Regulation of renal organic anion and cation transporters by thymoquinone in cisplatin induced kidney injury

https://doi.org/10.1016/j.fct.2012.02.082Get rights and content

Abstract

In previous studies, we have demonstrated the biological activity of thymoquinone (TQ), an active compound extracted from the Nigella sativa plant, against cisplatin-induced neurotoxicity. Recenty, it was observed that there is an inherent lack in regulation of renal organic anion and cation transporters in cisplatin-induced nephrotoxicity. Here, we report, for the first time, the effect of TQ on alterations in the renal expression of organic anion transporters (OATs) and organic cation transporters (OCTs), as well as multidrug resistance-associated proteins (MRPs) in rats treated with cisplatin. Twenty-eight 8-week-old male Wistar rats were divided into four groups of control, TQ treated (10 mg/kg b.w. in drinking water for 5 days), cisplatin (7 mg/kg b.w., i.p.) and TQ and cisplatin combination treatment. Cisplatin-induced malondialdehyde (MDA) and 8-isoprostane increase was found to be markedly reduced in rats treated with TQ. In cisplatin only treated rats, the induced renal injury increased protein levels of the efflux transporters MRP2 and MRP4 while expression of OAT1, OAT3, OCT1 and OCT2 was reduced. In combination TQ- and cisplatin-treated rats, expression of MRP2 and MRP4 proteins was decreased in the kidneys. Conversely, TQ treatment increased levels of OCT1, OCT2, OAT1 and OAT3 and decreased levels of 8-isoprostane and MDA levels in cisplatin-treated rats. In conclusion, the present study shows that the TQ synergizes with its nephroprotective effect against cisplatin-induced acute kidney injury in rats.

Highlights

Cisplatin causes nephrotoxicity. ► Thymoquinone (TQ) is the bioactive compound derived from Nigella sativa. ► TQ increased levels of OCTs and OATs in cisplatin-treated rats. ► TQ decreased levels of MRP2, MRP4 in cisplatin-treated rats.

Introduction

Cisplatin, one of the most effective and potent anticancer drugs, is used in the treatment of a wide variety of both pediatric and adult malignancies (Ramesh and Reeves, 2002). However, the chemotherapeutic use of cisplatin is limited by serious side-effects such as nephrotoxicity and ototoxicity, sometimes requiring a reduction in dose or discontinuation of treatment. Even though nephrotoxicity can be managed with some success by concomitant use of intravenous hydration, it is still a major factor that limits administration and efficacy of cisplatin in cancer therapy (Pabla and Dong, 2008). Cisplatin chemotherapy induces a reduction in antioxidant status, leading to a failure of the antioxidant defense against free-radical damage generated by antitumor drugs (Arjumand et al., 2011, Masuda et al., 1994). It has been previously shown that cisplatin-induced-nephrotoxicity is closely associated with an increase in lipid peroxidation in the kidney tissues as well (Antunes et al., 2000).

TQ is the bioactive compound derived from black seed (Nigella sativa) oil. TQ has been shown to exert anti-inflammatory, anti-oxidant and anti-neoplastic effects both in vitro and in vivo. Several studies reported that TQ showed significant anti-neoplastic activity against human pancreatic adenocarcinoma, uterine sarcoma and leukemic cell lines, while it is minimally toxic to normal cells (Worthen et al., 1998). In addition, orally administered TQ potentiated cisplatin antitumor activity and prevented cisplatin-induced nephrotoxicity in mice and rats as evidenced by significant reductions in serum urea and significant improvement in kidney weight and creatinine clearance (Badary et al., 1997).

The kidney plays an important role in the elimination of numerous endogenous and exogenous organic anions and cations, including efflux various drugs and endogenous metabolites from blood into the urine (Morisaki et al., 2008). Transporter-mediated secretory pathways for organic anions and cations exist that facilitate active secretion of a wide range of exogenous and endogenous organic ions in the renal proximal tubules (Morisaki et al., 2008, Pritchard and Miller, 1996). Renal basolateral isoforms of multispecific organic anion and cation transporters OAT1, OAT3, OCT1 and OCT2 have been cloned and characterized (Morisaki et al., 2008). In experimental nephrotoxicity, a significant reduction in tubular cation clearance was observed. This was directly associated with a down-regulation of the expression of tubular organic anion and cation transporters (OCTs) (Huang et al., 2001). To protect against such harmful metabolites, biological system have developed certain clearance mechanisms that involved cation transporters such as OCTs and OATs. OCTs and OATs are necessary for the renal clearance of a broad range of exogenous substrates, including toxins, xenobiotics, and commonly used drugs. Additionally, the MRP subfamily of ABC transporters (MRP/ABCC) are membrane glycoproteins that mediate the ATP-dependent export of organic anions, including cytotoxic and antiviral drugs, from cells.

Altered expression of renal OATs and OCTs and MRPs has been reported in animal models with renal impairment induced by cisplatin (Aleksunes et al., 2008, Ciarimboli et al., 2005, Ji et al., 2002, Laouari et al., 2001). Administration of a single dose of cisplatin to rats reduces OCT2 expression at 7 days, suggesting a defense response against subsequent exposure and renal uptake of cisplatin (Huang et al., 2001). Cisplatin treatment also increases expression of renal MRP2 in rats (Demeule et al., 1999, Huang et al., 2001). However, there is no information on the protective effect of TQ on the alterations in the expression of renal OATs and OCTs and MRPs during cisplatin-induced renal injury of rats. Therefore, the purpose of this study was to investigate the effects of TQ on expression of OATs, OCTs and MRPs in kidneys in cisplatin-treated rats.

Section snippets

Animals

Eight-week-old male Wistar rats (180–245 g) were obtained from Firat University Research Center (Elazig, Turkey). The rats were were housed in a ventilated room at 25 ± 5 °C under a 12-h light/dark cycle. The animals were acclimatized for 1 week before the study and had free access to standard laboratory feed and water ad libitum. This study was reviewed by the Committee for Ethics in Animal Experiments of the Firat University and carried out under Turkish law and the Guidelines for Animal

Effect of TQ on cisplatin-induced oxidative stress markers

First we evaluated serum urea and creatinine concentrations that were found to be significantly increased in cisplatin-treated rats (cisplatin, 152.1 ± 10.5 and 1.46 ± 0.18 vs. normal control, 53.5 ± 15.8 and 0.43 ± 0.08, respectively; P < 0.05) (Table 1). However, a marked decrease in serum urea and creatinine were observed upon administration of TQ in cisplatin-treated rats. Table 2 summarizes the alternations in the indicators of oxidative stress, namely MDA and 8-isoprostane levels, in kidney tissues

Discussion

The presented findings demonstrate that TQ can reverse cisplatin-stimulated oxidative stress makers, lipid peroxidation status and other nephrotoxicity markers. These studies build a strong case for the use of TQ as a protective agent in cisplatin-based chemotherapy regimens.

Cisplatin stimulates renal production of oxidative stress markers (Dogukan et al., 2011) and generates reactive oxygen species (ROS) such as superoxide anion and hydroxyl radical (Kruidering et al., 1997). Increased

Conflict of Interest

There are no conflicts of interest to report for the authors.

Acknowledgments

We thank O.M. Alian (Wayne State University) for carefully editing this manuscript.

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