Full Project – Effect of garlic and ginger co-administration on serum glutathione levels of Wistar rat exposed to vanadium, lead and arsenic

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ABSTRACT

Oxidative stress in blood and other soft tissues has been postulated to be one of the possible mechanisms of heavy metal-induced toxic effects. Therefore, this present study aims at evaluating the abrogative effects of garlic and ginger co-administration on serum antioxidants in wistar rats exposed to arsenic (As), vanadium (V) and lead (Pb) by assaying for the serum glutathione (GSH) level. Thirty-five wistar rats weighing 100-150g were randomly divided into seven (7) groups of five (5) animals each and were administered treatments as follows; Group A (normal control) received distilled water only; group B (HM only) received sodium arsenite (2.5 mg/kg b.wt), lead acetate (60 mg/kg b.wt) and vanadium pentaoxide (9.8 mg/kg b.wt); group C (HM + GI1 + GA1) received HM + ginger (100 mg/kg b.wt) + garlic (100 mg/kg b.wt); group D (HM + GI2 + GA2) recieved HM + ginger (200 mg/kg b.wt) + garlic (200 mg/kg b.wt); group E (HM + VITC) recieved HM + vitamin C (100 mg/kg b.wt); group F (GI1 + GA1) received ginger (100 mg/kg b.wt) + garlic (100 mg/kg b.wt) and group G (GI2 + GA2) recieved ginger (200 mg/kg b.wt) + garlic (200 mg/kg b.wt). All treatments were orally administered daily, for four weeks. From the results of this study, arsenic, lead and vanadium exposure provoked a 26.3% increase the GSH level when compared to the normal control group. Treatment of heavy metal-induced toxicity with low (100 mg/kg b.wt) and high (200 mg/kg b.wt) doses of ginger and garlic cocktail was able to elicit a 55.6% significant (p < 0.05) increase, and a 44.2% increase respectively, in the serum GSH levels compared to the HM only group. Vitamin C, which is a potent antioxidant exhibited similar properties, as it significantly (p < 0.05) increased the serum GSH levels by 62.5%. From the study ginger (Zingiber offcinale) and garlic (Allium sativum) were also shown to enhance the GSH levels of a normal rat and its efficiency almost on par with vitamin C, a well-known potent antioxidant.

Keywords: Oxidative stress, Ginger, Garlic, Heavy metals, Glutathione and wistar rat

CHAPTER ONE

INTRODUCTION

1.1 Background of the Study

Heavy metals are defined as metallic elements that have a relatively high density compared to water. With the assumption that heaviness and toxicity are inter-related, heavy metals also include metalloids, such as arsenic, that are able to induce toxicity at low level of exposure (De Larco et al., 2004). In recent years, there has been an increasing ecological and global public health concern associated with environmental contamination by these metals. Also, human exposure has risen dramatically as a result of an exponential increase of their use in several industrial, agricultural, domestic and technological applications (Eric Block, 2010). The sources of heavy metals in the environment include geogenic, industrial, agricultural, pharmaceutical, domestic effluents, and atmospheric sources. Environmental pollution is very prominent in point source areas such as mining, foundries and smelters, and other metal-based industrial operations (Melino et al., 2010).

Although heavy metals are naturally occurring elements that are found throughout the earth’s crust, most environmental contamination and human exposure result from anthropogenic activities such as mining and smelting operations, industrial production and use, and domestic and agricultural use of metals and metal-containing compounds (ATSDR, 2003). Environmental contamination can also occur through metal corrosion, atmospheric deposition, soil erosion of metal ions and leaching of heavy metals, sediment re-suspension and metal evaporation from water resources to soil and ground water (ATSDR, 2003). Natural phenomena such as weathering and volcanic eruptions have also been reported to significantly contribute to heavy metal pollution (De Larco et al., 2004). Industrial sources include metal processing in refineries, coal burning in power plants, petroleum combustion, nuclear power stations and high tension lines, plastics, textiles, microelectronics, wood preservation and paper processing plants.

The essential heavy metals exert biochemical and physiological functions in plants and animals. They are important constituents of several key enzymes and play important roles in various oxidation-reduction reactions (Wilson et al., 2007). Copper for example serves as an essential co-factor for several oxidative stress-related enzymes including catalase, superoxide dismutase, peroxidase, cytochrome c oxidases, ferroxidases, monoamine oxidase, and dopamine β-monooxygenase (Wilson et al., 2007). While some heavy metal such as vanadium, lead and arsenic are toxic for human health. Heavy metal-induced toxicity and carcinogenicity involves many mechanistic aspects, some of which are not clearly elucidated or understood (Ramos et al., 2006). However, each metal is known to have unique features and physic-chemical properties that confer to its specific toxicological mechanisms of action (Petrusevski et al., 2007).

Garlic and aged garlic extract exhibit direct antioxidant effects and to enhance the serum levels of two antioxidant enzymes: catalase and glutathione peroxidase (Melino et al., 2010). Garlic (both the homogenate of 10% in physiological saline solution and its supernatant) was able to reduce the radicals present in cigarette smoke. It was also hypothesized that garlic organo-sulfur compounds may be able to prevent depletion of glutathione. Garlic ingestion may protect patients who experience increase in reactive oxygen induced stress on liver function (De Larco et al., 2004). Given the extreme chronic stress many people now face in their daily life, garlic may prove usefulness in countering the negative impact of this stress on human physiology. Antioxidants of garlic help protect cells from damage caused by free radicals. Free radicals are molecules produced when your body breaks down food or when you’re exposed to harmful environmental factors, so ingesting foods high in antioxidants may help combat those negative effects on the body (Wilson et al., 2007).

Ginger is known to exhibit a powerful antioxidant activity due to its oil which has protective effect on DNA. This effect has been demonstrated in some cell culture. Ginger has preventive effect on lipid peroxidation and also inhibits or breaks its chain (ATSDR, 2003). Ginger modulates genetic pathway, acts on tumor suppression of genes and modulates some biological activities. Gingerols and Paradol have been reported to have good anti-platelet and COX-I inhibitory properties. Ginger acts and protect gastric mucosa against several ulcerogenic agents and is very useful in cases of ulcerogenesis because of its antioxidant properties (Melino et al., 2010). Ginger shows strong antiemetic property by enhancing intestinal motility and inhibiting serotonin receptors. Ginger was reported to stimulate the peripheral anti-cholinergic and anti-histaminic receptors and antagonize 5-hydroxytreptamine receptors in the GIT. Ginger has long been revered as a natural remedy for upset stomachs, especially by pregnant women suffering from morning sickness. Ginger candies, natural ginger ale, and ginger tea are easy ways to get ginger into your system when you’re feeling a little green (Eric Block, 2010).

1.2 Aim and Objective

1.2.1 Aim

The aim of this project work is to determine the effects of garlic and ginger co-administration on serum antioxidants of wistar rats exposed to arsenic, vanadium and lead.

1.2.2 Objective

The objectives of this study are as follow:

To prepare ginger and garlic extract
To induced toxicity by exposing the wistar rat to arsenic, vanadium and lead
To isolate the serum (supernatant of the blood sample)
To evaluate the levels of glutathione (GSH) in the serum of the experimental animal

1.3 Problem Statement

Heavy metal poisoning from naturally occurring heavy metal compounds in drinking water remains a problem in many parts of the world (El-Aasr et al., 2010).Heavy metal exposure can result in both acute and chronic toxicity in humans. Acute heavy metal poisoning is relatively less common but has been documented after accidental ingestion of insecticides or pesticides, and attempted suicides or murders with arsenicals (El-Aasr et al., 2010). Heavy metal poisoning is a medical condition caused by increased levels of the element arsenic in the body. It is a chronic illness resulting from drinking water with high levels of heavy metal over a long period of time (such as from five to twenty years) (Melino et al., 2010). In spite of the awareness that alliums (including ginger and garlic) are rich in antioxidant properties such as ginger and garlic respectively, there is however, inadequate information about the ameliorating effects of these alliums on arsenic toxicity when used as dietary supplement (Wilson et al., 2007).

Few studies have been done on dietary intervention for heavy metal poisoning. This may be due to the cumbersomeness and wide range in the assessment of the chemical components in the food. Heavy metals are uses in the production of pesticides (treated wood products), herbicides and insecticides (Jimoh et al., 2008). Garlic and ginger are extremely helpful to the body as they are natural detoxifiers. With advances in technology and the recent development of animal models for heavy metal carcinogenicity, understanding of the toxicology of heavy metal will continue to improve (Jimoh et al., 2008).

1.4 Justification/ Rationale of the Study

Several studies have been conducted on heavy metal exposure and poisoning from different sources but few studies have been carried out on the detoxification of heavy metal poisoning using dietary intervention and most importantly using the plant such as ginger and

garlic to ameliorate heavy metal toxicity (Petrusevski et al., 2007). All age groups of human beings are vulnerable to heavy metal poisoning due to its natural occurrence in groundwater. There is a need to establish a dietary intervention strategy in detoxifying heavy metal poisoning. This justifies the use of crops in the die since they are mostly eaten by humans as spices in food. Therefore, this research topic is worthy of study and may make a significant contribution to the body of already existing knowledge on food toxicology (Petrusevski et al., 2007).

1.5 Public Health Significance of the Study

Heavy metals are natural contaminant of ground water, as well as drinking water, which translates into a public health issue worldwide. As the world population increases, one of the most fundamental resources for human survival, clean water, is decreasing (Schmeisser et al., 2006). This study is of public health significance because all age groups of human beings are vulnerable to heavy metal poisoning due to its natural occurrence in groundwater and various anthropogenic sources. The heavy metal is a natural environmental contaminant to which humans are routinely exposed in food, water, air, and soil. There is therefore a need to establish a dietary intervention strategy in detoxifying heavy metal poisoning. Ginger and garlic both have antioxidant and anticancer properties among other properties they possess. Their dietary intervention in detoxifying heavy metal poisoning is worthy of research (Schmeisser et al., 2006).

1.6 Limitation

The inability to estimate the various bioactive components present in Garlic and ginger

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