Enhanced Hexachloro-1:3-butadiene Nephrotoxicity in Rats with a Preexisting Adriamycin-Induced Nephrotic Syndrome

Abstract

Renal damage was assessed by histopathology and urinalysis in male Wistar rats treated with either hexachloro-1:3-butadiene (HCBD; a single 170-mg/kg ip dose that caused proximal tubule necrosis), adriamycin (ADR; a single 5-mg/kg ip dose that caused minimal glomerular changes up to 35 days), or HCBD given 2 wk after ADR and compared with age-matched control rats for 21 days. Urinalysis values in ADRtreated rats showed minimal renal changes. HCBD significantly elevated urine volume (10-fold), protein (5-fold), glucose (175-fold), and brush border enzymes (10-600-fold), indicating severe proximal tubular damage, but most parameters returned to pretreatment levels 6 days after treatment. In ADR-pretreated rats subsequently given HCBD, both the urinary alkaline phosphatase and the ratio of kidney : body weight were significantly higher for longer periods. Histopathology demonstrated that the HCBD-induced proximal tubular lesion was confined to the outer stripe of the outer medulla. Advanced regeneration and repair was evident 21 days after HCBD treatment. In the ADR-pretreated rats the HCBD-induced lesion was more severe and affected the entire cortex (24).Statistics. Statistically significant differences between groups were determined by 2-tailed Students' t-test, with p < 0.05 considered to be statistically significant. RESULTS UrinanalysisThere were no significant differences in the urinary parameters in the control and ADR groups ( (Fig. 1 a). Tubules were dilated and filled with homogenous eosinophilic material and necrotic epithelial cells. Signs of regeneration and restitution (i.e., tubular re-epithelialization and loss of tubular casts and debris) were evident by day 7 and complete 21 days after treatment (Fig. 1 b).Histologically, animals in the ADR/HCBD group showed much more extensive renal damage (Fig. 2a) when compared to those treated with HCBD alone (see Fig. 1 a). Extensive tubular damage was present throughout the entire cortex 2 days after HCBD treatment, and tubules were dilated and filled with homogeneous proteinaceous material. Sloughed necrotic calcified epithelial cells were observed in tubular lumens and casts extended deep into the medulla (Fig. 2b). Extensive calcification was evident by day 7 (which was not seen in the HCBD only group) and by day 21, regeneration, although apparent, was less advanced and marked tubular calcification was still evident (Fig. 2c). Table 2 summarizes the differences between the ADR/HCBD group and HCBD alone. HistochemistryControl kidneys demonstrated normal GGT staining of the tubular epithelium of the outer stripe of the outer medulla (Fig. 3a). ADR caused no changes in this staining pattern. GGT staining in the outer stripe of the outer medulla was diminished 24 hr after HCBD treatment, was barely evident by day 3 (Fig. 3b) HCBD is a useful model toxin for damaging the proximal tubule. It is metabolized in the liver, ex-

Urinary biomarkers in hexachloro‐1:3‐butadiene‐induced acute kidney injury in the female Hanover Wistar rat; correlation of α‐glutathione S‐transferase, albumin and kidney injury molecule‐1 with histopathology and gene expression

“…). This finding is consistent with previous observations of recovery by day 14 after administration of 200 mg kg −1 HCBD (Ishmael et al , ) and evidence of advanced regeneration and repair at 21 days after 170 mg kg −1 HCBD administration (Kirby and Bach, ).…”

Correlation of Histopathology, Urinary Biomarkers, and Gene Expression Responses Following Hexachloro-1:3-Butadiene–induced Acute Nephrotoxicity in Male Hanover Wistar Rats

“…This was attributed to the diuresis effect of glycosuria (Table 2). Increased concentrations of urinary glucose have been associated with acute tubular necrosis and have been reported previously in HCBD-treated rats (Kirby and Bach 1995; Swain et al 2011, 2012).…”

“…This work has demonstrated the sensitivity of the urinary biomarkers in detecting early renal tubular injury in response to increasing dose levels of HCBD. However, as the kidney undergoes an initial phase of acute degeneration after HCBD administration, followed by a relatively rapid return to normal morphology (Kirby and Bach 1995), the temporal effect of HCBD administration on the more recent renal biomarkers can also be characterized using this animal model. Urinary output of renal biomarkers that are closely aligned to cytoplasmic leakage, such as the urinary glutathione S-transferases (GSTs; Harrison et al 1989), may differ from the production of biomarkers associated with functional and morphologic recovery.…”

Effects of long-term oral treatment with selective vasopressin V2 receptor antagonist (OPC-31260) on adriamycin-induced heart failure in rats

“…Single intravenous injections of adriamycin (as low as 5 mg/kg) into rats have been used to induce experimental nephrotic syndrome characterized by heavy proteinuria, hypoalbuminemia, hypercholesterolemia, thoracic and ascitic fluid [26]. However, it was reported that single intraperitoneal injections of adriamycin (5 mg/kg) into rats induced minimal renal changes [27]. In our study, we also used the intraperitoneal adriamycin, and we might think renal damage was minor in the animal model used in this study and the damage did not interfere with the study for a CHF model.…”