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. 2019 Aug 1;71(5):905–914. doi: 10.1007/s10616-019-00333-2

Alpha-linolenic acid confers protection on mice renal cells against cisplatin-induced nephrotoxicity

Erman Salih İstifli 1,✉, Erkan Demir 2, Halil Mahir Kaplan 3, Kıvılcım Eren Ateş 4, Figen Doran 4
PMCID: PMC6787136  PMID: 31372877

Abstract

Cisplatin is an antineoplastic agent used in the treatment of various types of solid tumors. Despite the dose-dependency of its antineoplastic effect, the high risk for nephrotoxicity frequently precludes the use of higher doses. α-Linolenic acid (ALA), a carboxylic acid having three cis double bonds, is an essential fatty acid required for health and can be acquired via foods that contain ALA or supplementation of foods high in ALA. Previous studies have shown that ALA demonstrates anti-cancer, anti-inflammatory, and anti-oxidative effects. In this study, we show the protective effect of ALA on cisplatin-induced renal toxicity associated with oxidative stress in mice using biochemical parameters. The mice were randomly assigned into four experimental groups. Group 1 (control group) were administered physiological saline solution for 9 days; group 2 (ALA group) received 200 mg/kg alpha-linolenic acid via gavage for 9 days; group 3 (CIS group) received 100 mg/kg intraperitoneal (i.p.) CIS for 9 days; and group 4 (ALA + CIS group) received 100 mg/kg i.p. CIS and followed by ALA 200 mg/kg via gavage for 9 days. Alpha-linolenic acid significantly reduced the expression of myeloperoxidase (MPO), phospholipase A2 (PLA2), cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in the ALA + CIS group compared to the CIS group. Furthermore, catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) quantities were significantly elevated in the ALA + CIS group when compared to the CIS group. ALA significantly decreased the levels of Bax and cleaved caspase-3, while significantly increasing the level of bcl-2, an anti-apoptotic protein, in the ALA + CIS group than in the CIS group. Finally, histopathological examination in renal tissue showed that the significant edematous damage induced by CIS administration alone was reduced in ALA + CIS group. In conclusion, our findings show that ALA is beneficial to CIS-induced nephrotoxicity in mice via its anti-inflammatory and anti-oxidative effects.

Keywords: Alpha-Linolenic acid, Cisplatin, Oxidative stress, Nephrotoxicity

Introduction

Cisplatin (CIS) is an antineoplastic drug used in the treatment of bladder, small cell and non-small cell lung, head and neck, testis, ovarian, cervix and testicular cancers as well as some other types of cancer (Miller et al. 2010; Oh et al. 2014). CIS acts by cross-linking with the purine bases on DNA, inducing irreversible DNA damage and, consequently, activates apoptosis in cancer cells (Dasari and Tchounwou 2014; Florea and Büsselberg 2011). CIS is a potent chemotherapeutic agent, however, its use has been limited due to serious side effects, particularly organ toxicity. The most significant side effect of CIS is nephrotoxicity (Hartmann et al. 1999). Approximately 20% of patients who receive high-dose CIS treatment develop serious renal failure, and roughly one-third of patients develop renal damage within a few days of initial treatment (Yao et al. 2007).

In many studies, it has been reported that CIS-induced nephrotoxic injury is mediated by oxidative stress, apoptosis and inflammation (Vyas et al. 2014). In kidney cells, CIS initially forms intra and interstrand cross-links with DNA, and binds with the sulfhydryl group of reduced glutathione (GSH) resulting in decreased intracellular levels of free GSH (Mistry et al. 1989). The upregulation of inducible nitric oxide synthase (iNOS) expression by CIS is followed by a reaction between nitric oxide and superoxide anion, leading to the formation of peroxynitrite, which damages cellular biomolecules and mitochondria (Chirino and Pedraza-Chaverri 2009). Moreover, the CIS-induced reduction of the expression of antioxidant enzymes such as glutathione peroxidase (GPx), superoxide dismutase (SOD) and catalase (CAT) contributes to the prolonged persistence of reactive oxygen species (ROS) in the cell (Kadikoylu et al. 2004). Nevertheless, the DNA damage and oxidative stress induced by CIS seems to be independent of each other. CIS-induced DNA damage leads to the translocation of Bax from the cytosol to the mitochondria, an important step in the release of cytochrome c (Cyt c) to the cytosol and the activation of caspases (9 and 3) (Shi 2004).

Among the alternative treatments, omega-3 (n − 3) polyunsaturated fatty acids (PUFA) have been extensively used for their protective roles in oxidative and inflammatory conditions (Calder 2013; Simopoulos 2002). A potential mechanism of action of PUFAs have been proposed previously in which they act as a “sink” to trap free radicals, hence becoming oxidized themselves (Richard et al. 2008).

α-Linolenic acid (ALA) (9Z, 12Z, 15Z)-9,12,15-octadecatrienoic acid) is an n − 3 PUFA essential for health and cannot be produced within the human body. Previous studies have shown that ALA is a protective nutrient against oxidative stress, apoptosis and inflammation (Hassan et al. 2010; Kaplan et al. 2016, 2017). The positive effect of ALA on oxidative stress has been demonstrated by the normalization of intracellular GSH concentrations and decreased iNOS expression. ALA also has a protective effect against inflammation by reducing tumor necrosis factor-alpha (TNF-α) secretion and COX-2 expression by blocking NF-κB activation and inhibiting the phosphorylation of MAP kinases in RAW 264.7 macrophages (Hassan et al. 2010; Ren and Chung 2007). Furthermore, ALA protects cells against apoptosis by decreasing active caspase-3 protein levels (Suphioglu et al. 2010). The decrease in the activity of active caspase-3 has been shown to be related to the protective effect of ALA on cells against apoptosis by inhibiting the production of specific pro-apoptotic oxidized phosphatidylcholine (OxPC) species (Ganguly et al. 2018) as well as increasing the anti-apoptotic protein Bcl-2 levels and thus shifting the Bcl-2/Bax ratio to higher levels (Carotenuto et al. 2016).

Therefore, this study was designed to evaluate the protective effects of ALA on CIS-induced nephrotoxicity in mice for the first time using oxidative stress (MDA, SOD, CAT and GPx), inflammation (myeloperoxidase, phospholipase A2, cyclooxygenase-2 and iNOS) and apoptotic (bcl-2, Bax, cleaved caspase-3) parameters.

Materials and methods

Chemicals

Cisplatin and alpha-linolenic acid (all-cis-9,12,15-octadecatrienoic acid) were purchased from Sigma-Aldrich (St. Louis, Missouri, USA) (Figs. 1, 2).

Fig. 1.

Fig. 1

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Chemical structure of cisplatin

Fig. 2.

Fig. 2

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Chemical structure of alpha-linolenic acid

Assay protocol

In this study, 32 male BALB/c albino mice (30 g) were used. Mice were obtained from Cukurova University Medical Sciences Experimental Research and Application Center (TIBDAM). The Cukurova University Ethics Committee was informed about the protocol to be applied on the experimental animals and approved the protocol for the study prior to performing the experiments. Animals were kept at 20–22 °C and 50–55% humidity for 12 h light/12 h dark cycle. Food and water were supplied ad libitum. The mice were then arbitrarily assigned to four groups. Group 1 (control group, n = 8) was administered physiological saline (0.09% NaCl) solution for 9 days; group 2 (ALA group, n = 8) received 200 mg/kg alpha-linolenic acid by gavage for 9 days; group 3 (CIS group, n = 8), received 100 mg/kg intraperitoneal (i.p.) CIS for 9 days; and group 4 (ALA + CIS, n = 8) received 100 mg/kg i.p. CIS and 200 mg/kg ALA via gavage for 9 days. Thus, in the ALA + CIS group, CIS (100 mg/kg) and ALA (200 mg/kg) were given to mice on a daily basis for 9 days.

24 h after the last injection, the animals were killed by cervical dislocation. Renal tissues in Eppendorf tubes were frozen at − 80 °C and stored for later use in quantitative assays. Bax, Bcl-2, CAT, GPx, activated (cleaved) caspase-3, MDA, MPO, SOD, COX-2, iNOS and PLA2 levels were examined in kidney tissue samples.

Quantitative assays

Tissue homogenization

3 ml of RIPA (Radio-Immunoprecipitation Assay) buffer, 30 μl of PMSF (fenylmetanesulfonylfluoride), 30 μl of sodiumvanadate, 30 μl of protease inhibitor were implemented to the tissues that were frozen at − 20 °C in Eppendorf tubes and homogenates were obtained by breaking down the tissues on ice using an ultrasonic disintegrator. The remaining homogenates were centrifuged at 10,000 rpm for 10 min, the supernatants were stored and the precipitates were (pellets) discarded.

Protein quantification

The protein content of homogenized tissues was determined by Bradford method. Using bovine serum albumin 1, 2, 3, 5, 7, 8 and 10 μg/ml concentrations of standards were prepared. Next, 10 μl of each sample in the experimental groups was taken and completed to 100 μl with distilled water. After adding 1 ml of Bradford solution to the standards and samples followed by mixing with vortex, the absorbances were manually measured at 595 nm wavelength in the spectrophotometer. Protein quantification was performed as “μg/μl” according to the standard curve plotted in the Prism software (La Jolla, CA).

Biochemical analysis

Determination of MDA

The MDA content of the homogenates was detected by spectrophotometrically quantitating the amount of thiobarbituric acid reactive matter (TBARS) (Uchiyama and Mihara 1978). 0.6% thiobarbituric acid (1 ml) and 1% phosphoric acid (3 ml) solutions were added to 0.5 ml of plasma and pipetted into a tube. The mixture was heated in boiling water for 45 min, then cooled and the colored adduct was poured into n-butanol (4 ml). The absorbance was read at 532 nm with a spectrophotometer (UV-1601; Shimadzu, Kyoto, Japan) and lipid peroxidation was determined as the amount of lipid peroxidation’s TBARS. Data were indicated as µmol/mg protein.

Determination of MPO

The activity of tissue-associated MPO (U/mg protein) was determined by the Hillegas method (Hillegass et al. 1990) where the kidney tissue samples were homogenized with a 50 mm potassium phosphate buffer (PB, pH 6.0). Homogenated samples were centrifuged for 10 min at 41,400g. Globs were stopped at 50 mm PB containing 0.5% hexylmethylammonium bromide and processed in three periods of freezing and thawing with sonication between periods. For 10 min, the samples were centrifugated at 41,400g and t. 0.3 ml aliquots were added to a 2.3 ml reaction mixture with 20 mm H2O2 solution and 50 mm PB o-dianisidine. The amount of MPO causing an absorbance alteration was read at 460 nm for 3 min. One unit of enzymatic activity was determined as the amount of MPO.

SOD measurement

The SOD measurement (Mn and Cu–Zn) was performed by the Sun method (Sun et al. 1988) which prevents nitroblue-tetrazole degradation with xanthine–xanthine oxidase as a superoxide generator. One unit SOD was interpreted as the concentration of protein capable of inducing a 50% inhibition of the reduction rate of nitrobluetetrazolium. The results were expressed in units per milligram of protein (U/mg protein).

GPx determination

The method of Paglia and Valentine was used to measure the fixation of GPx activity (EC 1.6.4.2) (Paglia 1989). The enzymatic reaction was triggered with the addition of H2O2 to a tube containing NADPH, glutathione reductase, GSH and sodium azide. The change in absorbance was measured with a spectrophotometer at 340 nm. Data were demonstrated as units per milligram of enzyme (U/mg protein).

Measurement of CAT

The activity of CAT was assessed using the method of Aebi (Aebi 1974) in which the determination of the rate constant (k, s − 1) or the decomposition of H2O2 at 240 nm are the main principles. Results were demonstrated as picogram per milliliter (pg/ml).

Enzyme-linked immunosorbent assay (ELISA)

Expression of Bax, Bcl-2, activated caspase-3, cyclooxygenase-2, phospholipase A2 and iNOS enzymes was investigated by ELISA test according to the manufacturer’s protocol. ELISA kits were obtained from R&D Systems, Inc. (Minneapolis, MN) and used in accordance with the manufacturer’s instructions.

Histopathological assessment

The kidney specimens were obtained by total nephrectomy from a total of 32 male (100%) mice. The control, ALA, CIS and ALA + CIS groups were each consisted of 8 mice, respectively. Isolated kidney tissues for histopathologic examination were fixed in formalin (3.7%) for 24 h and paraffin embedded 4 µm thick tissue sections were stained with hematoxylen and eosin. Samples in each group were evaluated using an Olympus BX53 microscope.

Statistical analysis

Statistical analysis of data was performed with Graph Pad Prism 3.0 (Graph Pad Software, San Diego, USA). The results were expressed as the mean ± standard error (SEM). Comparisons between groups were performed using One-Way Analysis of Variance (ANOVA), followed by a Bonferonni post hoc test. A p value which is < 0.05 was considered statistically significant.

Results

CIS treatment significantly elevated the MPO levels when compared to the control group (p < 0.05), however, the MPO levels in the ALA + CIS group were detected as significantly lower when compared to the CIS group (p < 0.05) (Fig. 3).

Fig. 3.

Fig. 3

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MPO levels in kidney tissues of mice (n = 8). Data groups were compared with an analysis of variance (ANOVA) followed by Bonferroni multiple comparison tests (*p < 0.05 compared to control group, #p < 0.05 compared to CIS group)

CIS treatment significantly elevated the COX-2 levels when compared to the control group (p < 0.05), however, the COX-2 levels in the ALA + CIS group were detected as significantly lower when compared to the CIS group (p < 0.05) (Fig. 4).

Fig. 4.

Fig. 4

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COX-2 levels in kidney tissues of mice (n = 8). Data groups were compared with an analysis of variance (ANOVA) followed by Bonferroni multiple comparison tests (*p < 0.05 compared to control group, #p < 0.05 compared to CIS group)

The increase in the iNOS levels was found to be statistically significant in the CIS group when compared with the control group (p < 0.05), however, the iNOS levels in the ALA + CIS group were detected as significantly lower when compared to the CIS group (p < 0.05) (Fig. 5).

Fig. 5.

Fig. 5

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iNOS levels in kidney tissues of mice (n = 8). Data groups were compared with an analysis of variance (ANOVA) followed by Bonferroni multiple comparison tests (*p < 0.05 compared to control group, #p < 0.05 compared to CIS group)

PLA2 levels were significantly elevated in the CIS group when compared with the control group (p < 0.05), however, the PLA2 levels were significantly lower in the ALA + CIS group compared to the CIS group (p < 0.05) (Fig. 6).

Fig. 6.

Fig. 6

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PLA2 levels in kidney tissues of mice (n = 8). Data groups were compared with an analysis of variance (ANOVA) followed by Bonferroni multiple comparison tests (*p < 0.05 compared to control group, #p < 0.05 compared to CIS group)

CIS treatment significantly elevated the MDA levels when compared to the control group (p < 0.05), however, the MDA levels in the ALA + CIS group were detected as significantly lower when compared to the CIS group (p < 0.05) (Fig. 7).

Fig. 7.

Fig. 7

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MDA levels in kidney tissues of mice (n = 8). Data groups were compared with an analysis of variance (ANOVA) followed by Bonferroni multiple comparison tests (*p < 0.05 compared to control group, #p < 0.05 compared to CIS group)

CIS treatment significantly decreased the SOD, CAT and GPx levels when compared to the control group (p < 0.05). On the other hand, the levels of these three enzymes were found to be significantly elevated in the ALA + CIS group than those in the CIS group (p < 0.05) (Figs. 8, 9, 10).

Fig. 8.

Fig. 8

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SOD levels in kidney tissues of mice (n = 8). Groups of data were compared with an analysis of variance (ANOVA) followed by Bonferroni multiple comparison tests (*p < 0.05; compared to control group, #p < 0.05; compared to CIS group)

Fig. 9.

Fig. 9

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CAT levels in kidney tissues of mice (n = 8). Groups of data were compared with an analysis of variance (ANOVA) followed by Bonferroni multiple comparison tests (*p < 0.05; compared to control group, #p < 0.05; compared to CIS group)

Fig. 10.

Fig. 10

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GPx levels in kidney tissues of mice (n = 8). Groups of data were compared with an analysis of variance (ANOVA) followed by Bonferroni multiple comparison tests (*p < 0.05; compared to control group, #p < 0.05; compared to CIS group)

CIS treatment significantly increased the Bax and cleaved caspase-3 levels as compared to the control group (p < 0.05), however, co-treatment with ALA (ALA + CIS group) resulted in a significant reduction in the levels of these proteins compared to the CIS group (p < 0.05) (Figs. 11, 12).

Fig. 11.

Fig. 11

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Bax levels in kidney tissues of mice (n = 8). Groups of data were compared with an analysis of variance (ANOVA) followed by Bonferroni multiple comparison tests (*p < 0.05; compared to control group, #p < 0.05; compared to CIS group)

Fig. 12.

Fig. 12

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Cleaved caspase-3 levels in kidney tissues of mice (n = 8). Groups of data were compared with an analysis of variance (ANOVA) followed by Bonferroni multiple comparison tests (*p < 0.05; compared to control group, #p < 0.05; compared to CIS group)

CIS treatment significantly decreased the Bcl-2 levels when compared with the control group (p < 0.05). In contrast, co-treatment with ALA (ALA + CIS group) significantly increased the Bcl-2 levels compared to the CIS group (p < 0.05) (Fig. 13).

Fig. 13.

Fig. 13

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The Bcl-2 levels in kidney tissues of mice (n = 8). Groups of data were compared with an analysis of variance (ANOVA) followed by Bonferroni multiple comparison tests (*p < 0.05; compared to control group, #p < 0.05; compared to CIS group)

The kidney biopsy sample was composed of medullary and cortical areas (Figs. 14, 15). The injury caused by CIS affected mainly interstitial space without glomerular/or tubular damage. The glomerular tufts were normal, however, there was significant interstitial edematous fluid collection (Figs. 16, 17).

Fig. 14.

Fig. 14

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Light micrograph sections of the kidney of control mice. Arrow: glomerulus (renal corpuscle), asterisk: tubulus. Scale bar = 100 µm

Fig. 15.

Fig. 15

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Light micrograph sections of mice treated with ALA. ALA indicated no detrimental effect on kidneys. Arrow: glomerulus (renal corpuscle), asterisk: tubulus. Scale bar = 100 µm

Fig. 16.

Fig. 16

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Light micrograph sections of mice treated with CIS. CIS administration resulted in significant edematous damage (diamonds). Arrow: glomerulus (renal corpuscle), asterisk: tubulus, diamond: edema. Scale bar = 100 µm

Fig. 17.

Fig. 17

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Light micrograph sections of mice treated with ALA + CIS. ALA co-administration ameliorated the kidney damage caused by CIS. Arrow: glomerulus (renal corpuscle), asterisk: tubulus. Scale bar = 100 µm

Discussion

The results of our study showed for the first time that renal damage induced by CIS increased oxidative stress in kidney tissue and that ALA application had a protective effect on these adverse effects induced by CIS.

We evaluated various enzymes acting as biological mediators. MPO activity is a widely used parameter in the characterization and observation of inflammation and the decreased activity of this pro-inflammatory protein has been determined as an indicator of the anti-inflammatory effect of the investigated substance in question (Camuesco et al. 2005). In this study, MPO levels were found to be increased in the CIS treated group. Increased MPO activity shows that neutrophil and monocyte accumulation in kidney tissue contribute to oxidative stress induced by CIS. In contrast, co-treatment with ALA was found to significantly reduce MPO activity in our study.

It has been reported that NO produced by iNOS, another mediator of inflammation we investigated in this study, had a pathogenic role in acute and chronic inflammatory diseases (Camuesco et al. 2005; Poljakovic et al. 2001; Esposito and Cuzzocrea 2007). In addition, COX-2 and the prostaglandins synthesized by this enzyme also play a role in inflammation (Galecki et al. 2012). In our study, co-treatment with ALA caused a significant decrease in the amount of iNOS and COX-2 enzymes increased by CIS alone. The results obtained in our study are consistent with previous studies. ALA isolated from Actinidia polygama significantly inhibited iNOS and COX-2 gene expression and NO production, by directly blocking activation of NF-kB (Ren and Chung 2007).

Studies have shown that increased intracellular calcium levels following the induction of oxidative stress leads to an increase in the activity of PLA2 (Malis and Bonventre 1986; Rordorf et al. 1991). In our study, it was determined that ALA decreased the amount of PLA2 enzyme increased by CIS treatment alone. ALA has been shown to have no direct effect on the intracellular activity or the mass of the PLA2 enzyme (Nelson et al. 2011). The ALA molecule we used in our study is energetically compatible to oxidation due to the double bonds it possesses in its structure and therefore has potential antioxidative properties. Thus, ALA may have indirectly inhibited the increase of the activity of PLA2 enzyme in our study. In accordance with this hypothesis, Pedersen et al. showed that the decrease in the amount of PLA2 was not due to the direct effect of ALA on the enzyme, but rather it reduced the oxidation of the substrate (LDL-C) of PLA2 (Nelson et al. 2011).

Oxidative stress plays an important role in the nephrotoxic damage induced by CIS. CIS causes decrease in the GSH levels by forming adducts with glutathione and increases lipid peroxidation (Chirino and Pedraza-Chaverri 2009). MDA is an oxidative stress marker and is the product of lipid peroxidation induced by free radicals. In our study, CIS caused a significant increase in MDA level in kidney tissue. The results of our study are consistent with previous studies reporting the toxic effect of CIS by increasing MDA levels in renal cells (Zhou et al. 2006; Verma et al. 2016). In our study, the MDA level was significantly decreased in the ALA + CIS group compared to the CIS-treated group.

In this study, GPx, SOD and CAT levels were significantly decreased in the CIS treated group compared to the control group, however, co-treatment with ALA (ALA + CIS group) was found to significantly increase the levels of these three enzymes that play a protective role from oxidative stress. Our findings are consistent with a study reporting that CIS significantly reduces GPx, SOD and CAT activity in kidney tissue (Maliakel et al. 2008). There are no mechanism-based studies underlying the ameliorative effect of ALA on GPx, SOD and CAT activity, but the protective effect provided by ALA against the toxicity of CIS can be attributed to the intrinsic biochemical and natural antioxidant properties of this substance.

In our study, CIS increased the levels of pro-apoptotic proteins bax and caspase-3 and decreased the amount of anti-apoptotic protein bcl-2. However, ALA application reversed these effects. The Bcl-2 family and caspases are the main mediators in the apoptotic pathway. While the intrinsic apoptotic pathway induces apoptosis by directly activating caspase-3 via the formation of apoptosome complex, the extrinsic pathway activates caspase-3 through the death inducing signaling complex (DISC) formation and activation of caspases-8 and -10. In this last step, cleaved caspase-3 causes typical DNA fragmentation. The Bcl-2 family has both apoptotic and anti-apoptotic members. Proapotic or anti-apoptotic signals, which are formed according to the balance between Bcl-2 family proteins, affect mitochondria. When the apoptotic signals are dominant, the cytochrome c is released from the mitochondria to form the apoptosome complex. Triggered apoptosis causes organ failure, whereas apoptosis inhibition causes hyperplasia and cancer. Apoptosis defects are important in developmental, autoimmune and neurodegenerative diseases and cancer development (Kaplan et al. 2017).

Oxidative stress in nephrons plays role in the induction of apoptosis and necrosis by increasing the levels of inflammatory mediators (TNF-α, IL-6, NF-kB and MPO) and apoptotic proteins (Bax, caspase-3 and caspase-9) (Sahu et al. 2014). Our findings indicate that CIS causes apoptosis and necrosis in nephrons and suggest that ALA can prevent nephrotoxicity induced by CIS in nephrons by inhibiting both apoptosis and necrosis pathways.

In conclusion, our study showed that ALA, an n-3 fatty acid, has a protective effect in mice renal tissue. The protective effect of ALA against nephrotoxicity induced by CIS may be related to its anti-oxidant, anti-apoptotic and anti-inflammatory properties. Our results indicate that ALA is a promising molecule in nephrotoxicity induced by CIS.

Funding acknowledgement

This study has been funded by Cukurova University Scientific Research Projects Unit (FBA-2018-10148).

Footnotes

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

References

  1. Aebi H. Catalase. In: Bergmeyer HU, editor. Methods of enzymatic analysis. New York: Academic Press; 1974. pp. 673–677. [Google Scholar]
  2. Calder PC. Omega-3 polyunsaturated fatty acids and inflammatory processes: nutrition or pharmacology? Br J Clin Pharmacol. 2013;75:645–662. doi: 10.1111/j.1365-2125.2012.04374.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Camuesco D, Galvez J, Nieto A, Comalada M, Rodriguez-Cabezas ME, Concha A, Xaus J, Zarzuelo A. Dietary olive oil supplemented with fish oil, rich in EPA and DHA (n-3) polyunsaturated fatty acids, attenuates colonic inflammation in rats with DSS-induced colitis. J Nutr. 2005;135:687–694. doi: 10.1093/jn/135.4.687. [DOI] [PubMed] [Google Scholar]
  4. Carotenuto F, Coletti D, Di Nardo P, Teodori L. α-Linolenic acid reduces TNF-induced apoptosis in C2C12 myoblasts by regulating expression of apoptotic proteins. Eur J Transl Myol. 2016;26:6033. doi: 10.4081/ejtm.2016.6033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chirino YI, Pedraza-Chaverri J. Role of oxidative and nitrosative stress in cisplatin-induced nephrotoxicity. Exp Toxicol Pathol. 2009;61:223–242. doi: 10.1016/j.etp.2008.09.003. [DOI] [PubMed] [Google Scholar]
  6. Dasari S, Tchounwou PB. Cisplatin in cancer therapy: molecular mechanisms of action. Eur J Pharmacol. 2014;740:364–378. doi: 10.1016/j.ejphar.2014.07.025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Esposito E, Cuzzocrea S. The role of nitric oxide synthases in lung inflammation. Curr Opin Invest Dr. 2007;8:899–909. [PubMed] [Google Scholar]
  8. Florea AM, Büsselberg D. Cisplatin as an anti-tumor drug: cellular mechanisms of activity, drug resistance and induced side effects. Cancers. 2011;3:1351–1371. doi: 10.3390/cancers3011351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Galecki P, Galecka E, Maes M, Chamielec M, Orzechowska A, Bobinska K, Lewinski A, Szemraj J. The expression of genes encoding for COX-2, MPO, iNOS, and sPLA2-IIA in patients with recurrent depressive disorder. J Affect Disord. 2012;138:360–366. doi: 10.1016/j.jad.2012.01.016. [DOI] [PubMed] [Google Scholar]
  10. Ganguly R, Hasanally D, Stamenkovic A, Maddaford TG, Chaudhary R, Pierce GN, Ravandi A. Alpha linolenic acid decreases apoptosis and oxidized phospholipids in cardiomyocytes during ischemia/reperfusion. Mol Cell Biochem. 2018;437:163–175. doi: 10.1007/s11010-017-3104-z. [DOI] [PubMed] [Google Scholar]
  11. Hartmann JT, Kollmannsberger C, Kanz L, Bokemeyer C. Platinum organ toxicity and possible prevention in patients with testicular cancer. Int J Cancer. 1999;83:866–869. doi: 10.1002/(SICI)1097-0215(19991210)83:6&#x0003c;866::AID-IJC34&#x0003e;3.0.CO;2-9. [DOI] [PubMed] [Google Scholar]
  12. Hassan A, Ibrahim A, Mbodji K, Coeffier M, Ziegler F, Bounoure F, Chardigny JM, Skiba M, Savoye G, Dechelotte P, Marion-Letellier R. An alpha-linolenic acid-rich formula reduces oxidative stress and inflammation by regulating NF-kappaB in rats with TNBS-induced colitis. J Nutr. 2010;140:1714–1721. doi: 10.3945/jn.109.119768. [DOI] [PubMed] [Google Scholar]
  13. Hillegass LM, Griswold DE, Brickson B, Albrightsonwinslow C. Assessment of myeloperoxidase activity in whole rat-kidney. J Pharmacol Method. 1990;24:285–295. doi: 10.1016/0160-5402(90)90013-B. [DOI] [PubMed] [Google Scholar]
  14. Kadikoylu G, Bolaman Z, Demir S, Balkaya M, Akalin N, Enli Y. The effects of desferrioxamine on cisplatin-induced lipid peroxidation and the activities of antioxidant enzymes in rat kidneys. Hum Exp Toxicol. 2004;23:29–34. doi: 10.1191/0960327104ht413oa. [DOI] [PubMed] [Google Scholar]
  15. Kaplan HM, Izol V, Arıdogan IA, Olgan E, Yegani AA, Pazarcı P, Singirik E. Protective effect of alpha-linolenic acid on gentamicin induced nephrotoxicity in mice. Int J Pharm. 2016;12:562–566. doi: 10.3923/ijp.2016.562.566. [DOI] [Google Scholar]
  16. Kaplan HM, Singirik E, Erdogan KE, Doran F. Protective effect of alpha-linolenic acid on gentamicin-induced ototoxicity in mice. Somatosens Mot Res. 2017;34:145–150. doi: 10.1080/08990220.2017.1356283. [DOI] [PubMed] [Google Scholar]
  17. Maliakel DM, Kagiya TV, Nair CK. Prevention of cisplatin-induced nephrotoxicity by glucosides of ascorbic acid and alpha-tocopherol. Exp Toxicol Pathol. 2008;60:521–527. doi: 10.1016/j.etp.2008.04.015. [DOI] [PubMed] [Google Scholar]
  18. Malis CD, Bonventre JV. Mechanism of calcium potentiation of oxygen free radical injury to renal mitochondria. A model for post-ischemic and toxic mitochondrial damage. J Biol Chem. 1986;261:14201–14208. [PubMed] [Google Scholar]
  19. Miller RP, Tadagavadi RK, Ramesh G, Reeves WB. Mechanisms of cisplatin nephrotoxicity. Toxins (Basel) 2010;2:2490–2518. doi: 10.3390/toxins2112490. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mistry P, Lee C, McBrien DC. Intracellular metabolites of cisplatin in the rat kidney. Cancer Chemother Pharmacol. 1989;24:73–79. doi: 10.1007/BF00263124. [DOI] [PubMed] [Google Scholar]
  21. Nelson TL, Hokanson JE, Hickey MS. Omega-3 fatty acids and lipoprotein associated phospholipase A2 in healthy older adult males and females. Eur J Nutr. 2011;50:185–193. doi: 10.1007/s00394-010-0126-7. [DOI] [PubMed] [Google Scholar]
  22. Oh GS, Kim HJ, Shen A, Lee SB, Khadka D, Pandit A, So HS. Cisplatin-induced kidney dysfunction and perspectives on improving treatment strategies. Electrolyte Blood Press. 2014;12:55–65. doi: 10.5049/EBP.2014.12.2.55. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Paglia DE. Erythroenzymopathies—1960s in retrospect—a citation classic commentary on studies on the quantitative and qualitative characterization of erythrocyte glutathione-peroxidase by Paglia DE. Clin Med. 1989;8:16. [PubMed] [Google Scholar]
  24. Poljakovic M, Svensson ML, Svanborg C, Johansson K, Larsson B, Persson K. Escherichia coli-induced inducible nitric oxide synthase and cyclooxygenase expression in the mouse bladder and kidney. Kidney Int. 2001;59:893–904. doi: 10.1046/j.1523-1755.2001.059003893.x. [DOI] [PubMed] [Google Scholar]
  25. Ren J, Chung SH. Anti-inflammatory effect of alpha-linolenic acid and its mode of action through the inhibition of nitric oxide production and inducible nitric oxide synthase gene expression via NF-kappaB and mitogen-activated protein kinase pathways. J Agric Food Chem. 2007;55:5073–5080. doi: 10.1021/jf0702693. [DOI] [PubMed] [Google Scholar]
  26. Richard D, Kefi K, Barbe U, Bausero P, Visioli F. Polyunsaturated fatty acids as antioxidants. Pharmacol Res. 2008;57:451–455. doi: 10.1016/j.phrs.2008.05.002. [DOI] [PubMed] [Google Scholar]
  27. Rordorf G, Koroshetz WJ, Bonventre JV. Heat shock protects cultured neurons from glutamate toxicity. Neuron. 1991;7:1043–1051. doi: 10.1016/0896-6273(91)90348-4. [DOI] [PubMed] [Google Scholar]
  28. Sahu BD, Tatireddy S, Koneru M, Borkar RM, Kumar JM, Kuncha M, Srinivas R, Sunder RS, Sistla R. Naringin ameliorates gentamicin-induced nephrotoxicity and associated mitochondrial dysfunction, apoptosis and inflammation in rats: possible mechanism of nephroprotection. Toxicol Appl Pharm. 2014;277:8–20. doi: 10.1016/j.taap.2014.02.022. [DOI] [PubMed] [Google Scholar]
  29. Shi Y. Caspase activation, inhibition, and reactivation: a mechanistic view. Protein Sci. 2004;13:1979–1987. doi: 10.1110/ps.04789804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Simopoulos AP. Omega-3 fatty acids in inflammation and autoimmune diseases. J Am Coll Nutr. 2002;21:495–505. doi: 10.1080/07315724.2002.10719248. [DOI] [PubMed] [Google Scholar]
  31. Sun Y, Oberley LW, Li Y. A simple method for clinical assay of superoxide dismutase. Clin Chem. 1988;34:497–500. [PubMed] [Google Scholar]
  32. Suphioglu C, De Mel D, Kumar L, Sadli N, Freestone D, Michalczyk A, Sinclair A, Ackland ML. The omega-3 fatty acid, DHA, decreases neuronal cell death in association with altered zinc transport. FEBS Lett. 2010;584:612–618. doi: 10.1016/j.febslet.2009.12.013. [DOI] [PubMed] [Google Scholar]
  33. Uchiyama M, Mihara M. Determination of malonaldehyde precursor in tissues by thiobarbituric acid test. Anal Biochem. 1978;86:271–278. doi: 10.1016/0003-2697(78)90342-1. [DOI] [PubMed] [Google Scholar]
  34. Verma PK, Raina R, Sultana M, Singh M, Kumar P. Total antioxidant and oxidant status of plasma and renal tissue of cisplatin-induced nephrotoxic rats: protection by floral extracts of Calendula officinalis Linn. Ren Fail. 2016;38:142–150. doi: 10.3109/0886022X.2015.1103585. [DOI] [PubMed] [Google Scholar]
  35. Vyas D, Laput G, Vyas AK. Chemotherapy-enhanced inflammation may lead to the failure of therapy and metastasis. Oncotargets Ther. 2014;7:1015–1023. doi: 10.2147/OTT.S60114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Yao X, Panichpisal K, Kurtzman N, Nugent K. Cisplatin nephrotoxicity: a review. Am J Med Sci. 2007;334:115–124. doi: 10.1097/MAJ.0b013e31812dfe1e. [DOI] [PubMed] [Google Scholar]
  37. Zhou H, Kato A, Miyaji T, Yasuda H, Fujigaki Y, Yamamoto T, Yonemura K, Takebayashi S, Mineta H, Hishida A. Urinary marker for oxidative stress in kidneys in cisplatin-induced acute renal failure in rats. Nephrol Dial Transplant. 2006;21:616–623. doi: 10.1093/ndt/gfi314. [DOI] [PubMed] [Google Scholar]

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