomerular basement membrane (GBM) has not previously been investigated. Incubation of [3H]GBM (50,000 c.p.m.; pH 7.5; 24 h at 37 degrees C) with purified gelatinase or stromelysin (2 units) resulted in significant GBM degradation: gelatinase, 46 +/- 2.2; stromelysin, 59 +/- 5.8 (means +/- S.E.M.; percentage release of non-sedimentable radioactivity; n = 4). In contrast, 2 units of collagenase released only 5.6 +/- 0.52% (n = 3) of the [3H]GBM radioactivity compared with 2.0 +/- 0.15% (n = 7) released from [3H]GBM incubated alone. Sephadex G-200 gel chromatography of supernatants obtained from incubations of [3H]GBM with either gelatinase or stromelysin confirmed the ability of these enzymes to degrade GBM and revealed both high-(800,000) and relatively low-(less than 20,000) Mr degradation products for both enzymes. GBM degradation by gelatinase and stromelysin was dose-dependent (range 0.02-2.0 units), near maximal between pH 6.0 and 8.6, and was completely inhibited (greater than 95%) by 2 mM-o-phenanthroline. Collagenase (2 units) did not enhance the degradation of GBM by either gelatinase (0.02 or 0.2 unit) or stromelysin (0.02 or 0.2 unit). Our results indicate that metalloproteinase-mediated GBM degradation by neutrophils and glomeruli may be attributable to gelatinase (neutrophils) and/or stromelysin (glomeruli) and suggest an important role for these proteinases in glomerular pathophysiology.

scholarly journals Asymmetric dimethylarginine in angiotensin II-induced hypertension

2010 ◽  
Vol 298 (3) ◽  
pp. R740-R746 ◽  
Author(s):  
Jennifer M. Sasser ◽  
Natasha C. Moningka ◽  
Mark W. Cunningham ◽  
Byron Croker ◽  
Chris Baylis
Keyword(s):  
Nitric Oxide ◽  
Renal Injury ◽  
Asymmetric Dimethylarginine ◽  
Renal Cortex ◽  
Interstitial Fibrosis ◽  
Glomerular Sclerosis ◽  
Ang Ii ◽  
Sprague Dawley ◽  
Plasma Adma ◽  
Induced Hypertension

Recent studies have shown that asymmetric dimethylarginine (ADMA), a nitric oxide synthase inhibitor, is increased in hypertension and chronic kidney disease. However, little is known about the effects of hypertension per se on ADMA metabolism. The purpose of this study was to test the hypothesis that ANG II-induced hypertension, in the absence of renal injury, is associated with increased oxidative stress and plasma and renal cortex ADMA levels in rats. Male Sprague-Dawley rats were treated with ANG II at 200 ng·kg−1·min−1 sc (by minipump) for 1 or 3 wk or at 400 ng·kg−1·min−1 for 6 wk. Mean arterial pressure was increased after 3 and 6 wk of ANG II; however, renal injury (proteinuria, glomerular sclerosis, and interstitial fibrosis) was only evident after 6 wk of treatment. Plasma thiobarbituric acid reactive substances concentration and renal cortex p22phox protein abundance were increased early (1 and 3 wk), but urinary excretion of isoprostane and H2O2 was only increased after 6 wk of ANG II. An increased in plasma ADMA after 6 wk of ANG II was associated with increased lung protein arginine methyltransferase-1 abundance and decreased renal cortex dimethylarginine dimethylaminohydrolase activity. No changes in renal cortex ADMA were observed. ANG II hypertension in the absence of renal injury is not associated with increased ADMA; however, when the severity and duration of the treatment were increased, plasma ADMA increased. These data suggest that elevated blood pressure alone, for up to 3 wk, in the absence of renal injury does not play an important role in the regulation of ADMA. However, the presence of renal injury and sustained hypertension for 6 wk increases ADMA levels and contributes to nitric oxide deficiency and cardiovascular disease.

Pathomorphological Sequence of Nephron Loss in Diabetic Nephropathy

2021 ◽  
Author(s):  
Jana Löwen ◽  
Elisabeth Gröne ◽  
Marie-Luise Groß-Weißmann ◽  
Felix Bestvater ◽  
Hermann-Josef Gröne ◽  
...  
Keyword(s):  
Diabetic Nephropathy ◽  
Basement Membrane ◽  
Interstitial Fibrosis ◽  
Glomerular Sclerosis ◽  
Structural Aberrations ◽  
Vascular Proliferation ◽  
Advanced Stages ◽  
Nephron Loss ◽  
Bowman's Capsule ◽  
Glomerular Capillaries

Abstract Following our reports on mesangial sclerosis and vascular proliferation in diabetic nephropathy (DN)(25,34) we now describe the advanced stages of DN terminating in glomerular obsolescence and tubulo-interstitial fibrosis based on a total of 918 biopsies. The structural aberrations emerge from two defects: First, an increased synthesis of glomerular basement membrane (GBM) components by podocytes and endothelial cells leading to an accumulation of GBM material in the mesangium. Second, a defect of glomerular vessels consisting of an increased leakiness and an increased propensity to proliferate. Both defects may lead to glomerular degeneration. The progressing compaction of the accumulated worn-out GBM-material together with the retraction of podocytes out of the tuft and the collapse and hyalinosis of capillaries results in a shrunken tuft that fuses with Bowman's capsule to glomerular sclerosis. The most frequent pathway to glomerular decay starts with local tuft expansions that result in contacts of structurally healthy podocytes to the parietal epithelium initiating the formation of tuft adhesions, which include the penetration of glomerular capillaries into BC. Exudation of plasma from such capillaries into the space between the parietal epithelium and its basement membrane causes the formation of insudative fluid accumulations within BC spreading around the glomerular circumference and, via the glomerulo-tubular junction, onto the tubule. Degeneration of the corresponding tubule develops secondarily to the glomerular damage, either due to cessation of filtration in cases of global sclerosis or due to encroachment of the insudative spaces. The degenerating tubules induce the proliferation of myo-fibroblasts resulting in interstitial fibrosis.

scholarly journals Localization of α-Dystroglycan on the Podocyte: from Top to Toe

2005 ◽  
Vol 53 (11) ◽  
pp. 1345-1353 ◽  
Author(s):  
Nils P.J. Vogtländer ◽  
Henry Dijkman ◽  
Marinka A.H. Bakker ◽  
Kevin P. Campbell ◽  
Johan van der Vlag ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Monoclonal Antibodies ◽  
Confocal Microscopy ◽  
Cell Membrane ◽  
Basement Membrane ◽  
Immunoelectron Microscopy ◽  
Glomerular Basement Membrane ◽  
Human Kidney ◽  
Charged Membrane ◽  
Apical Localization

α-Dystroglycan (DG) is a negatively charged membrane-associated glycoprotein that links the cytoskeleton to the extracellular matrix. Previously, we described that α-DG covers the whole podocyte cell membrane in the rat. However, our finding was challenged by the description of a strictly basolateral localization in human kidney biopsies, using a different antibody against α-DG. Therefore, we studied the exact localization of glomerular α-DG by using these two antibodies in both species. The studies were performed by using monoclonal antibodies (MoAbs) IIH6 and VIA4.1 in immunofluorescence, confocal microscopy, and immunoelectron microscopy on both rat and human kidney sections, as well as on cultured mouse podocytes. The apical localization of α-DG on podocytes was more dominant than the basolateral localization. The basolateral staining with MoAb VIA4.1 was more pronounced than that of MoAb IIH6. With both MoAbs, the staining in rat kidneys was more prominent, in comparison to human kidneys. We conclude that α-DG is expressed at both the basolateral and apical sides of the podocyte. This localization suggests that α-DG plays a dual role in the maintenance of the unique architecture of podocytes by its binding to the glomerular basement membrane, and in the maintenance of the integrity of the filtration slit, respectively.

17β-Estradiol supplementation reduces tubulointerstitial fibrosis by increasing MMP activity in the diabetic kidney

2007 ◽  
Vol 292 (2) ◽  
pp. R769-R777 ◽  
Author(s):  
Richard W. Mankhey ◽  
Corinne C. Wells ◽  
Faizah Bhatti ◽  
Christine Maric
Keyword(s):  
Extracellular Matrix ◽  
Diabetic Nephropathy ◽  
Tubulointerstitial Fibrosis ◽  
Relative Optical Density ◽  
Sprague Dawley ◽  
Type Iv ◽  
Sprague Dawley Rats ◽  
Ecm Degradation ◽  
17Β Estradiol ◽  
Renal Expression

We previously reported that supplementation with 17β-estradiol (E2) attenuates albuminuria, glomerulosclerosis, and tubulointerstitial fibrosis in diabetic nephropathy. The present study examined the mechanisms by which E2 regulates extracellular matrix (ECM) metabolism, a process that contributes to the development of glomerulosclerosis and tubulointerstitial fibrosis. The study was performed in female nondiabetic (ND), streptozotocin-induced diabetic (D), and diabetic with E2 supplementation (D+E2) Sprague-Dawley rats for 12 wk. Diabetes was associated with an increase in the renal expression of collagen α type IV [ND, 0.22 ± 0.02; D, 0.99 ± 0.09 relative optical density (ROD); P < 0.05] and fibronectin protein (ND, 0.36 ± 0.08; D, 1.47 ± 0.08 ROD; P < 0.05), as measured by Western blot analysis. E2 supplementation partially attenuated this increase in collagen α type IV (D+E2, 0.47 ± 0.10 ROD) and fibronectin (D+E2, 0.71 ± 0.16 ROD) protein expression associated with D. Diabetes was also associated with a decrease in the expression of matrix metalloproteinase (MMP) isoform MMP-2 (ND, 0.79 ± 0.01; D, 0.62 ± 0.06 ROD; P < 0.05) and MMP-9 protein (ND, 0.49 ± 0.02; D, 0.33 ± 0.03 ROD; P < 0.05). E2 supplementation restored MMP-2 and MMP-9 protein to levels similar or even greater than in the ND kidneys (MMP-2, 0.75 ± 0.06; MMP-9, 0.73 ± 0.01 ROD). The activities of MMP-2 (ND, 7.88 ± 0.44; D, 5.60 ± 0.54 ROD; P < 0.05) and MMP-9 (ND, 29.9 ± 1.8; D, 12.9 ± 2.3 ROD; P < 0.05), as measured by zymography, were also decreased with D. E2 supplementation restored MMP-2 and MMP-9 activity to levels similar to that in ND kidneys (MMP-2, 7.66 ± 0.35; MMP-9, 21.4 ± 2.9 ROD). We conclude that E2 supplementation is renoprotective by attenuating glomerulosclerosis and tubulointerstitial fibrosis by reducing ECM synthesis and increasing ECM degradation.

Phosphate overload induces podocyte injury via type III Na-dependent phosphate transporter

2011 ◽  
Vol 300 (4) ◽  
pp. F848-F856 ◽  
Author(s):  
Sahoko Sekiguchi ◽  
Atsushi Suzuki ◽  
Shogo Asano ◽  
Keiko Nishiwaki-Yasuda ◽  
Megumi Shibata ◽  
...  
Keyword(s):  
Basement Membrane ◽  
Glomerular Basement Membrane ◽  
Connexin 43 ◽  
Chronic Renal Disease ◽  
Rat Kidney ◽  
Cellular Damage ◽  
Glomerular Sclerosis ◽  
Wild Type ◽  
Podocyte Injury ◽  
Type Iii

Uptake of Pi at the cellular membrane is essential for the maintenance of cell viability. However, phosphate overload is also stressful for cells and can result in cellular damage. In the present study, we investigated the effects of the transgenic overexpression of type III Pi transporter Pit-1 to explore the role of extracellular Pi in glomerular sclerosis during chronic renal disease. Pit-1 transgenic (TG) rats showed progressive proteinuria associated with hypoalbuminemia and dyslipidemia. Ultrastructural analysis of TG rat kidney by transmission electron microscopy showed a diffuse effacement of the foot processes of podocytes and a thickening of the glomerular basement membrane, which were progressively exhibited since 8 wk after birth. TG rats died at 32 wk of age due to cachexia. At this time, more thickening of the glomerular basement membrane and segmental sclerosis were observed in glomeruli of the TG rats. Immunohistochemical examination using anti-connexin 43 and anti-desmin antibodies suggested the progressive injury of podocytes in TG rats. TG rats showed higher Pi uptake in podocytes than wild-type rats, especially under low Pi concentration. When 8-wk-old wild-type and TG rats were fed a 0.6% normal phosphate (NP) or 1.2% phosphate (HP) diet for 12 wk, HP diet-treated TG rats showed more progressive proteinuria and higher serum creatinine levels than NP diet-treated TG rats. In conclusion, our findings suggest that overexpression of Pit-1 in rats induces phosphate-dependent podocyte injury and damage to the glomerular barrier, which result in the progression of glomerular sclerosis in the kidney.

scholarly journals Analysis of Chemokines and Receptors Expression Profile in the Myelin MutantTaiepRat

2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Guadalupe Soto-Rodriguez ◽  
Juan-Antonio Gonzalez-Barrios ◽  
Daniel Martinez-Fong ◽  
Victor-Manuel Blanco-Alvarez ◽  
Jose R. Eguibar ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Expression Profile ◽  
Proinflammatory Cytokines ◽  
Mrna Levels ◽  
Pcr Array ◽  
Sprague Dawley ◽  
Protein Levels ◽  
Chronic Neuroinflammation ◽  
Sprague Dawley Rats ◽  
Old Rats

Taieprat has a failure in myelination and remyelination processes leading to a state of hypomyelination throughout its life. Chemokines, which are known to play a role in inflammation, are also involved in the remyelination process. We aimed to demonstrate that remyelination-stimulating factors are altered in the brainstem of 1- and 6-month-oldtaieprats. We used a Rat RT2Profiler PCR Array to assess mRNA expression of 84 genes coding for cytokines, chemokines, and their receptors. We also evaluated protein levels of CCL2, CCR1, CCR2, CCL5, CCR5, CCR8, CXCL1, CXCR2, CXCR4, FGF2, and VEGFA by ELISA. Sprague-Dawley rats were used as a control. PCR Array procedure showed that proinflammatory cytokines were not upregulated in thetaieprat. In contrast, some mRNA levels of beta and alpha chemokines were upregulated in 1-month-old rats, but CXCR4 was downregulated at their 6 months of age. ELISA results showed that CXCL1, CCL2, CCR2, CCR5, CCR8, and CXCR4 protein levels were decreased in brainstem at the age of 6 months. These results suggest the presence of a chronic neuroinflammation process with deficiency of remyelination-stimulating factors (CXCL1, CXCR2, and CXCR4), which might account for the demyelination in thetaieprat.

scholarly journals Resolution and limitations of the immunoperoxidase procedure in the localization of extracellular matrix antigens.

1983 ◽  
Vol 31 (7) ◽  
pp. 945-951 ◽  
Author(s):  
P J Courtoy ◽  
D H Picton ◽  
M G Farquhar
Keyword(s):  
Extracellular Matrix ◽  
Basement Membrane ◽  
Glomerular Basement Membrane ◽  
Cationized Ferritin ◽  
Anionic Sites ◽  
Double Labeling ◽  
Extracellular Matrix Components ◽  
Matrix Components ◽  
Indirect Immunoperoxidase ◽  
Labeling System

A double labeling system was used to test the resolution of the indirect immunoperoxidase procedure in the localization of extracellular matrix components. A recognizable antigen, cationized ferritin, was first implanted at specific anionic sites (approximately 60 nm periodicity) in the lamina rara interna and externa of the glomerular basement membrane (GBM) and subsequently localized by immunoperoxidase. The coincidence between the location of reaction product and the ferritin clusters was assessed. When the amount of immunoadsorbed peroxidase and time of exposure to the 3,3'-diaminobenzidine (DAB)-containing medium were limited, discrete deposits of reaction product were observed around individual ferritin clusters. When immunolabeling was increased, the whole GBM was stained, and DAB staining was also found along the endothelial plasmalemma and the epithelial plasmalemma at the base of the foot processes at some distance (greater than 100 nm) from the ferritin clusters in the laminae rarae. These findings indicate that oxidized DAB reaction product can diffuse over long distances and be reabsorbed onto cell membranes. Even under limited incubation conditions some diffusion of DAB reaction product was encountered. The value and limitations of the DAB-peroxidase procedures are discussed.

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