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Attenuation of Gentamycin-induced nephrotoxicity in rats by dietary inclusion of ginger ( Zingiber officinale) and turmeric (Curcuma longa) rhizomes Adedayo O Ademiluyi, Ganiyu Oboh, Opeyemi B Ogunsuyi and Ayodele J. Akinyemi Nutrition and Health published online 5 November 2013 DOI: 10.1177/0260106013506668 The online version of this article can be found at: http://nah.sagepub.com/content/early/2013/10/17/0260106013506668 A more recent version of this article was published on - Mar 11, 2014
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Article
Attenuation of gentamycin-induced nephrotoxicity in rats by dietary inclusion of ginger (Zingiber officinale) and turmeric (Curcuma longa) rhizomes
Nutrition and Health 1-10 ª The Author(s) 2013 Reprints and permission: sagepub.co.uk/journalsPermissions.nav DOI: 10.1177/0260106013506668 nah.sagepub.com
Adedayo O Ademiluyi1, Ganiyu Oboh1, Opeyemi B Ogunsuyi1, and Ayodele J Akinyemi1,2
Abstract This study sought to investigate the modulatory effects of dietary inclusion of ginger (Zingiber officinale) and turmeric (Curcuma longa) rhizomes on antioxidant status and renal damage induced by gentamycin in rats. Renal damage was induced in albino rats pretreated with dietary inclusion of ginger and turmeric (2% and 4%) by intraperitoneal (i.p.) administration of gentamycin (100 mg/kg body weight) for three days. Assays for renal damage biomarkers (plasma creatinine, plasma urea, blood urea nitrogen and plasma uric acid), malondialdehyde (MDA) content and reduced glutathione (GSH) content as well as renal antioxidant enzymes (catalase, glutathione-Stransferase (GST), glutathione peroxidase (GPx) and superoxide dismutase (SOD)) were carried out. The study revealed significant (p < 0.05) increases in renal damage biomarkers following gentamycin administration with severe alteration in kidney antioxidant status. However, pretreatment with ginger and turmeric rhizome (2% and 4%) prior to gentamycin administration significantly (p < 0.05) protected the kidney and attenuated oxidative stress by modulating renal damage and antioxidant
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Functional Foods and Nutraceuticals Unit, Federal University of Technology, Akure, Nigeria Department of Biochemistry, Afe Babalola University, Ado-Ekiti, Nigeria
Corresponding author: Adedayo O Ademiluyi, Functional Foods and Nutraceuticals Unit, Department of Biochemistry, Federal University of Technology, Akure P.M.B. 704, Akure, 340001, Nigeria. Email: [email protected]
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indices. This finding therefore suggests that dietary inclusion of ginger and turmeric rhizomes may protect against gentamycin-induced nephrotoxicity and oxidative stress. Keywords Ginger, Zingiber officinale, turmeric, Curcuma longa, gentamycin, nephrotoxicity, oxidative stress
Introduction The kidneys are vital organs essential for excretion of metabolic wastes as well as maintaining chemical homeostasis among other functions. Many research have linked oxidative stress as a potential cause of different forms of renal damage or nephrotoxicity. Studies have shown that increase in oxidative stress due to free radical generation is a likely result of inflammatory responses associated with patients suffering from acute renal failure (Himmelfarb et al., 2004), while accumulated evidences give credence to the fact that there is elevated level of oxidative stress in patients with chronic renal failure which is often accompanied by proportional endothelial cell damage (Annuk et al., 2001). Considering the vital physiological functions of the kidney and the unique role played by free radical-induced oxidative stress in different forms of acute and chronic renal damage, it is important to explore ways by which oxidative stress may be ameliorated. Xenobiotic interaction with renal tubular cells is one way of generating free radicals in the kidney. Gentamycin is a bacteriocidal antibiotic (Martı´nez-Salgado et al., 2007) used mainly against gram-negative bacteria (Moulds and Jeyasingham, 2010). However, this drug has some side effects with nephrotoxicity and ototoxicity being most studied (Martı´nez-Salgado et al., 2007). Nephrotoxicity resulting from gentamycin has been established at a dose above 2 mg/ml in humans (Shaibu and Chika, 2009). Studies have suspected that gentamycin induces free radical generation in renal tubular cells to elicit nephrotoxicity (Martı´nez-Salgado et al., 2007). Gentamycin is concentrated in the renal proximal tubular cells where it induces generation of superoxide anions, hydrogen peroxide and hydroxyl radicals. However, a lack of detailed understanding of the mechanism of toxicity of gentamycin has made it difficult to control its toxicity (Shaibu and Chika, 2009). Nevertheless, the use of some synthetic antioxidant compounds/molecules to counteract the effects of free radical generation has been suggested as a viable management strategy, though their attendant side effects are a major drawback, hence the need for alternatives having little or no side effects, with plant sources being the most promising. Spices have been widely used as food adjunct and in folk medicine for several centuries (Adefegha and Oboh, 2012). Ginger (Zingiber officinale) rhizome is used widely as a spice for flavoring foods and has wide applications in folk medicine. Studies have shown its anti-inflammatory, hypoglycemic and hypolipidemic properties (Abdullahi, 2011; Ahmad, 1997). The antioxidant property of Z. officinale has been linked to its 2 Downloaded from nah.sagepub.com at GEORGIAN COURT UNIV on October 20, 2014
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volatile oil, polyphenol and flavonoid content (Ali et al., 2010; Lakshmi and Sudhakar, 2010). Furthermore, turmeric (Curcuma longa) which is another rhizomatous plant belonging to the ginger family has been employed as a dye source and food colorant due to its color, and it is also used as a food flavoring agent. Curcuminoids are the major phytochemicals of turmeric which are responsible for the characteristic yellow color (Jurenka, 2009). Curcumin, one of the predominant curcuminoids and a flavonoid, has been investigated for anti-inflammatory and antioxidant properties (Hatcher et al., 2008). Curcumin has also been studied to ‘mop up’ superoxide anions, peroxynitrite radicals and singlet oxygen (Biswas et al., 2005). The ability to derive cheap dietary sources of antioxidants in ginger and turmeric offers a potential dual benefit for their use as culinary spices and to ameliorate oxidative stress. Thus, antioxidant sources with no known toxicological side effects (at safe dosage) offer a good way to manage nephrotoxicity generated by the use of gentamycin.
Materials and method Materials Fresh rhizomes of ginger and turmeric were purchased from Oja Oba market in Akure, Ondo State Nigeria. The authentication of the samples was done in the Department of Crop, Soil and Pest Management, Federal University of Technology, Akure, Nigeria. These samples were washed, chopped and sun-dried to dryness after which they were ground into fine powder. All chemicals and reagents used were of analytical grade and distilled water was used. All the kits used for bioassays were sourced from Randox Laboratories Ltd., Crumlin, Co. Antrim, UK. Gentamycin was sourced as Enagent Gentamycin Sulphate Injection (Zhejang, JinlingTiafeng Pharmaceutical Factory, Chezhan Road, Huzhou City, Zhejiiang, China). Diet ingredients were purchased from VITAL Feeds, Jos, Nigeria Ltd.
Animals Male albino rats weighing 97–121 g used for this experiment were purchased from the breeding colony of a private farm in Akure, Ondo State, Nigeria. The rats were maintained at 25oC on a 12 h light/dark cycle with unrestricted access to food and water. They were acclimatized under these conditions for two weeks prior to the commencement of the experiments. This study received the approval of the institution’s Ethical Committee on the use of laboratory animals and the animals were handled in accordance with National Institute of Health (NIH) guide for the care and use of laboratory animals.
Experimental design and induction of nephrotoxicity After acclimatization, the rats were randomly divided into six groups of six animals each. Group I and Group II were fed a basal diet (50% skimmed milk, 36% corn starch, 10% groundnut oil and 4% mineral & vitamin premix) following a slightly modified method of Oboh et al. (2010), Groups III and IV were fed basal diet supplemented with 2% and 4% ginger rhizome respectively and groups V and VI were fed basal diet supplemented 3 Downloaded from nah.sagepub.com at GEORGIAN COURT UNIV on October 20, 2014
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with 2% and 4% turmeric respectively for 27 days prior to gentamycin (100 mg/kg body weight i.p.) administration (Bushra and Effat, 2007) which lasted for three days and the experiment was terminated after 30 days. The rats were subject to an overnight fast after which they were decapitated by cervical dislocation. The blood was rapidly collected into separate Ethylene diaminetetraacetic acid (EDTA) bottles by direct heart puncture and centrifuged at 800g for 10 min to separate the plasma. Similarly, the kidneys were isolated, rinsed in cold saline (0.9% NaCl) and homogenized in phosphate buffer (pH 6.9). The homogenates were centrifuged at 5000g for 10 min to obtain the clear supernatant. The clear supernatant obtained was used for various biochemical assays (Belle et al., 2004).
Analytical procedures The assay for the renal damage biomarkers; plasma creatinine, plasma urea and plasma uric acid were carried out according to the manufacturer’s procedure of the commercially available kits (Randox Laboratories UK). Blood urea nitrogen (BUN) was subsequently calculated from the plasma urea concentration. Lipid peroxidation was determined as reported by Ohkawa et al. (1979) with slight modification; catalase (CAT) activity was assayed as reported by Sinha (1972); glutathione-S-transferase (GST) activity was assayed according to Habig et al., (1974); glutathione peroxidase (GPx) activity was determined by the method of Rotruck et al. (1973); superoxide dismutase (SOD) activity was determined by the method of Alia et al. (2003); reduced glutathione (GSH) content was determined by the modified method of Ellman (1959) and total protein was determined according to Lowry et al. (1951).
Statistical analysis The results of replicate experiments were represented as mean+standard deviation (SD). Statistical analyses were done using statistical program for social science (SPSS) 16.0 (SPSS. Inc. Chicago, Illinois, USA). One way analysis of variance (ANOVA) was carried out followed by Duncan’s test for the post-hoc treatment. A value of p < 0.05 was considered to be statistically significant.
Results The percentage average weight gain/loss of the rats in each treatment groups is presented in Table 1. Weights were recorded before feeding the rats with supplemented diets (Day 1) and 27 days after feeding them with the supplemented diets. The result revealed a significant (p < 0.05) percentage weight increase across the six groups of experimental rats after feeding on the formulated diets for 27 days. However, three days after gentamycin (100 mg/kg body weight i.p.) administration (Day 30) to the rats, a significant weight decrease was recorded in the control rats. Nevertheless, the groups fed diets supplemented with ginger and turmeric rhizome (2% and 4%) recorded significant weight gain compared to the control rats (except those with 2% turmeric inclusion). 4 Downloaded from nah.sagepub.com at GEORGIAN COURT UNIV on October 20, 2014
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Table 1. Effect of dietary inclusion of ginger and turmeric rhizome on average weight gain/loss in gentamycin-administered rats. Weight (g/rat) Groups I II III IV V VI
Day 1
Day 27
165.8+39.6 150.8+52.2 84.9+70.0 91.3+56.4 143.4+60.9 107.4+37.5
170.6+29.9 167.3+31.0 90.3+31.0 95.4+46.1 158.7+35.2 138.7+2.6
Weight (g/rat) Weight gain/loss (%) a
2.9 10.9c 6.3b 4.5b 10.7c 29.1d
Day 30
Weight gain/loss (%)
192.9+40.0 148.9+29.6 95.3+51.8 99.7+44.2 147.2+37.3 157.6+18.9
13.1c –11.0a 5.5b 4.5b –7.6ab 13.7c
I: normal rats fed with basal diet; II: control rats (gentamycin-administered rats fed with basal diet); III: gentamycin-administered rats fed with diet supplemented with 2% ginger rhizome; IV: gentamycinadministered rats fed with diet supplemented with 4% ginger rhizome; V: gentamycin-administered rats fed with diet supplemented with 2% turmeric; VI: gentamycin-administered rats fed with diet supplemented with 4% turmeric rhizome. Values represent mean+standard deviation (n ¼ 6). Values with the same superscript number on the same row are not significantly (p < 0.05) different.
Table 2. Effect of dietary inclusion of ginger and turmeric rhizome on renal damage biomarkers in gentamycin-administered rats. Plasma concentration of renal damage biomarkers (mg/dl) Groups I II III IV V VI
Creatinine
Urea
BUN
Uric acid
0.25+0.05a 0.36+0.13c 0.34+0.01bc 0.32+0.04bc 0.31+0.08abc 0.29+0.05ab
67.7+6.4a 77.2+2.0c 72.1+6.9b 66.9+6.2a 71.1+2.5b 73.1+2.7c
31.6+3.0a 36.1+0.9c 33.7+3.2b 31.3+2.9a 33.2+1.2b 34.1+1.3c
5.4+1.0bc 7.2+0.8d 6.0+0.5cd 2.8+1.3a 3.9+0.1b 5.8+0.8cd
I: normal rats fed with basal diet; II: control rats (gentamycin-administered rats fed with basal diet); III: gentamycin-administered rats fed with diet supplemented with 2% ginger rhizome; IV: gentamycinadministered rats fed with diet supplemented with 4% ginger rhizome; V: gentamycin-administered rats fed with diet supplemented with 2% turmeric; VI: gentamycin-administered rats fed with diet supplemented with 4% turmeric rhizome. Values represent mean+standard deviation (n ¼ 6). Values with the same superscript number on the same row are not significantly (p < 0.05) different
Table 2 shows the effect of the dietary inclusion of ginger and turmeric rhizome on the renal damage biomarkers (plasma creatinine, urea, BUN and uric acid) in normal rats and gentamycin-administered rats. The result shows that the plasma level of creatinine significantly (p < 0.05) increases in gentamycin-administered (control) rats (0.36 mg/ dl) when compared with the normal rats (0.25 mg/dl). However, the plasma creatinine level was significantly (p < 0.05) restored in gentamycin-administered rats fed diets supplemented with ginger and turmeric rhizome (2% and 4%). Similarly, the plasma urea 5 Downloaded from nah.sagepub.com at GEORGIAN COURT UNIV on October 20, 2014
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Table 3. Effect of dietary inclusion of ginger and turmeric rhizome on tissue enzymatic and nonenzymatic antioxidant indices in gentamycin-administered rats. Groups I II III IV V VI
Catalase U/g protein
GST U/g protein
5.4+0.4bc 4.4+0.2a 5.9+0.5c 7.0+0.3d 5.0+0.7b 5.0+0.3ab
3.0+0.6c 2.0+0.4ab 1.7+0.2a 2.9+0.6bc 2.6+0.5bc 2.8+0.5bc
GPx U/g protein
SOD U/g protein
1.1+0.032a 11.3+3.6c 0.9+ 0.23c 6.9+1.8abc 0.9+ 0.27b 6.2+1.0ab 1.2+ 0.31a 4.7+1.8a 1.3+ 0.18b 9.2+3.0c 1.7+ 0.23c 8.2+1.7bc
MDA mmol/g protein
GSH mg/g protein
2.4+0.9a 3.5+0.7b 2.5+0.1a 2.4+0.5a 2.3+0.9a 2.2+0.1a
13.8+2.3a 10.1+1.6a 12.8+2.0a 13.1+3.1a 12.9+1.5a 13.0+2.0a
GPx: glutathione peroxidase; GSH: reduced glutathione; GST: glutathione-S-transferase; MDA: malondialdehyde; SOD: superoxide dismutase. I: normal rats fed with basal diet; II: control rats (gentamycin-administered rats fed with basal diet); III: gentamycin-administered rats fed with diet supplemented with 2% ginger rhizome; IV: gentamycinadministered rats fed with diet supplemented with 4% ginger rhizome; V: gentamycin-administered rats fed with diet supplemented with 2% turmeric; VI: gentamycin-administered rats fed with diet supplemented with 4% turmeric rhizome. Values represent mean+standard deviation (n ¼ 6). Values with the same superscript number on the same row are not significantly (p < 0.05) different.
level was significantly (p < 0.05) reduced in gentamycin-administered (control) rats (77.2 mg/dl), when compared to the normal rats (67.7 mg/dl). However, the urea level was significantly (p < 0.05) reduced in gentamycin-administered rats fed diets supplemented with ginger and turmeric rhizome (2% and 4%). Furthermore, the plasma BUN level significantly (p < 0.05) increased in gentamycin-administered (control) rats (36.1 mg/dl) when compared with the normal rats (31.6 mg/dl). However, BUN level was significantly (p < 0.05) reduced in gentamycin-administered rats fed diets supplemented with ginger and turmeric rhizome (2% and 4%) except those fed with 4% dietary inclusion of turmeric (34.1 mg/dl). The results show that plasma uric acid level significantly (p < 0.05) increases in gentamycin-administered (control) rats (7.2 mg/dl) when compared with the normal rats (5.4 mg/dl). However, the plasma uric acid level was significantly (p < 0.05) restored in gentamycin-administered rats fed diets supplemented with ginger and turmeric rhizome (2% and 4%). In addition, the results of the kidney malondialdehyde (MDA) and GSH levels are presented in Table 3. The results revealed that gentamycin administration caused a significant (p < 0.05) increase in the kidney MDA content of the control rats (3.5 mmol/g protein) with respect to the normal rats (2.4 mmol/g protein). However, this gentamycin-induced elevated kidney MDA content was significantly lowered in the groups fed diets supplemented with ginger and turmeric rhizomes (2% and 4% inclusions). Furthermore, Table 3 also depicts the effects of dietary inclusion of ginger and turmeric (2% and 4%) on the status of kidney antioxidant enzymes (CAT, GST, SOD and GPx). The results revealed significant (p < 0.05) reductions in the kidney activities of the enzymes following gentamycin administration compared with normal rats. However, there were significant (p < 0.05) improvements in the kidney antioxidant enzymes’ activities in the groups fed diets supplemented with ginger and turmeric rhizome (2% and 4%), with the exception of kidney SOD activities 6 Downloaded from nah.sagepub.com at GEORGIAN COURT UNIV on October 20, 2014
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that showed no significant (p > 0.05) increase in activity compared to the control rats, for groups fed diets supplement with either 2% or 4% ginger rhizome.
Discussion The observed percentage weight gain in rats fed diets supplemented with the spices (except 2% turmeric inclusion) compared to the control group could be due to the acceptability of the diets across the treatment groups. However, the ability of turmeric to induce appetite is well reported (Ammon and Wahl, 2001). Gentamycin has been shown to cause loss of appetite, plus increased catabolism characteristic of gentamycin-induced acute renal failure which results in acidosis accompanied by anorexia (Erdem et al., 2000). Consequently, oral food intake may reduce which may account for weight loss (Erdem et al., 2000). This is consistent with the findings of this study where significant weight loss was recorded in the control group. Previous studies have implicated oxidative stress in acute (Himmelfarb et al., 2004) and chronic renal failures (Annuk et al., 2001). Gentamycin is an antibiotic widely used mainly against gram-negative bacteria infections but which has been limited by its nephrotoxicity (Martinez-Salgado et al., 2007) with oxidative stress due to free radical generation being the major mechanism of its nephrotoxicity (Dhanarajan et al., 2006). Ginger rhizome has been previously reported to exhibit strong antioxidant properties which is linked to its rich phenolic content (Shirin and Jamuna, 2010). Similarly, curcumin, the major phytochemical of turmeric, has been shown to be a potent anti-inflammatory, anticancer, and most especially, antioxidant agent (Chan et al., 2009; Somasundaram et al., 2002). From the results obtained, gentamycin administration led to elevation of all the renal damage biomarkers; plasma creatinine, plasma urea, BUN and plasma uric acid (Table 2). This supports the claim that gentamycin induces renal damage in rats, and is characterized by an increase in plasma creatinine, urea, uric acid and BUN (Lakshmi and Sudhakar, 2010). The increase in plasma creatinine shows a depletion in glomerular filtration rate associated with an increase in plasma urea, uric acid and BUN levels as a result of significant renal parenchyma injury (Erdem et al., 2000). However, pretreatment of rats with diets supplemented with ginger and turmeric rhizome powder (2% and 4%) significantly improved all renal damage biomarkers, thus preventing the rats from being affected by the nephrotoxic effect of gentamycin. Interestingly, 4% ginger and 2% turmeric dietary inclusion significantly reduced the plasma uric acid levels far below the levels seen in normal rats showing their high nephroprotective potencies. Turmeric has been previously shown to have nephroprotective properties. Tirkey et al. (2005) showed that curcumin from turmeric significantly and dose-dependently improves cyclosporine-induced renal damage in rats by improving creatinine clearance and lowering the increase in serum creatinine and BUN. Gentamycin-associated toxicity has been shown to be responsible for impairment of the antioxidant system in the kidney (Ahmad, 1997) which supports the claim that reactive oxygen species (ROS) play key role in gentamycin-induced renal damage (Martinez-Salgado, 2007). Hydrogen peroxide has been shown to be one of the ROS induced by gentamycin in the renal mitochondrial cortex (Walker and Shah, 1988) and has been attributed to the increase in the generation of superoxide anions (Walker and Shah, 1988). Superoxide anions and hydrogen peroxide interact in a Fe2þ-catalyzed redox reaction to generate hydroxyl radical (Shah and Walker, 2002). These unstable 7 Downloaded from nah.sagepub.com at GEORGIAN COURT UNIV on October 20, 2014
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radicals can damage cells by starting chemical chain reactions such as lipid peroxidation, DNA and protein oxidation (Helmut, 1997). From this study, there was increased lipid peroxidation in gentamycin-induced control rats as reflected by the significant increase in the MDA content of kidney tissue (Table 3). The administration of diets supplemented with ginger and turmeric rhizome powder (2% and 4%) to the rats however, significantly reduced the MDA content; thus preventing lipid peroxidation. This is in agreement with the study of Tarasub et al. (2011) who showed that curcumin ameliorates lipid peroxidation in cadmium-induced nephrotoxicity in rats as well as that of Verma and Mathuria (2008) who reported that curcumin suppresses aflatoxin-induced lipid peroxidation in the liver and kidney in vitro in a dose-dependent manner. Similarly, Ahmed et al. (2000) have reported the antioxidant ability of ginger by decreasing lipid peroxidation, increasing GSH level and maintaining normal level of antioxidant enzymes. Furthermore, this study revealed that gentamycin-induced nephrotoxicity led to significant decrease in the kidney antioxidant enzymes; CAT, GST, GPx and SOD (Table 3). However, there was significant restoration of these enzymes in gentamycin-treated rats pretreated with diets supplemented with ginger and turmeric rhizome powder (2% and 4%). The diet supplemented with ginger rhizome powder (2% and 4%) gave an interesting result by restoring CAT activity to levels higher than those seen in normal rats in a dose-dependent fashion. Similarly, diet supplemented with turmeric rhizome powder (2% and 4%) restored the GPx activity levels to those higher than seen in the normal rats. However, there seems to be no significant difference in GSH content in the different treatment groups. Chen et al. (2008) reported the antioxidant properties of methanolic extracts of several ginger species. Also, several studies have reported the antioxidant properties of turmeric through its radical scavenging abilities (Ruby et al., 2005; Sharmas et al., 2006; Sreejayan and Rao, 1997). High Pressure Liquid Chromatography coupled to a diode array detector (HPLCDAD) analysis (unpublished data) on ginger and turmeric in our laboratory have revealed the presence of gallic acid, catechin, caffeic acid, (-)epigallocatechin, rutin, quercetin and kaempferol as the dominant phenolic constituents of ginger while, gallic acid, caffeic acid, quercetin and curcumin being the dominant phenolic constituents of turmeric. Phenolic compound are potent antioxidant compounds with strong free radical scavenging ability. Furthermore, biological activities of phenolic compounds have been linked to their antioxidant properties and these could be responsible for their involvement in the healing process of tissues subjected to oxidative stress. Thus, the observed nephroprotective effect of both ginger and turmeric observed in this study could be a function of their constituent phenolic phytochemicals.
Conclusion Dietary inclusion of ginger and turmeric (2% and 4%) can ameliorate renal damage which was evidenced in the ability of ginger and turmeric to reduce kidney oxidative stress, boost renal antioxidant status and inhibit lipid peroxidation in the kidney. Furthermore, the nephroprotetive effects of these spices were demonstrated by their ability to significantly improve kidney function by the restoration of kidney function indices. Therefore, dietary inclusion of ginger and turmeric rhizome could be a practical and cheap means of management of nephrotoxicity and acute renal damage. 8 Downloaded from nah.sagepub.com at GEORGIAN COURT UNIV on October 20, 2014
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Funding This research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors. Conflicts of Interest None declared. References Abdullahi M (2011) Biopotency role of culinary spices and herbs and their chemical constituents in health and commonly used spices in Nigerian dishes and snacks. African Journal of Food Science 5(3): 111–124. Adefegha SA and Oboh G (2012) Inhibition of key enzymes linked to type 2 diabetes and sodium nitroprusside-induced lipid peroxidation in rat pancreas by water extractable phytochemicals from some tropical spices. Pharmaceutical Biology 50(7): 857–865. Ahmad RSS (1997) Biochemical studies on combined effect of garlic (Allium sativum Linn.) and ginger (Zingiber officinale Rosc.) in albino rats. Indian Journal of Experimental Biology 35(8): 841–843. Ali G, Hawa ZEJ and Asmah R (2010) Synthesis of phenolics and flavonoids in ginger (Zingiber officinale Roscoe) and their effects on photosynthesis rate. International Journal of Molecular Sciences 11(11): 4539–4555. Alia M, Horcajo C, Bravo L, et al. (2003) Effect of grape antioxidant dietary fiber on the total antioxidant capacity and the activity of liver antioxidant enzymes in rats. Nurtition Research 23(9): 1251–1267. Ammon P and Wahl A (2001) Pharmacology of Curcuma longa. Planta Medica 57(1): 1–7. Annuk M, Zilmer M, Lind L, et al. (2001) Oxidative stress and endothelial function in chronic renal failure. Journal of the American Society of Nephrology 12(12): 2747–2751. Belle NAV, Dalmolin GD, Fonini G, et al. (2004) Polyamines reduces lipid peroxidation induced by different prooxidant agents. Brain Research 1008(2): 245–251. Biswas S, Moclure D and Jimenez A (2005) Curcumin induces glutathione bios-synthesis and inhibits NF kappa B activation and interleukin 8 release. Antioxidant and Redox Signalling 7(1-2): 32–41. Bushra HE and Effat MEA (2007) The protective effect of curcumin against gentamycin-induced renal dysfunction and oxidative stress in male albino rats. Egyptian Journal of Hospital Medicine 29: 546–556. Chan EWC, Lim YY, Wong SK, et al. (2009) Effects of different drying methods on the antioxidant properties of leaves and tea of ginger species. Food Chemistry 113(1): 166–172. Chen IN, Chang CC, Ng CC, et al. (2008) Antioxidant and antimicrobial activity of Zingiberaceae plants in Taiwan. Plant Food for Human Nutrition 63(1): 15–20. Dhanarajan R, Abraham P and Isaac B (2006) Protective effect of ebselen, a selenoorganic drug, against gentamicin-induced renal damage in rats. Basic and Clinical Pharmacology and Toxicology 99(3): 267–272. Ellman GL (1959) Tissue sulfhydryl groups. Archives of Biochemistry and Biophysics 82(1): 70–77. Erdem A, Gondogan NU, Usubatan A, et al. (2000) The protective effect of taurine against gentamicin-induced acute tubular necrosis in rats. Nephrology Dialysis Transplantation 15(8): 1175–1182. Habig WH, Pabst MJ and Jakoby WB (1974) Glutathione s-transferases: The first enzymatic step in mercapturic acid and formation. Journal of Biological Chemistry 249(22): 7130–7139. Hatcher H, Planalp R, Cho J, et al. (2008) Curcumin: From ancient medicine to current clinical trials. Cellular and Molecular Life Sciences 65(11): 1631–1652. 9 Downloaded from nah.sagepub.com at GEORGIAN COURT UNIV on October 20, 2014
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Helmut S (1997) Oxidative stress: Oxidation and antioxidants. Experimental Physiology 82(2): 291–295. Himmelfarb J, Mcmonagle E, Freedman S, et al. (2004) Oxidative stress is increased in critically ill patients with acute renal failure. Journal of the American Society of Nephrology 15(9): 2449–2456. Jurenka JS (2009) Anti-inflammatory properties of curcumin, major constituent of Curcuma longa: A review of preclinical and clinical research. Alternative Medicine Review 14(2): 141–153. Lakshmi BVS and Sudhakar M (2010) Protective effect of Zingiber officinale on gentamicininduced nephrotoxicity in rats. International Journal of Pharmacology 6(1): 58–62. Lowry OH, Rosebrough NJ, Farr AL, et al. (1951) Protein measurement with the Folinphenol reagent. Journal of Biological Chemistry 193: 265–275. Martı´nez-Salgado C, Lo´pez-Herna´ndez FJ and Lo´pez-Novoa JM (2007) Glomerular nephrotoxicity of aminoglycosides. Toxicology and Applied Pharmacology 223(1): 86–98. Moulds RFW and Jeyasingham MS (2010) Gentamicin, a great way to start. Australian Prescriber 33(5): 134–135. Oboh G, Akomolafe TL and Adetuyi AO (2010) Inhibition of cyclophosphamide induced oxidative stress in brain by dietary inclusion of red dye extracts from sorghum (Sorghum bicolor) stem. Journal of Medicinal Food 13(5): 1075–1080. Ohkawa H, Ohishi N and Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Analytical Biochemistry 95(2): 351–358. Rotruck JT, Pope AL, Ganther HE, et al. (1973) Selenium: Biochemical role as a component of glutathione peroxidase. Science 179(4073): 588–590. Ruby AJ, Kuttan G, Babu KD, et al. (2005) Anti-tumour and antioxidant activity of natural curcuminoids. Cancer Letters 94(1): 79–83. Shah V and Walker D (2002) Reactive oxygen metabolites in toxic acute renal failure. Renal Failure 14(3): 363–370. Shaibu OB and Chika A (2009) Dose-dependent amelioration of gentamicin-induced nephrotoxicity in adult Swiss albino rats by vitamin B-complex - a preliminary study. Tropical Journal of Pharmaceutical Research 8(2): 111–116. Sharmas S, Kulkarn K and Chopra K (2006) Curcumin, the active principle of turmeric (Curcuma longa), ameliorates diabetic nephropathy in rats. Clinical and Experimental Pharmacology and Physiology 33(10): 940–945. Shirin APR and Jamuna P (2010) Chemical composition and antioxidant properties of ginger root (Zingiber officinale). Journal of Medicinal Plants Research 4(24): 2674–2679. Sinha AK (1972) Colorimetric assay of catalase. Analytical Biochemistry 47(2): 389–394. Somasundaram S, Edmund NA, Moore DT, et al. (2002) Dietary curcumin inhibits chemotherapy-induced apoptosis in models of human breast cancer. Cancer Research 62(13): 3868–3875. Sreejayan I and Rao N (1997) Nitric oxide scavenging by curcuminoids. Journal of Pharmacy and Pharmacology 49(1): 105–107. Tarasub N, Tarasub C and Devakul NW (2011) Protective effect of curcumin on cadmium-induced nehprotoxicity in rats. Journal of Environmental Chemistry and Ecotoxicology 3(2): 17–24. Tirkey N, Kaur G, Vij G, et al. (2005) Curcumin, a diferuloylmethane, attenuates cyclosporineinduced renal dysfunction and oxidative stress in rat kidneys. BMC Pharmacology 5:15. Verma RJ and Mathuria N (2008) Curcumin ameliorates aflatoxin-induced lipid-peroxidation in liver and kidney of mice. Acta Poloniae Pharmaceutica 65(2): 195–202. Walker PD and Shah SV (1988) Evidence suggesting a role for hydroxyl radical in gentamycin-induced acute renal failure in rats. Journal of Clinical Investigation 81(2): 334–341. 10
Attenuation of Gentamycin-induced nephrotoxicity in rats by dietary inclusion of ginger ( Zingiber officinale) and turmeric (Curcuma longa) rhizomes Adedayo O Ademiluyi, Ganiyu Oboh, Opeyemi B Ogunsuyi and Ayodele J. Akinyemi Nutrition and Health published online 5 November 2013 DOI: 10.1177/0260106013506668 The online version of this article can be found at: http://nah.sagepub.com/content/early/2013/10/17/0260106013506668 A more recent version of this article was published on - Mar 11, 2014
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Article
Attenuation of gentamycin-induced nephrotoxicity in rats by dietary inclusion of ginger (Zingiber officinale) and turmeric (Curcuma longa) rhizomes
Nutrition and Health 1-10 ª The Author(s) 2013 Reprints and permission: sagepub.co.uk/journalsPermissions.nav DOI: 10.1177/0260106013506668 nah.sagepub.com
Adedayo O Ademiluyi1, Ganiyu Oboh1, Opeyemi B Ogunsuyi1, and Ayodele J Akinyemi1,2
Abstract This study sought to investigate the modulatory effects of dietary inclusion of ginger (Zingiber officinale) and turmeric (Curcuma longa) rhizomes on antioxidant status and renal damage induced by gentamycin in rats. Renal damage was induced in albino rats pretreated with dietary inclusion of ginger and turmeric (2% and 4%) by intraperitoneal (i.p.) administration of gentamycin (100 mg/kg body weight) for three days. Assays for renal damage biomarkers (plasma creatinine, plasma urea, blood urea nitrogen and plasma uric acid), malondialdehyde (MDA) content and reduced glutathione (GSH) content as well as renal antioxidant enzymes (catalase, glutathione-Stransferase (GST), glutathione peroxidase (GPx) and superoxide dismutase (SOD)) were carried out. The study revealed significant (p < 0.05) increases in renal damage biomarkers following gentamycin administration with severe alteration in kidney antioxidant status. However, pretreatment with ginger and turmeric rhizome (2% and 4%) prior to gentamycin administration significantly (p < 0.05) protected the kidney and attenuated oxidative stress by modulating renal damage and antioxidant
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Functional Foods and Nutraceuticals Unit, Federal University of Technology, Akure, Nigeria Department of Biochemistry, Afe Babalola University, Ado-Ekiti, Nigeria
Corresponding author: Adedayo O Ademiluyi, Functional Foods and Nutraceuticals Unit, Department of Biochemistry, Federal University of Technology, Akure P.M.B. 704, Akure, 340001, Nigeria. Email: [email protected]
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indices. This finding therefore suggests that dietary inclusion of ginger and turmeric rhizomes may protect against gentamycin-induced nephrotoxicity and oxidative stress. Keywords Ginger, Zingiber officinale, turmeric, Curcuma longa, gentamycin, nephrotoxicity, oxidative stress
Introduction The kidneys are vital organs essential for excretion of metabolic wastes as well as maintaining chemical homeostasis among other functions. Many research have linked oxidative stress as a potential cause of different forms of renal damage or nephrotoxicity. Studies have shown that increase in oxidative stress due to free radical generation is a likely result of inflammatory responses associated with patients suffering from acute renal failure (Himmelfarb et al., 2004), while accumulated evidences give credence to the fact that there is elevated level of oxidative stress in patients with chronic renal failure which is often accompanied by proportional endothelial cell damage (Annuk et al., 2001). Considering the vital physiological functions of the kidney and the unique role played by free radical-induced oxidative stress in different forms of acute and chronic renal damage, it is important to explore ways by which oxidative stress may be ameliorated. Xenobiotic interaction with renal tubular cells is one way of generating free radicals in the kidney. Gentamycin is a bacteriocidal antibiotic (Martı´nez-Salgado et al., 2007) used mainly against gram-negative bacteria (Moulds and Jeyasingham, 2010). However, this drug has some side effects with nephrotoxicity and ototoxicity being most studied (Martı´nez-Salgado et al., 2007). Nephrotoxicity resulting from gentamycin has been established at a dose above 2 mg/ml in humans (Shaibu and Chika, 2009). Studies have suspected that gentamycin induces free radical generation in renal tubular cells to elicit nephrotoxicity (Martı´nez-Salgado et al., 2007). Gentamycin is concentrated in the renal proximal tubular cells where it induces generation of superoxide anions, hydrogen peroxide and hydroxyl radicals. However, a lack of detailed understanding of the mechanism of toxicity of gentamycin has made it difficult to control its toxicity (Shaibu and Chika, 2009). Nevertheless, the use of some synthetic antioxidant compounds/molecules to counteract the effects of free radical generation has been suggested as a viable management strategy, though their attendant side effects are a major drawback, hence the need for alternatives having little or no side effects, with plant sources being the most promising. Spices have been widely used as food adjunct and in folk medicine for several centuries (Adefegha and Oboh, 2012). Ginger (Zingiber officinale) rhizome is used widely as a spice for flavoring foods and has wide applications in folk medicine. Studies have shown its anti-inflammatory, hypoglycemic and hypolipidemic properties (Abdullahi, 2011; Ahmad, 1997). The antioxidant property of Z. officinale has been linked to its 2 Downloaded from nah.sagepub.com at GEORGIAN COURT UNIV on October 20, 2014
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volatile oil, polyphenol and flavonoid content (Ali et al., 2010; Lakshmi and Sudhakar, 2010). Furthermore, turmeric (Curcuma longa) which is another rhizomatous plant belonging to the ginger family has been employed as a dye source and food colorant due to its color, and it is also used as a food flavoring agent. Curcuminoids are the major phytochemicals of turmeric which are responsible for the characteristic yellow color (Jurenka, 2009). Curcumin, one of the predominant curcuminoids and a flavonoid, has been investigated for anti-inflammatory and antioxidant properties (Hatcher et al., 2008). Curcumin has also been studied to ‘mop up’ superoxide anions, peroxynitrite radicals and singlet oxygen (Biswas et al., 2005). The ability to derive cheap dietary sources of antioxidants in ginger and turmeric offers a potential dual benefit for their use as culinary spices and to ameliorate oxidative stress. Thus, antioxidant sources with no known toxicological side effects (at safe dosage) offer a good way to manage nephrotoxicity generated by the use of gentamycin.
Materials and method Materials Fresh rhizomes of ginger and turmeric were purchased from Oja Oba market in Akure, Ondo State Nigeria. The authentication of the samples was done in the Department of Crop, Soil and Pest Management, Federal University of Technology, Akure, Nigeria. These samples were washed, chopped and sun-dried to dryness after which they were ground into fine powder. All chemicals and reagents used were of analytical grade and distilled water was used. All the kits used for bioassays were sourced from Randox Laboratories Ltd., Crumlin, Co. Antrim, UK. Gentamycin was sourced as Enagent Gentamycin Sulphate Injection (Zhejang, JinlingTiafeng Pharmaceutical Factory, Chezhan Road, Huzhou City, Zhejiiang, China). Diet ingredients were purchased from VITAL Feeds, Jos, Nigeria Ltd.
Animals Male albino rats weighing 97–121 g used for this experiment were purchased from the breeding colony of a private farm in Akure, Ondo State, Nigeria. The rats were maintained at 25oC on a 12 h light/dark cycle with unrestricted access to food and water. They were acclimatized under these conditions for two weeks prior to the commencement of the experiments. This study received the approval of the institution’s Ethical Committee on the use of laboratory animals and the animals were handled in accordance with National Institute of Health (NIH) guide for the care and use of laboratory animals.
Experimental design and induction of nephrotoxicity After acclimatization, the rats were randomly divided into six groups of six animals each. Group I and Group II were fed a basal diet (50% skimmed milk, 36% corn starch, 10% groundnut oil and 4% mineral & vitamin premix) following a slightly modified method of Oboh et al. (2010), Groups III and IV were fed basal diet supplemented with 2% and 4% ginger rhizome respectively and groups V and VI were fed basal diet supplemented 3 Downloaded from nah.sagepub.com at GEORGIAN COURT UNIV on October 20, 2014
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with 2% and 4% turmeric respectively for 27 days prior to gentamycin (100 mg/kg body weight i.p.) administration (Bushra and Effat, 2007) which lasted for three days and the experiment was terminated after 30 days. The rats were subject to an overnight fast after which they were decapitated by cervical dislocation. The blood was rapidly collected into separate Ethylene diaminetetraacetic acid (EDTA) bottles by direct heart puncture and centrifuged at 800g for 10 min to separate the plasma. Similarly, the kidneys were isolated, rinsed in cold saline (0.9% NaCl) and homogenized in phosphate buffer (pH 6.9). The homogenates were centrifuged at 5000g for 10 min to obtain the clear supernatant. The clear supernatant obtained was used for various biochemical assays (Belle et al., 2004).
Analytical procedures The assay for the renal damage biomarkers; plasma creatinine, plasma urea and plasma uric acid were carried out according to the manufacturer’s procedure of the commercially available kits (Randox Laboratories UK). Blood urea nitrogen (BUN) was subsequently calculated from the plasma urea concentration. Lipid peroxidation was determined as reported by Ohkawa et al. (1979) with slight modification; catalase (CAT) activity was assayed as reported by Sinha (1972); glutathione-S-transferase (GST) activity was assayed according to Habig et al., (1974); glutathione peroxidase (GPx) activity was determined by the method of Rotruck et al. (1973); superoxide dismutase (SOD) activity was determined by the method of Alia et al. (2003); reduced glutathione (GSH) content was determined by the modified method of Ellman (1959) and total protein was determined according to Lowry et al. (1951).
Statistical analysis The results of replicate experiments were represented as mean+standard deviation (SD). Statistical analyses were done using statistical program for social science (SPSS) 16.0 (SPSS. Inc. Chicago, Illinois, USA). One way analysis of variance (ANOVA) was carried out followed by Duncan’s test for the post-hoc treatment. A value of p < 0.05 was considered to be statistically significant.
Results The percentage average weight gain/loss of the rats in each treatment groups is presented in Table 1. Weights were recorded before feeding the rats with supplemented diets (Day 1) and 27 days after feeding them with the supplemented diets. The result revealed a significant (p < 0.05) percentage weight increase across the six groups of experimental rats after feeding on the formulated diets for 27 days. However, three days after gentamycin (100 mg/kg body weight i.p.) administration (Day 30) to the rats, a significant weight decrease was recorded in the control rats. Nevertheless, the groups fed diets supplemented with ginger and turmeric rhizome (2% and 4%) recorded significant weight gain compared to the control rats (except those with 2% turmeric inclusion). 4 Downloaded from nah.sagepub.com at GEORGIAN COURT UNIV on October 20, 2014
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Table 1. Effect of dietary inclusion of ginger and turmeric rhizome on average weight gain/loss in gentamycin-administered rats. Weight (g/rat) Groups I II III IV V VI
Day 1
Day 27
165.8+39.6 150.8+52.2 84.9+70.0 91.3+56.4 143.4+60.9 107.4+37.5
170.6+29.9 167.3+31.0 90.3+31.0 95.4+46.1 158.7+35.2 138.7+2.6
Weight (g/rat) Weight gain/loss (%) a
2.9 10.9c 6.3b 4.5b 10.7c 29.1d
Day 30
Weight gain/loss (%)
192.9+40.0 148.9+29.6 95.3+51.8 99.7+44.2 147.2+37.3 157.6+18.9
13.1c –11.0a 5.5b 4.5b –7.6ab 13.7c
I: normal rats fed with basal diet; II: control rats (gentamycin-administered rats fed with basal diet); III: gentamycin-administered rats fed with diet supplemented with 2% ginger rhizome; IV: gentamycinadministered rats fed with diet supplemented with 4% ginger rhizome; V: gentamycin-administered rats fed with diet supplemented with 2% turmeric; VI: gentamycin-administered rats fed with diet supplemented with 4% turmeric rhizome. Values represent mean+standard deviation (n ¼ 6). Values with the same superscript number on the same row are not significantly (p < 0.05) different.
Table 2. Effect of dietary inclusion of ginger and turmeric rhizome on renal damage biomarkers in gentamycin-administered rats. Plasma concentration of renal damage biomarkers (mg/dl) Groups I II III IV V VI
Creatinine
Urea
BUN
Uric acid
0.25+0.05a 0.36+0.13c 0.34+0.01bc 0.32+0.04bc 0.31+0.08abc 0.29+0.05ab
67.7+6.4a 77.2+2.0c 72.1+6.9b 66.9+6.2a 71.1+2.5b 73.1+2.7c
31.6+3.0a 36.1+0.9c 33.7+3.2b 31.3+2.9a 33.2+1.2b 34.1+1.3c
5.4+1.0bc 7.2+0.8d 6.0+0.5cd 2.8+1.3a 3.9+0.1b 5.8+0.8cd
I: normal rats fed with basal diet; II: control rats (gentamycin-administered rats fed with basal diet); III: gentamycin-administered rats fed with diet supplemented with 2% ginger rhizome; IV: gentamycinadministered rats fed with diet supplemented with 4% ginger rhizome; V: gentamycin-administered rats fed with diet supplemented with 2% turmeric; VI: gentamycin-administered rats fed with diet supplemented with 4% turmeric rhizome. Values represent mean+standard deviation (n ¼ 6). Values with the same superscript number on the same row are not significantly (p < 0.05) different
Table 2 shows the effect of the dietary inclusion of ginger and turmeric rhizome on the renal damage biomarkers (plasma creatinine, urea, BUN and uric acid) in normal rats and gentamycin-administered rats. The result shows that the plasma level of creatinine significantly (p < 0.05) increases in gentamycin-administered (control) rats (0.36 mg/ dl) when compared with the normal rats (0.25 mg/dl). However, the plasma creatinine level was significantly (p < 0.05) restored in gentamycin-administered rats fed diets supplemented with ginger and turmeric rhizome (2% and 4%). Similarly, the plasma urea 5 Downloaded from nah.sagepub.com at GEORGIAN COURT UNIV on October 20, 2014
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Table 3. Effect of dietary inclusion of ginger and turmeric rhizome on tissue enzymatic and nonenzymatic antioxidant indices in gentamycin-administered rats. Groups I II III IV V VI
Catalase U/g protein
GST U/g protein
5.4+0.4bc 4.4+0.2a 5.9+0.5c 7.0+0.3d 5.0+0.7b 5.0+0.3ab
3.0+0.6c 2.0+0.4ab 1.7+0.2a 2.9+0.6bc 2.6+0.5bc 2.8+0.5bc
GPx U/g protein
SOD U/g protein
1.1+0.032a 11.3+3.6c 0.9+ 0.23c 6.9+1.8abc 0.9+ 0.27b 6.2+1.0ab 1.2+ 0.31a 4.7+1.8a 1.3+ 0.18b 9.2+3.0c 1.7+ 0.23c 8.2+1.7bc
MDA mmol/g protein
GSH mg/g protein
2.4+0.9a 3.5+0.7b 2.5+0.1a 2.4+0.5a 2.3+0.9a 2.2+0.1a
13.8+2.3a 10.1+1.6a 12.8+2.0a 13.1+3.1a 12.9+1.5a 13.0+2.0a
GPx: glutathione peroxidase; GSH: reduced glutathione; GST: glutathione-S-transferase; MDA: malondialdehyde; SOD: superoxide dismutase. I: normal rats fed with basal diet; II: control rats (gentamycin-administered rats fed with basal diet); III: gentamycin-administered rats fed with diet supplemented with 2% ginger rhizome; IV: gentamycinadministered rats fed with diet supplemented with 4% ginger rhizome; V: gentamycin-administered rats fed with diet supplemented with 2% turmeric; VI: gentamycin-administered rats fed with diet supplemented with 4% turmeric rhizome. Values represent mean+standard deviation (n ¼ 6). Values with the same superscript number on the same row are not significantly (p < 0.05) different.
level was significantly (p < 0.05) reduced in gentamycin-administered (control) rats (77.2 mg/dl), when compared to the normal rats (67.7 mg/dl). However, the urea level was significantly (p < 0.05) reduced in gentamycin-administered rats fed diets supplemented with ginger and turmeric rhizome (2% and 4%). Furthermore, the plasma BUN level significantly (p < 0.05) increased in gentamycin-administered (control) rats (36.1 mg/dl) when compared with the normal rats (31.6 mg/dl). However, BUN level was significantly (p < 0.05) reduced in gentamycin-administered rats fed diets supplemented with ginger and turmeric rhizome (2% and 4%) except those fed with 4% dietary inclusion of turmeric (34.1 mg/dl). The results show that plasma uric acid level significantly (p < 0.05) increases in gentamycin-administered (control) rats (7.2 mg/dl) when compared with the normal rats (5.4 mg/dl). However, the plasma uric acid level was significantly (p < 0.05) restored in gentamycin-administered rats fed diets supplemented with ginger and turmeric rhizome (2% and 4%). In addition, the results of the kidney malondialdehyde (MDA) and GSH levels are presented in Table 3. The results revealed that gentamycin administration caused a significant (p < 0.05) increase in the kidney MDA content of the control rats (3.5 mmol/g protein) with respect to the normal rats (2.4 mmol/g protein). However, this gentamycin-induced elevated kidney MDA content was significantly lowered in the groups fed diets supplemented with ginger and turmeric rhizomes (2% and 4% inclusions). Furthermore, Table 3 also depicts the effects of dietary inclusion of ginger and turmeric (2% and 4%) on the status of kidney antioxidant enzymes (CAT, GST, SOD and GPx). The results revealed significant (p < 0.05) reductions in the kidney activities of the enzymes following gentamycin administration compared with normal rats. However, there were significant (p < 0.05) improvements in the kidney antioxidant enzymes’ activities in the groups fed diets supplemented with ginger and turmeric rhizome (2% and 4%), with the exception of kidney SOD activities 6 Downloaded from nah.sagepub.com at GEORGIAN COURT UNIV on October 20, 2014
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that showed no significant (p > 0.05) increase in activity compared to the control rats, for groups fed diets supplement with either 2% or 4% ginger rhizome.
Discussion The observed percentage weight gain in rats fed diets supplemented with the spices (except 2% turmeric inclusion) compared to the control group could be due to the acceptability of the diets across the treatment groups. However, the ability of turmeric to induce appetite is well reported (Ammon and Wahl, 2001). Gentamycin has been shown to cause loss of appetite, plus increased catabolism characteristic of gentamycin-induced acute renal failure which results in acidosis accompanied by anorexia (Erdem et al., 2000). Consequently, oral food intake may reduce which may account for weight loss (Erdem et al., 2000). This is consistent with the findings of this study where significant weight loss was recorded in the control group. Previous studies have implicated oxidative stress in acute (Himmelfarb et al., 2004) and chronic renal failures (Annuk et al., 2001). Gentamycin is an antibiotic widely used mainly against gram-negative bacteria infections but which has been limited by its nephrotoxicity (Martinez-Salgado et al., 2007) with oxidative stress due to free radical generation being the major mechanism of its nephrotoxicity (Dhanarajan et al., 2006). Ginger rhizome has been previously reported to exhibit strong antioxidant properties which is linked to its rich phenolic content (Shirin and Jamuna, 2010). Similarly, curcumin, the major phytochemical of turmeric, has been shown to be a potent anti-inflammatory, anticancer, and most especially, antioxidant agent (Chan et al., 2009; Somasundaram et al., 2002). From the results obtained, gentamycin administration led to elevation of all the renal damage biomarkers; plasma creatinine, plasma urea, BUN and plasma uric acid (Table 2). This supports the claim that gentamycin induces renal damage in rats, and is characterized by an increase in plasma creatinine, urea, uric acid and BUN (Lakshmi and Sudhakar, 2010). The increase in plasma creatinine shows a depletion in glomerular filtration rate associated with an increase in plasma urea, uric acid and BUN levels as a result of significant renal parenchyma injury (Erdem et al., 2000). However, pretreatment of rats with diets supplemented with ginger and turmeric rhizome powder (2% and 4%) significantly improved all renal damage biomarkers, thus preventing the rats from being affected by the nephrotoxic effect of gentamycin. Interestingly, 4% ginger and 2% turmeric dietary inclusion significantly reduced the plasma uric acid levels far below the levels seen in normal rats showing their high nephroprotective potencies. Turmeric has been previously shown to have nephroprotective properties. Tirkey et al. (2005) showed that curcumin from turmeric significantly and dose-dependently improves cyclosporine-induced renal damage in rats by improving creatinine clearance and lowering the increase in serum creatinine and BUN. Gentamycin-associated toxicity has been shown to be responsible for impairment of the antioxidant system in the kidney (Ahmad, 1997) which supports the claim that reactive oxygen species (ROS) play key role in gentamycin-induced renal damage (Martinez-Salgado, 2007). Hydrogen peroxide has been shown to be one of the ROS induced by gentamycin in the renal mitochondrial cortex (Walker and Shah, 1988) and has been attributed to the increase in the generation of superoxide anions (Walker and Shah, 1988). Superoxide anions and hydrogen peroxide interact in a Fe2þ-catalyzed redox reaction to generate hydroxyl radical (Shah and Walker, 2002). These unstable 7 Downloaded from nah.sagepub.com at GEORGIAN COURT UNIV on October 20, 2014
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radicals can damage cells by starting chemical chain reactions such as lipid peroxidation, DNA and protein oxidation (Helmut, 1997). From this study, there was increased lipid peroxidation in gentamycin-induced control rats as reflected by the significant increase in the MDA content of kidney tissue (Table 3). The administration of diets supplemented with ginger and turmeric rhizome powder (2% and 4%) to the rats however, significantly reduced the MDA content; thus preventing lipid peroxidation. This is in agreement with the study of Tarasub et al. (2011) who showed that curcumin ameliorates lipid peroxidation in cadmium-induced nephrotoxicity in rats as well as that of Verma and Mathuria (2008) who reported that curcumin suppresses aflatoxin-induced lipid peroxidation in the liver and kidney in vitro in a dose-dependent manner. Similarly, Ahmed et al. (2000) have reported the antioxidant ability of ginger by decreasing lipid peroxidation, increasing GSH level and maintaining normal level of antioxidant enzymes. Furthermore, this study revealed that gentamycin-induced nephrotoxicity led to significant decrease in the kidney antioxidant enzymes; CAT, GST, GPx and SOD (Table 3). However, there was significant restoration of these enzymes in gentamycin-treated rats pretreated with diets supplemented with ginger and turmeric rhizome powder (2% and 4%). The diet supplemented with ginger rhizome powder (2% and 4%) gave an interesting result by restoring CAT activity to levels higher than those seen in normal rats in a dose-dependent fashion. Similarly, diet supplemented with turmeric rhizome powder (2% and 4%) restored the GPx activity levels to those higher than seen in the normal rats. However, there seems to be no significant difference in GSH content in the different treatment groups. Chen et al. (2008) reported the antioxidant properties of methanolic extracts of several ginger species. Also, several studies have reported the antioxidant properties of turmeric through its radical scavenging abilities (Ruby et al., 2005; Sharmas et al., 2006; Sreejayan and Rao, 1997). High Pressure Liquid Chromatography coupled to a diode array detector (HPLCDAD) analysis (unpublished data) on ginger and turmeric in our laboratory have revealed the presence of gallic acid, catechin, caffeic acid, (-)epigallocatechin, rutin, quercetin and kaempferol as the dominant phenolic constituents of ginger while, gallic acid, caffeic acid, quercetin and curcumin being the dominant phenolic constituents of turmeric. Phenolic compound are potent antioxidant compounds with strong free radical scavenging ability. Furthermore, biological activities of phenolic compounds have been linked to their antioxidant properties and these could be responsible for their involvement in the healing process of tissues subjected to oxidative stress. Thus, the observed nephroprotective effect of both ginger and turmeric observed in this study could be a function of their constituent phenolic phytochemicals.
Conclusion Dietary inclusion of ginger and turmeric (2% and 4%) can ameliorate renal damage which was evidenced in the ability of ginger and turmeric to reduce kidney oxidative stress, boost renal antioxidant status and inhibit lipid peroxidation in the kidney. Furthermore, the nephroprotetive effects of these spices were demonstrated by their ability to significantly improve kidney function by the restoration of kidney function indices. Therefore, dietary inclusion of ginger and turmeric rhizome could be a practical and cheap means of management of nephrotoxicity and acute renal damage. 8 Downloaded from nah.sagepub.com at GEORGIAN COURT UNIV on October 20, 2014
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Funding This research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors. Conflicts of Interest None declared. References Abdullahi M (2011) Biopotency role of culinary spices and herbs and their chemical constituents in health and commonly used spices in Nigerian dishes and snacks. African Journal of Food Science 5(3): 111–124. Adefegha SA and Oboh G (2012) Inhibition of key enzymes linked to type 2 diabetes and sodium nitroprusside-induced lipid peroxidation in rat pancreas by water extractable phytochemicals from some tropical spices. Pharmaceutical Biology 50(7): 857–865. Ahmad RSS (1997) Biochemical studies on combined effect of garlic (Allium sativum Linn.) and ginger (Zingiber officinale Rosc.) in albino rats. Indian Journal of Experimental Biology 35(8): 841–843. Ali G, Hawa ZEJ and Asmah R (2010) Synthesis of phenolics and flavonoids in ginger (Zingiber officinale Roscoe) and their effects on photosynthesis rate. International Journal of Molecular Sciences 11(11): 4539–4555. Alia M, Horcajo C, Bravo L, et al. (2003) Effect of grape antioxidant dietary fiber on the total antioxidant capacity and the activity of liver antioxidant enzymes in rats. Nurtition Research 23(9): 1251–1267. Ammon P and Wahl A (2001) Pharmacology of Curcuma longa. Planta Medica 57(1): 1–7. Annuk M, Zilmer M, Lind L, et al. (2001) Oxidative stress and endothelial function in chronic renal failure. Journal of the American Society of Nephrology 12(12): 2747–2751. Belle NAV, Dalmolin GD, Fonini G, et al. (2004) Polyamines reduces lipid peroxidation induced by different prooxidant agents. Brain Research 1008(2): 245–251. Biswas S, Moclure D and Jimenez A (2005) Curcumin induces glutathione bios-synthesis and inhibits NF kappa B activation and interleukin 8 release. Antioxidant and Redox Signalling 7(1-2): 32–41. Bushra HE and Effat MEA (2007) The protective effect of curcumin against gentamycin-induced renal dysfunction and oxidative stress in male albino rats. Egyptian Journal of Hospital Medicine 29: 546–556. Chan EWC, Lim YY, Wong SK, et al. (2009) Effects of different drying methods on the antioxidant properties of leaves and tea of ginger species. Food Chemistry 113(1): 166–172. Chen IN, Chang CC, Ng CC, et al. (2008) Antioxidant and antimicrobial activity of Zingiberaceae plants in Taiwan. Plant Food for Human Nutrition 63(1): 15–20. Dhanarajan R, Abraham P and Isaac B (2006) Protective effect of ebselen, a selenoorganic drug, against gentamicin-induced renal damage in rats. Basic and Clinical Pharmacology and Toxicology 99(3): 267–272. Ellman GL (1959) Tissue sulfhydryl groups. Archives of Biochemistry and Biophysics 82(1): 70–77. Erdem A, Gondogan NU, Usubatan A, et al. (2000) The protective effect of taurine against gentamicin-induced acute tubular necrosis in rats. Nephrology Dialysis Transplantation 15(8): 1175–1182. Habig WH, Pabst MJ and Jakoby WB (1974) Glutathione s-transferases: The first enzymatic step in mercapturic acid and formation. Journal of Biological Chemistry 249(22): 7130–7139. Hatcher H, Planalp R, Cho J, et al. (2008) Curcumin: From ancient medicine to current clinical trials. Cellular and Molecular Life Sciences 65(11): 1631–1652. 9 Downloaded from nah.sagepub.com at GEORGIAN COURT UNIV on October 20, 2014
10
Nutrition and Health
Helmut S (1997) Oxidative stress: Oxidation and antioxidants. Experimental Physiology 82(2): 291–295. Himmelfarb J, Mcmonagle E, Freedman S, et al. (2004) Oxidative stress is increased in critically ill patients with acute renal failure. Journal of the American Society of Nephrology 15(9): 2449–2456. Jurenka JS (2009) Anti-inflammatory properties of curcumin, major constituent of Curcuma longa: A review of preclinical and clinical research. Alternative Medicine Review 14(2): 141–153. Lakshmi BVS and Sudhakar M (2010) Protective effect of Zingiber officinale on gentamicininduced nephrotoxicity in rats. International Journal of Pharmacology 6(1): 58–62. Lowry OH, Rosebrough NJ, Farr AL, et al. (1951) Protein measurement with the Folinphenol reagent. Journal of Biological Chemistry 193: 265–275. Martı´nez-Salgado C, Lo´pez-Herna´ndez FJ and Lo´pez-Novoa JM (2007) Glomerular nephrotoxicity of aminoglycosides. Toxicology and Applied Pharmacology 223(1): 86–98. Moulds RFW and Jeyasingham MS (2010) Gentamicin, a great way to start. Australian Prescriber 33(5): 134–135. Oboh G, Akomolafe TL and Adetuyi AO (2010) Inhibition of cyclophosphamide induced oxidative stress in brain by dietary inclusion of red dye extracts from sorghum (Sorghum bicolor) stem. Journal of Medicinal Food 13(5): 1075–1080. Ohkawa H, Ohishi N and Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Analytical Biochemistry 95(2): 351–358. Rotruck JT, Pope AL, Ganther HE, et al. (1973) Selenium: Biochemical role as a component of glutathione peroxidase. Science 179(4073): 588–590. Ruby AJ, Kuttan G, Babu KD, et al. (2005) Anti-tumour and antioxidant activity of natural curcuminoids. Cancer Letters 94(1): 79–83. Shah V and Walker D (2002) Reactive oxygen metabolites in toxic acute renal failure. Renal Failure 14(3): 363–370. Shaibu OB and Chika A (2009) Dose-dependent amelioration of gentamicin-induced nephrotoxicity in adult Swiss albino rats by vitamin B-complex - a preliminary study. Tropical Journal of Pharmaceutical Research 8(2): 111–116. Sharmas S, Kulkarn K and Chopra K (2006) Curcumin, the active principle of turmeric (Curcuma longa), ameliorates diabetic nephropathy in rats. Clinical and Experimental Pharmacology and Physiology 33(10): 940–945. Shirin APR and Jamuna P (2010) Chemical composition and antioxidant properties of ginger root (Zingiber officinale). Journal of Medicinal Plants Research 4(24): 2674–2679. Sinha AK (1972) Colorimetric assay of catalase. Analytical Biochemistry 47(2): 389–394. Somasundaram S, Edmund NA, Moore DT, et al. (2002) Dietary curcumin inhibits chemotherapy-induced apoptosis in models of human breast cancer. Cancer Research 62(13): 3868–3875. Sreejayan I and Rao N (1997) Nitric oxide scavenging by curcuminoids. Journal of Pharmacy and Pharmacology 49(1): 105–107. Tarasub N, Tarasub C and Devakul NW (2011) Protective effect of curcumin on cadmium-induced nehprotoxicity in rats. Journal of Environmental Chemistry and Ecotoxicology 3(2): 17–24. Tirkey N, Kaur G, Vij G, et al. (2005) Curcumin, a diferuloylmethane, attenuates cyclosporineinduced renal dysfunction and oxidative stress in rat kidneys. BMC Pharmacology 5:15. Verma RJ and Mathuria N (2008) Curcumin ameliorates aflatoxin-induced lipid-peroxidation in liver and kidney of mice. Acta Poloniae Pharmaceutica 65(2): 195–202. Walker PD and Shah SV (1988) Evidence suggesting a role for hydroxyl radical in gentamycin-induced acute renal failure in rats. Journal of Clinical Investigation 81(2): 334–341. 10
