Elsevier

Biochemical Pharmacology

Volume 64, Issue 3, 1 August 2002, Pages 517-525
Biochemical Pharmacology

Effect of pirfenidone against vanadate-induced kidney fibrosis in rats

https://doi.org/10.1016/S0006-2952(02)01213-3Get rights and content

Abstract

Renal fibrosis is a complication of kidney injury and can contribute to organ failure. Currently, there are no drugs for the treatment of renal fibrosis. Pirfenidone (PD) has been proven to have antifibrotic effects in animal models of fibrosis. We tested the ability of PD against vanadate-induced kidney fibrosis in rats. The rats were injected subcutaneously with vehicle or vanadate solution (1 mg vanadate/kg/day) for 12 or 16 days to produce varying degrees of kidney fibrosis. The vanadate- and vehicle-treated rats were fed a laboratory diet or the same diet mixed with 0.6% PD ad lib. One vanadate-injected group was initially fed the same diet without PD and later switched to the diet containing PD 2 days after the last injection. The rats were killed at 12 and 25 days following the last dose. The changes found in the kidney of vanadate-treated rats included increases in RNA and DNA content and increases in kidney weight. Treatment with PD diminished the vanadate-induced increases in kidney weight and RNA content. The hydroxyproline content of the kidney in vanadate-treated animals was increased significantly (P≤0.05) from the control level of 1452 μg/kidney to 1765 μg/kidney. Treatment with PD for 37 days caused significant reductions in the vanadate-induced increases in the hydroxyproline level. Similarly, treatment for 41 days also caused significant reductions (1744 μg/kidney) in vanadate-induced increases in the hydroxyproline level (1996 μg/kidney). The histological evaluation revealed that the severity of the lesions in the vanadate-treated group was moderate to severe, and treatment with PD for 41 days decreased the severity to a mild level. In addition, the delayed treatment with PD also minimized the vanadate-induced increases in the collagen content of the kidney. Although it is speculative, PD may potentially be therapeutic in the management of renal fibrosis.

Introduction

Kidney fibrosis may develop as a result of chronic infection, obstruction of the ureter by calculi, malignant hypertension, severe diabetic conditions, or chronic exposure to heavy metals such as lead, vanadium, and mercury [1], [2], [3], [4]. In addition, idiopathic glomerulosclerosis and renal interstitial fibrosis have been reported in children and adults [5], [6]. Kidney fibrosis correlates well with the overall loss of renal function and could contribute to renal failure [7]. At the present time, fibrotic diseases of various organs in humans are treated chronically with high doses of corticosteriods and cytotoxic drugs that have serious systemic adverse effects [8], [9].

The antifibrotic effect of PD has been demonstrated in several animal models of fibrosis in different organs. For example, the dietary intake of PD ameliorates bleomycin- and cyclophosphamide-induced lung fibrosis in hamsters and mice [10], [11], [12], [13]. Dietary intake of PD reduces collagen accumulation in the remnant kidney with partial nephrectomy and post-obstructive models of kidney fibrosis in rats [14], [15]. In addition, treatment with PD and spirnolactone reverses cardiac and renal fibrosis in streptozotocin-diabetic rats [16]. Interestingly, Raghu et al. [17] demonstrated the beneficial effects of PD against idiopathic pulmonary fibrosis in an open clinical trial in humans with advanced and end-stage fibrosis.

In humans, loss of kidney functions due to fibrosis usually occurs many years after the onset of diabetes or recurrent infections. The use of animal models for kidney fibrosis in the evaluation of drugs for their potential antifibrotic effects may not only help to elucidate the mechanisms of pathogenesis of renal fibrosis, but may also lead to the development of efficacious drugs for the treatment of this debilitating disease in humans. At present, animal models of kidney fibrosis include: (a) ligation of the ureter and partial nephrectomy [14], [15], and (b) chronic treatment with cyclosporin A (CsA) in rats on a low-salt diet [18]. The ligation and partial nephrectomy models of kidney fibrosis may alter the systemic and renal physiological processes of the animals, and the CsA model leads to fibrotic changes confined mostly to the medullary region of the kidney [18]. We have demonstrated in a previous study that subcutaneous injections of vanadate in rats at a daily dose of 0.6–0.9 mg/kg for 16 days cause inflammation, cellular hyperplasia, and fibrosis of both the cortical and medullary regions of the kidney when examined at different times during the 25 days following the last injection [1]. These changes appeared to be similar to those found in kidneys of humans at acute, subacute, and chronic stages of inflammation. Consequently, we employed the vanadate-rat model of kidney fibrosis to determine if PD is effective in ameliorating kidney fibrosis. The data presented in this paper indicate that dietary intake of PD prevented the development of vanadate-induced kidney fibrosis, and it also arrested the ongoing and progressive fibroproliferative process of the kidney. It was concluded that PD could be an effective drug for the management of kidney fibrosis resulting from various medical conditions in humans.

Section snippets

Animals and treatment

A total of 126 male Sprague–Dawley rats (Charles River), 9 weeks old, were used in this study. The average initial weight of these animals was 351±1 g (±SEM). The rats were housed individually in polycarbonate cages in facilities approved by the American Association for the Accreditation of Laboratory Animal Care. These animals were acclimatized to laboratory conditions for 1 week prior to the start of the experiment. A 12 hr/12 hr light/dark cycle was maintained in the housing facilities. The

Results

Since there were no differences in any of the measured biochemical determinants between the two vehicle-injected control groups (one on the RD; and the other on the same diet containing PD), the data from both groups were pooled and depicted as the RD control. The levels of hydroxyproline in kidneys of vanadate-treated rats in both Study No. 1 and Study No. 2 were significantly higher in the RD+V groups than the levels in the corresponding vehicle-treated rats in the control RD groups (see Fig.

Discussion

Data presented in this study revealed that vanadate treatment affected both cortical and medullary regions of the kidney and produced moderate to severe damage leading to fibrosis of both regions. The acute stage of inflammation was followed by cortical and medullary fibrosis and medullary epithelial cell hyperplasia. The severity of the lesions and fibrosis was dependent upon the cumulative amount of injected vanadate. The histological and biochemical changes observed in kidneys were similar

Acknowledgements

The authors would like to thank Marnac, Inc. for financial support and for the supply of PD for the study.

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