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ORIGINAL ARTICLE
Year : 2021  |  Volume : 9  |  Issue : 1  |  Page : 17-24

Effects of aqueous extract of Cyperus esculentus (tiger nut) on antioxidant status and hematological indices in the heart of cadmium-induced wistar rats


1 Department of Anatomy, School of Basic Medical Sciences, University of Benin, Benin City, Nigeria
2 Department of Medical Biochemistry, School of Basic Medical Sciences, University of Benin, Benin City, Nigeria
3 Department of Veterinary Physiology and Biochemistry, Faculty of Veterinary Medicine, University of Benin, Benin City, Nigeria

Date of Submission21-Aug-2020
Date of Decision27-Aug-2020
Date of Acceptance02-Sep-2020
Date of Web Publication20-May-2021

Correspondence Address:
Dr. Silvanus Olu Innih
Department of Anatomy, School of Basic Medical Sciences, University of Benin, Benin City
Nigeria
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/njecp.njecp_32_20

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  Abstract 


Background: In recent times, medicinal plants has been explored for their ameliorating effect on Cadmium induced organ toxicity in Wistar rats. Aims and Objectives: This study investigates the protective effects of aqueous extract of Cyperus esculentus (tiger nut) on cadmium-induced biochemical and histological alterations in heart of rats. Materials and Methods: Thirty male wistar rats were randomly divided into six groups (A-F). Blood samples were collected from the animals after treatment with cadmium (10mg/kg body weight) and varying doses (150, 300, 600mg/kg B.W) of aqueous extract of Cyperus esculentus for 60 days for evaluation of antioxidant status and hematological parameters. Sections of the heart and aorta were examined for histological alterations. Results: Treatment with extract of Cyperus esculentus resulted in a significant increase (P<0.05) in superoxide dismutase, catalase and gluthathione peroxidase activities and a significant reduction in malondialdehyde level when compared to the untreated animals. However, the alterations observed in the hematological parameters were not improved on treatment with Cyperus esculentus. Conclusion: Treatment with medium (300mg/kg B.W) and high (600mg/kg B.W) of Cyperus esculentus reversed the histological alterations observed in the heart and aorta of the untreated animals. The study demonstrated that aqueous extract of Cyperus esculentus can protect against oxidative stress resulting from cadmium toxicity.

Keywords: Antioxidant status, cadmium, Cyperus esculentus, heart, hematology, histology


How to cite this article:
Innih SO, Eluehike N, Francis B. Effects of aqueous extract of Cyperus esculentus (tiger nut) on antioxidant status and hematological indices in the heart of cadmium-induced wistar rats. Niger J Exp Clin Biosci 2021;9:17-24

How to cite this URL:
Innih SO, Eluehike N, Francis B. Effects of aqueous extract of Cyperus esculentus (tiger nut) on antioxidant status and hematological indices in the heart of cadmium-induced wistar rats. Niger J Exp Clin Biosci [serial online] 2021 [cited 2021 Oct 24];9:17-24. Available from: https://www.njecbonline.org/text.asp?2021/9/1/17/316526




  Introduction Top


Cadmium (Cd) is a heavy metal that accumulates in the environment as a result of increased usage in the industries causing occupational and health hazards. Cd gets to the human body through foods, water, and undergo bioaccumulation threatening the well-being of human.[1],[2] Cd is accumulated in its soluble form as Cd chloride (CdCl2) and affects several organs including the heart, liver, and kidney. Cd affects the cardiovascular system by inducing hypertension[3] and diabetes,[4] causing direct toxic impact on gene transcription in the vascular endothelium.[5] It also increases vascular intima media thickness,[6] and myocardial infarction,[7] Cd accumulates in the wall of the aorta.[8] Direct myocardial structural damage of the heart has also been reported.[9]

Cd induced toxicity is mediated by the generation of free radicals and subsequent reactive oxygen species accumulation, and which in turn lead to decreased antioxidant level.[10] Oxidative stress, cell cycle progression, DNA damage, and apoptosis are the main events that prompts Cd poisoning.[11] The heart is a vital organ and is the most affected as it is more vulnerable to oxidative stress than most other tissues. Cd exerts cardiotoxicity at concentrations as low as 0.1 mM.[12]

Medicinal plants and their products have been used for the treatment of various diseases for centuries. There are several evidences available to show the beneficial effects of natural/herbal formulations and phytonutrients for its protection against heavy metal-mediated toxicity.[13],[14],[15],[16],[17]

Tiger-nut (Cyperus esculentus L.) is a tuber belonging to the family, Cyperaceae. It has a slightly sweet and nutty flavor.[18],[19] It is referred to as earth almond, zulu nut. In Nigeria, it is called Aya in Hausa, ofio in Yoruba and Akiausa in Igbo. Three varieties of this nut have been identified (black, brown, and yellow). The yellow variety is preferred as it gives more milk upon extraction. It also contains lower fat, less anti-nutritional factors, especially polyphenols and more protein.[20]

Tiger nut contains high amount of Vitamin B1, C, and E and minerals such as calcium, magnesium, and iron.[19],[21],[22],[23] Numerous studies have reported on the beneficial effect of this nut. The hyperglycemic,[24] anti-diabetic,[25] anticancer,[26],[27],[28] hepatoprotective[29] oil from the nut help prevents heart attack, thrombosis, and activates blood circulation,[30] treatment of urinary tract infection and colon cancer.[31]

No report exists that shows the role of C. esculentus in Cd-induced toxicity in the heart of rats; hence, this study was designed to investigate the protective effect of C. esculentus in Cd-induced alterations on antioxidant status and hematological indices in Wistar rats.


  Materials and Methods Top


Collection of plant materials and preparation of extracts

Dried Yellow tiger nuts (C. esculentus) were obtained from a local market in Benin City. The nuts were identified and authenticated at the Department of Plant Biology and Biotechnology University of Benin, Benin City. The bad nuts were selected out and removed after which the nuts were pulverized. Approximately 700 g of the powdered nuts were soaked in 3 L of distilled water for 48 h. The extract was filtered using Whitman filter paper No. 1, evaporated to dryness using a rotary evaporator, and then freeze dried using a freeze dryer. The extract was stored in an air-tight container and kept in the refrigerator at 4°C until use.

Experimental animals

A total of thirty male adult Wistar rats weighing 150–220 g were procured from the Animal House, Department of Anatomy, University of Benin. The rats were housed in conventional wire mesh cages under the standard laboratory conditions. They were allowed free access to water and pellet feed throughout the period of the experiment. Treatment of the animals was in accordance with the guidelines for animal care of the National Institute of Health (Institute of Laboratory Animal Resources Committee, 1985). The research ethics committee guideline principles and consent on the handling of animals of the College of Medicine, University of Benin (CMR/REC/2014/57), was obtained, adopted and strictly adhered to.

Experimental design

The thirty male Wistar rats were randomly divided into six groups of five animals each.

  • Group A: normal control animals (fed normal rat chow)
  • Group B: served as the negative control. Received CdCl2 daily (10 mg/kg body weight orally) for 60 days
  • Group C: received CdCl2 daily (10 mg/kg body weight orally) and treated with low dose (150 mg/kg body weight) of aqueous extract of tiger nuts for 60 days
  • Group D: received CdCl2 daily (10 mg/kg body weight orally) and treated with medium dose (300 mg/kg body weight) of aqueous extract of tiger nuts for 60 days
  • Group E: received CdCl2 (10 mg/kg body weight orally) and treated with high dose (600 mg/kg body weight) of aqueous extract of tiger nuts for 60 days
  • Group F: received CdCl2 daily (10 mg/kg body weight orally) and treated with standard drug (atorvastatin - 1.2 mg/kg body weight) for 60 days.


At the end of the experimental period, all animals were fasted overnight and sacrificed under deep anesthesia using chloroform. Blood was collected from the animals into plain sample tubes by cardiac puncture and separated to get serum by centrifuging at 3000 g for 10 min. Heart and aorta tissues were harvested and fixed immediately in 10% formal saline for histhopathological studies.

Evaluation of lipid peroxidation

The malondialdehyde (MDA) level was used to estimate the level of lipid peroxidation. MDA level was determined by the method of Varshney and Kale.[32]

Assay for antioxidant enzymes

Catalase (CAT) activity was determined in the serum by the method of Cohen et al.[33] Superoxide dismutase (SOD) activity was determined using the method of Misra and Fridovich.[34] Gluthathione peroxidase was assayed using the method of Addy and Goodman.[35]

Hematological parameters

Red blood cell (RBC) count, white blood cell (WBC) count, platelets count, packed cell volume, hemoglobin, and differential leukocyte count were assayed for using the automated hemoanalyzer.

Histhopathological examinations

Excised heart and aorta were analyzed for histological alterations after fixing for 2 days in 10% buffered formalin. Sections (5 μm thick) were paraffin embedded and stained with hematoxylin and eosin. The sections of the heart and aorta were obtained and examined using a microscope with a digital camera attached. Digital photomicrographs of the tissue sections were taken at × 100 magnifications.

Statistical analysis

The values obtained for antioxidant enzymes and hematological parameters were expressed as mean ± standard error of the mean. One-way analysis of variance, followed Tukey's comparison test using Graph pad prism 8.0 was performed for the comparison among the groups. Values of P < 0.05 were assumed statistically significant.


  Results Top


Effect of Cyperus esculentus on the body weight of cadmium-induced oxidative stress in rats

A nonsignificant change (P < 0.05) was observed in the body weight of the rats at day 60 when compared with their corresponding weight at day 0. The result is shown in [Figure 1] and [Figure 2].
Figure 1: Effect of Cyperus esculentus on the body weight of cadmium induced oxidative stress the rats. Values are mean ± standard error of the mean, n = 5 rats in each group

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Figure 2: Effect of Cyperus esculentus on the heart weight of cadmium induced oxidative stress the rats. Values are mean ± standard error of the mean, n = 5 rats in each group

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Effect of Cyperus esculentus on antioxidant status and lipid peroxidation of cadmium induced oxidative stress in rats

There was a significant decrease in the activities of SOD, CAT, and glutathione peroxidase (GPx) in the Cd-induced rats, when compared to the control group (P < 0.05). Treatment with the extract of C. esculentus resulted in a dose-dependent significant increase (P > 0.05) in SOD, CAT, and GPx levels when compared with the untreated rats. The results are shown in [Figure 3],[Figure 4],[Figure 5],[Figure 6].
Figure 3: Effects of Cyperus esculentus on superoxide dismutase in cadmium intoxicated rats. Values are mean ± standard error of the mean, n = 5 rats in each group

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Figure 4: Effects of Cyperus esculentus on catalase in cadmium intoxicated rats. Values are mean ± standard error of the mean, n = 5 rats in each group

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Figure 5: Effects of Cyperus esculentus on glutathione peroxidase cadmium intoxicated rats. Values are mean ± standard error of the mean, n = 5 rats in each group

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Figure 6: Effects of Cyperus esculentus on malondialdehyde in cadmium intoxicated rats. Values are mean ± standard error of the mean, n = 5 rats in each group

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Effect of Cyperus esculentus on hematological parameters of cadmium-induced oxidative stress in rats

The administration of Cd resulted in alteration in hematological indices: WBC count, RBC count, hematocrit percent, platelets, hemoglobin etc., administration of aqueous extract of C. esculentus did not normalize the alterations observed in these parameters.


  Discussion Top


In the environment, Cd is dangerous because humans consume both plants and animals that absorb Cd efficiently and concentrate it within their tissues. The precise mechanism for Cd-induced cardiac toxicity is not fully understood. It has been suggested that Cd may decrease or increase the level of nitric oxide in endothelial cells and may increase the production of reactive oxygen species resulting in lipid peroxidation[36],[37],[38] and this can lead to alteration of metabolic pathways involved in energy metabolism, protein synthesis, and antioxidant network.

The effect of Cd toxicity can be assessed by the measurement of cytoplasmic enzymes released into the blood stream during oxidative membrane damage. In this study, we assessed the levels of antioxidant enzymes, lipid peroxidation products, and hematological parameters to determine the effects of C. esculentus in Cd-induced rats.

Clinical diagnosis of Cd–induced cardiac toxicity entails the measurement of cardiac enzymes such as aspartate amino transferase level. We have established a significant increase in the aspartate amino transferase level in Cd induced rats compared to normal control (manuscript under review).

Cellular enzymatic antioxidants such as SOD, CAT, GPx are the first line of defense suppressing the formation of free radicals inside the cells during Cd-induced toxicity.[39] Several reports are available to show that Cd induces oxidative stress.[40],[41],[42] Cd contributes to the adverse effects of organisms probably because of its ability to induce oxidative stress through alterations in the activities of antioxidant enzymes CAT and SOD. The reduced level of SOD, observed in the negative control (Cd only) may be due to the fact that SOD being metalloenzyme, its Zn2+ ions has been replaced by Cd.

The significant reduction in the levels of SOD and CAT and GPx in the Cd administered group may be attributed to a devastating oxidative alteration of enzymatic proteins and bio-membrane lipids by reactive oxygen species. The significant reduction in CAT activity might be due to the facts that at physiological pH, the nitrogen atom of imidazole ring in His-74 is deprotonated and thus interacts with the Cd2+ ion hence leading to the reduction in CAT activity.[43] Treatment with aqueous extract of C. esculentus resulted in a significant elevation in the activities of these antioxidant enzymes [Figure 3]. Oil extracted from the tiger nut have been shown to contain various radical-scavenging antioxidants.[44] Tiger nuts contains high amount of Vitamin E and C which are important antioxidants hence may have been responsible for the observed antioxidant effects recorded in this study. The significantly increased antioxidants recorded in this study might also be due to the presence of water-soluble flavonoid glycosides found in high amount in tiger nut.[45] The lipid peroxidation level is an important parameter in the measurement of toxicity of xenobiotics like Cd as it serves as one of the most important indicator of oxidative stress and damage. The increased level of MDA, a reliable marker of lipid peroxidation observed in this study is in line with the results from other studies [Figure 3]. Cd-induced lipid peroxidation has been reported in previous study.[46] The significant reduction in MDA level observed in C. esculentus treated rats when compared to the untreated group support the claim that natural antioxidant like tiger nut can effectively suppress lipid peroxidation.[47]

Research has shown that hematopoeisis has been reported to be adversely affected in Cd induced conditions,[48] leading to severe anemia in addition to with marked suppression of erythropoietin production. Similarly, the immune system suffers from Cd-induced impairment at several levels. Lymphocyte proliferation and natural killer cell activity are also suppressed by Cd. This report confirms the alterations observed in the hematological indices in this study. Although the administration of extracts of C. esculentus did not normalize the alterations in WBC, RBC, Hb, HCT, and other hematological indices assayed for in this study [Table 1].
Table 1: Effects of Cyperus esculentus on some hematological parameters in cadmium intoxicated rats.

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Histological evaluation of the heart tissue revealed that Cd treatment caused severe vascular stenosis and perivascular infiltrates of inflammation as seen in the untreated animals. Treatment of the animals with low dose (150 mg/kg body weight of C. esculentus) resulted in a characteristic mild vascular ulceration and perivascular infiltrates in the cardiac architecture. On the other hand, medium (300 mg/kg body weight) and high dose (600 mg/kg body weight) treatment with the extract of C. esculentus was capable of preventing the damage caused by Cd on the myocardium as evident by the normal architecture observed in this group. Priya et al.,[49] and Oyinloye et al.[50] demonstrated in their study that treatment with extracts from Tinospora cordifolia and Sesamum indicum seeds, respectively, prevented the histological damage caused by Cd on the heart[Plates 1],[Plates 2],[Plates 3],[Plates 4],[Plates 5],[Plates 6],.



Histhopathology of the aorta also revealed that medium and high dose treatment with C. esculentus prevented the severe ulceration and asymmetric hypertrophy seen in the untreated animals[Plates 7],[Plates 8],[Plates 9],[Plates 10],[Plates 11],[Plates 12].




  Conclusion Top


This study has established that treatment of Cd-induced rats with medium and high doses of aqueous extract of C. esculentus (tiger nut) can ameliorate oxidative stress and help prevent damage to the heart and aorta induced by Cd toxicity.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
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