Transplantation/ImmunologyThe Experimental Agent Pirfenidone Reduces Pro-Fibrotic Gene Expression in a Model of Tacrolimus-Induced Nephrotoxicity
Introduction
The use of calcineurin-inhibitor based immunosuppressants is a contributory factor in chronic renal allograft dysfunction (CRAD), the pathophysiological entity that is the leading cause of allograft loss after the first post-transplant year [1]. Evidence for the nephrotoxicity of cyclosporine is now well established [2], and a number of short- and long-term animal and human studies in the setting of organ allografting have demonstrated the nephrotoxic potential of tacrolimus [3, 4, 5]. The histopathological changes in renal allograft biopsies, consisting of tubular vacuolisation, striped interstitial fibrosis and arteriolar hyalinosis, are associated with both long-term cyclosporine and tacrolimus therapy [6].
The rat salt-depleted model [7] has become a paradigm for the study of chronic calcineurin-inhibitor nephrotoxicity, because it allows examination of the underlying mechanisms without the influence of multiple confounding factors that can confuse the findings of clinical studies. It has elucidated some of the structural, functional [5, 8] and molecular mechanisms [3] of tacrolimus nephrotoxicity, which include alterations in the levels of profibrotic cytokines and changes in extracellular matrix metabolism.
To date, no pharmacological agents have proved effective in the clinical setting of CRAD. Pirfenidone (5-methyl-1-phenyl-2-(1H)-pyridone; Marnac Inc., Dallas, TX) is a novel anti-fibrotic agent that has demonstrated potential in a number of animal models of fibrotic disease. It can prevent or reverse ECM accumulation in several organs [9, 10, 11], by decreasing the expression of profibrotic cytokines such as TNF-α and TGF-β. The aim of this study was to examine the effects of pirfenidone on physiological and histological markers of tacrolimus-induced renal injury in the rat salt-depleted model. Further, we appraised the underlying molecular mechanisms, looking specifically at the mRNA expression of genes involved in extracellular matrix turnover and fibrosis. We hypothesized that pirfenidone reduces the renal functional, histological, and molecular markers of tacrolimus-induced fibrosis.
Section snippets
Treatment schedule
Male Sprague-Dawley rats (350–500 g), obtained from Harlan (Cambridge, U.K.) were housed and cared for in accordance with the Animals (Scientific Procedures) Act 1986, in cages of three animals in a temperature and light controlled environment, with water ad libitum. The rats were acclimatized for 7 days on a 12:12 h light:dark cycle. They were fed a salt-depleted diet for 7 days (0.05% sodium, Special Diets Services, Witham, Essex, U.K.), before the introduction of a regimen of tacrolimus
Animal weight
Animals in all groups gained weight at an approximately equal rate, and there were no significant differences in weight across groups (data not shown). This suggests that neither tacrolimus nor pirfenidone treatment displayed systemic toxicity.
Serum creatinine
Tacrolimus treatment caused a rise in serum creatinine compared to the low salt-diet treated controls at 21 days (75 versus 55 μmol/l, P = 0.001) and 28 days (76 versus 61 μmol/l, P = 0.01) (see Table 2). Treatment with tacrolimus plus pirfenidone (at all
Discussion
This study has demonstrated that pirfenidone attenuates the creatinine rise seen in a model of chronic tacrolimus toxicity. It has also shown a reduction in both TGF-β and TIMP-1 expression by tacrolimus. Although pirfenidone had no effect on TGF-β, TIMP-1 levels were further reduced by its addition, and collagen III expression was reduced below control levels by high-dose pirfenidone. Neither tacrolimus nor pirfenidone altered the expression of the matrix metalloproteinases 2 and 9. Renal
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Pirfenidone is a renal protective drug: Mechanisms, signalling pathways, and preclinical evidence
2021, European Journal of PharmacologyCitation Excerpt :Further, Shihab et al. noted that PFD could suppress the expression of pro-apoptotic genes (p53 and fas-ligand) and down-regulate caspase 3 expression in the CsA-induced renal fibrosis model (Shihab et al., 2005), which demonstrates the anti-apoptotic ability of PFD. PFD has been evaluated in a variety of experimental models of primary and secondary kidney diseases, including 5/6 nephrectomised rats (Takakuta et al., 2010; Chen et al., 2013a; Shimizu et al., 1997; Chen et al., 2013b), FGS/Kist mouse (Park et al., 2003), streptozotocin-diabetic rats (Miric et al., 2001), db/db mice (RamachandraRao et al., 2009), hypertension-induced renal injury (Ji et al., 2013), UUO (Shimizu et al., 1998; Li et al., 2017), acute kidney injury due to bilateral renal ischemia (Lima-Posada et al., 2019), chronic renal allograft dysfunction (Qiu et al., 2019), and drug-induced renal fibrosis (Shihab et al., 2002, 2005; Brook et al., 2005; Al-Bayati et al., 2002; Sharawy and Serrya, 2020) (Table 1). Mechanical stress, oxidative stress, inflammation, and metabolic and vasoactive factors (such as high glucose level and activation of the renin-angiotensin-aldosterone system) can increase ROS production, mitochondrial damage and TGF-β expression, and impair kidney function in glomerular-related diseases, which may be alleviated by PFD (Fig. 1).
Antiadhesive and anti-inflammatory effects of pirfenidone in postoperative intra-abdominal adhesion in an experimental rat model
2016, Journal of Surgical ResearchCitation Excerpt :Clinical studies in idiopathic pulmonary fibrosis have demonstrated the effectiveness of PF in extending survival time and improving pulmonary function [13–15]. PF has also been shown to have beneficial effects in renal and liver fibrosis, and it exerts those effects by upregulating the transcription of matrix metalloproteinases (MMPs) and downregulating the transcription of tissue inhibitors of metalloproteinases (TIMPs) [16,17]. PF is highly soluble in aqueous solutions and is able to move through cell membranes without requiring a receptor.
Aristolochic acid downregulates monocytic matrix metalloproteinase-9 by inhibiting nuclear factor-κB activation
2011, Chemico-Biological InteractionsCitation Excerpt :On the other hand, it was proposed that a decrease in TIMP-1 levels could downregulate the inhibitory influences acting on MMPs, causing a higher degree of matrix degradation. Therefore, lowered TIMP-1 levels are thought to abrogate renal fibrosis [22,50]. Through marked infiltration of monocytes/macrophages in renal lesions, the expression of TIMP-1 was lower in such leukocytes during the fibrotic process of chronic cyclosporine nephropathy [51] and unilateral ureteral obstruction [52].
Pirfenidone: A Novel Potential Therapeutic Agent in the Management of Chronic Allograft Rejection
2007, Transplantation ProceedingsCitation Excerpt :The results indicated that in the rats receiving pirfenidone, profibrotic genes were downregulated. These genes included TGF-β, TIMP-1, and collagen III, and there was less matrix deposition in the renal cortex.26–28 Further investigation may be warranted to elucidate the exact mechanisms of the drug interaction.
Pirfenidone: Anti-fibrotic agent with a potential therapeutic role in the management of transplantation patients
2006, European Journal of Pharmacology
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Nicholas Brook is partly funded by The Royal College of Surgeons of Edinburgh Robertson Trust Fellowship.