Heparin Treatment Reduces Glomerular Injury in Rats with Adriamycin-Induced Nephropathy but Does Not Modify Tubulointerstitial Damage or the Renal Production of Transforming Growth Factor-Beta

Abstract

In this study we investigated the effect of heparin on renal injury and renal transforming growth factor-β (TGF-β) production in adriamycin (AD)-injected rats. Thirty-nine female Wistar rats were injected with AD (3.5 mg/kg body weight, i.v.) and 27 with 0.15 M NaCl solution (group C). Fifteen days later we started to inject heparin, 500 U/day, s.c., in 20 of the AD-injected animals (AD-H group). Three months after beginning treatment, urine samples were collected to quantify albumin, creatinine and TGF-β. The rats were killed and the kidneys removed for histological, immunohistochemical, ELISA and RNA studies. All AD-injected animals showed structural renal changes (p < 0.05). However, the glomerular alterations were less intense in rats from group AD-H (p < 0.05). The percentage of glomerulosclerosis was 0.11 ± 0.08 in group C, 14.7 ± 12.8 in group AD (treated only with AD) and 3.42 ± 2.3 in group AD-H. Renal cortex immunostaining for TGF-β and mRNA content of this polypeptide was higher in both groups of animals injected with AD compared to controls (p < 0.05). These animals also presented a higher rate of urinary TGF-β excretion (p < 0.05), which was 202 ± 11 in group C, 1,103 ± 580 in group AD and 1,564 ± 328 pg/mg Ucreat in group AD-H. However, TGF-β activity in the glomerular-conditioned media from the rats of group AD was higher than in the glomerular-conditioned media from the rats of group AD-H. In conclusion, treatment with heparin reduces glomerular damage in rats with AD-induced nephropathy but does not modify tubulointerstitial lesions or the renal production of TGF-β.

Inhibition of Nuclear Factor-κB Activation Attenuates Tubulointerstitial Nephritis Induced by Gentamicin

“…4-µm histological sections were stained with Masson’s trichrome and examined under the light microscope. The fractional interstitial area of the renal cortex was determined by morphometry with a light camera connected to an image analyzer (Kontron Electronic System KS 300, Eching, Germany) [14]. Twenty grid fields measuring 0.174 mm 2 were evaluated in the renal cortex section of each kidney.…”

“…The kidneys from 7 control animals, 14 from animals killed 5 days after gentamicin treatment and 14 from rats killed 30 days after treatment were subjected to immunohistochemical staining [14, 16]. …”

“…For evaluation of immunoperoxidase staining for NF-ĸB, pERK and pJNK, each tubulointerstitial grid field was graded semiquantitatively, and the mean score per kidney was calculated [14]. Each score reflected mainly changes in the extent, rather than the intensity, of staining and depended on the percentage of grid field showing positive staining: 0, absent or <5%; I, 5–25%; II, 25–50%; III, 50–75%, and IV, >75%.…”

Long‐term evolution of the acute tubular necrosis (ATN) induced by glycerol: role of myofibroblasts and macrophages

“…Encircled damaged areas were traced manually on a video screen, and determined by computerized morphometry (Baroni et al 2000).…”

“…The rats were subjected to aortic perfusion with PBS (0.15 M NaCl and 0.01 phosphate buffer, pH 7.4) until the kidneys were blanched. The kidneys were then perfused with 4% paraformaldehyde, fixed in paraformaldehyde 4% for 2 h, postfixed in Bouin's solution for an additional 4±6 h, rinsed with 70% ethanol to eliminate picric acid, dehydrated through a graded alcohol series, embedded in paraffin, sectioned into 3-mm slices, deparaffinized, and subjected to immunohistochemical staining (Kliem et al 1996;Baroni et al 2000). The sections were incubated overnight at 4 C with 1/500 antifibronectin and 1/30 anti-TGF-b polyclonal antibodies produced in rabbits or 1/1000 anti a-SM-actin (overnight) and 1/2500 anti-ED-1 (1 h) monoclonals antibodies.…”