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Nephroprotective effect of heparanase in experimental nephrotic syndrome.

Assady S, Alter J, Axelman E, Zohar Y, Sabo E, Litvak M, Kaplan M, Ilan N, Vlodavsky I, Abassi Z - PLoS ONE (2015)

Bottom Line: Heparanase, an endoglycosidase that cleaves heparan sulfate (HS), is involved in various biologic processes.Albuminuria was investigated at days 0, 7, and 14 thereafter.Our results suggest that heparanase may play a nephroprotective role in ADR-NS, most likely independently of HS degradation.

View Article: PubMed Central - PubMed

Affiliation: Department of Nephrology, Rambam Health Care Campus, Haifa, Israel.

ABSTRACT

Background: Heparanase, an endoglycosidase that cleaves heparan sulfate (HS), is involved in various biologic processes. Recently, an association between heparanase and glomerular injury was suggested. The present study examines the involvement of heparanase in the pathogenesis of Adriamycin-induced nephrotic syndrome (ADR-NS) in a mouse model.

Methods: BALB/c wild-type (wt) mice and heparanase overexpressing transgenic mice (hpa-TG) were tail-vein injected with either Adriamycin (ADR, 10 mg/kg) or vehicle. Albuminuria was investigated at days 0, 7, and 14 thereafter. Mice were sacrificed at day 15, and kidneys were harvested for various analyses: structure and ultrastructure alterations, podocyte proteins expression, and heparanase enzymatic activity.

Results: ADR-injected wt mice developed severe albuminuria, while ADR-hpa-TG mice showed only a mild elevation in urinary albumin excretion. In parallel, light microscopy of stained cross sections of kidneys from ADR-injected wt mice, but not hpa-TG mice, showed mild to severe glomerular and tubular damage. Western blot and immunofluorescence analyses revealed significant reduction in nephrin and podocin protein expression in ADR-wt mice, but not in ADR-hpa-TG mice. These results were substantiated by electron-microscopy findings showing massive foot process effacement in injected ADR-wt mice, in contrast to largely preserved integrity of podocyte architecture in ADR-hpa-TG mice.

Conclusions: Our results suggest that heparanase may play a nephroprotective role in ADR-NS, most likely independently of HS degradation. Moreover, hpa-TG mice comprise an invaluable in vivo platform to investigate the interplay between heparanase and glomerular injury.

No MeSH data available.


Related in: MedlinePlus

Heparanase protects podocytes from Adriamycin induced injury.wt or hpa-TG mice were injected with Adriamycin (ADR) or served as control (sham). Two weeks post injection, the animals were sacrificed. (A) Representative images of transmission electron microscopy of ultrathin sections of kidney tissue. Magnification X15 000; scale bar = 1μm. (B) Quantification of foot process width (n = 12 glomeruli per each wt group, 13 for hpa-TG, and 11 for hpa-TG ADR group, obtained from 6, 3, 4, and 5 animals, respectively). *, P<0.01 vs. all other groups.
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pone.0119610.g004: Heparanase protects podocytes from Adriamycin induced injury.wt or hpa-TG mice were injected with Adriamycin (ADR) or served as control (sham). Two weeks post injection, the animals were sacrificed. (A) Representative images of transmission electron microscopy of ultrathin sections of kidney tissue. Magnification X15 000; scale bar = 1μm. (B) Quantification of foot process width (n = 12 glomeruli per each wt group, 13 for hpa-TG, and 11 for hpa-TG ADR group, obtained from 6, 3, 4, and 5 animals, respectively). *, P<0.01 vs. all other groups.

Mentions: This approach, though a hallmark characteristic of glomerular injury, does not necessarily indicate podocytes’ fate. Hence, electron microscopy was applied. In line with the observed massive proteinuria, Adriamycin-injected wild type mice demonstrated flattening and effacement of foot processes (Fig. 4). While, analysis of ultrathin kidney sections of injected hpa-TG mice revealed normal podocyte architecture with numerous, intact foot processes and slit diaphragm (Fig. 4). Mean foot process width was significantly increased (1470 ± 244 nm) in ADR-injected wt mice vs. wt controls, ADR-injected and control hpa-TG mice (593 ± 60 nm, 599 ± 100 nm, and 600 ± 41 nm, respectively, Fig. 4B). Furthermore, in pilot experiments where PEI staining was used, we demonstrated that hpa-TG mice displayed a decrease in the GBM anionic charge as compared with wt controls. In addition, disruption of these anionic sites was observed in both ADR-injected wt and hpa-TG mice (S2 Fig.). Such a reduction in negative charge sites did not correlate with the degree of albuminuria presented in Fig. 1A, suggesting a distinctive role of heparanase not related to charge permselectivity or HSPGs.


Nephroprotective effect of heparanase in experimental nephrotic syndrome.

Assady S, Alter J, Axelman E, Zohar Y, Sabo E, Litvak M, Kaplan M, Ilan N, Vlodavsky I, Abassi Z - PLoS ONE (2015)

Heparanase protects podocytes from Adriamycin induced injury.wt or hpa-TG mice were injected with Adriamycin (ADR) or served as control (sham). Two weeks post injection, the animals were sacrificed. (A) Representative images of transmission electron microscopy of ultrathin sections of kidney tissue. Magnification X15 000; scale bar = 1μm. (B) Quantification of foot process width (n = 12 glomeruli per each wt group, 13 for hpa-TG, and 11 for hpa-TG ADR group, obtained from 6, 3, 4, and 5 animals, respectively). *, P<0.01 vs. all other groups.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4364762&req=5

pone.0119610.g004: Heparanase protects podocytes from Adriamycin induced injury.wt or hpa-TG mice were injected with Adriamycin (ADR) or served as control (sham). Two weeks post injection, the animals were sacrificed. (A) Representative images of transmission electron microscopy of ultrathin sections of kidney tissue. Magnification X15 000; scale bar = 1μm. (B) Quantification of foot process width (n = 12 glomeruli per each wt group, 13 for hpa-TG, and 11 for hpa-TG ADR group, obtained from 6, 3, 4, and 5 animals, respectively). *, P<0.01 vs. all other groups.
Mentions: This approach, though a hallmark characteristic of glomerular injury, does not necessarily indicate podocytes’ fate. Hence, electron microscopy was applied. In line with the observed massive proteinuria, Adriamycin-injected wild type mice demonstrated flattening and effacement of foot processes (Fig. 4). While, analysis of ultrathin kidney sections of injected hpa-TG mice revealed normal podocyte architecture with numerous, intact foot processes and slit diaphragm (Fig. 4). Mean foot process width was significantly increased (1470 ± 244 nm) in ADR-injected wt mice vs. wt controls, ADR-injected and control hpa-TG mice (593 ± 60 nm, 599 ± 100 nm, and 600 ± 41 nm, respectively, Fig. 4B). Furthermore, in pilot experiments where PEI staining was used, we demonstrated that hpa-TG mice displayed a decrease in the GBM anionic charge as compared with wt controls. In addition, disruption of these anionic sites was observed in both ADR-injected wt and hpa-TG mice (S2 Fig.). Such a reduction in negative charge sites did not correlate with the degree of albuminuria presented in Fig. 1A, suggesting a distinctive role of heparanase not related to charge permselectivity or HSPGs.

Bottom Line: Heparanase, an endoglycosidase that cleaves heparan sulfate (HS), is involved in various biologic processes.Albuminuria was investigated at days 0, 7, and 14 thereafter.Our results suggest that heparanase may play a nephroprotective role in ADR-NS, most likely independently of HS degradation.

View Article: PubMed Central - PubMed

Affiliation: Department of Nephrology, Rambam Health Care Campus, Haifa, Israel.

ABSTRACT

Background: Heparanase, an endoglycosidase that cleaves heparan sulfate (HS), is involved in various biologic processes. Recently, an association between heparanase and glomerular injury was suggested. The present study examines the involvement of heparanase in the pathogenesis of Adriamycin-induced nephrotic syndrome (ADR-NS) in a mouse model.

Methods: BALB/c wild-type (wt) mice and heparanase overexpressing transgenic mice (hpa-TG) were tail-vein injected with either Adriamycin (ADR, 10 mg/kg) or vehicle. Albuminuria was investigated at days 0, 7, and 14 thereafter. Mice were sacrificed at day 15, and kidneys were harvested for various analyses: structure and ultrastructure alterations, podocyte proteins expression, and heparanase enzymatic activity.

Results: ADR-injected wt mice developed severe albuminuria, while ADR-hpa-TG mice showed only a mild elevation in urinary albumin excretion. In parallel, light microscopy of stained cross sections of kidneys from ADR-injected wt mice, but not hpa-TG mice, showed mild to severe glomerular and tubular damage. Western blot and immunofluorescence analyses revealed significant reduction in nephrin and podocin protein expression in ADR-wt mice, but not in ADR-hpa-TG mice. These results were substantiated by electron-microscopy findings showing massive foot process effacement in injected ADR-wt mice, in contrast to largely preserved integrity of podocyte architecture in ADR-hpa-TG mice.

Conclusions: Our results suggest that heparanase may play a nephroprotective role in ADR-NS, most likely independently of HS degradation. Moreover, hpa-TG mice comprise an invaluable in vivo platform to investigate the interplay between heparanase and glomerular injury.

No MeSH data available.


Related in: MedlinePlus