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 Table of Contents  
ORIGINAL ARTICLE
Year : 2018  |  Volume : 2  |  Issue : 4  |  Page : 276-280

Scavenging antioxidative bioactivities of cucumber (Cucumis sativus) fruit juice in rabbits overdosed with amoxicillin


1 Department of Medical Laboratory Science, Edo University, Iyamho, Nigeria
2 Department of Medical Laboratory Science, Achievers University, Owo, Nigeria

Date of Submission09-Sep-2018
Date of Decision18-Sep-2018
Date of Acceptance15-Oct-2018
Date of Web Publication11-Dec-2018

Correspondence Address:
Prof. Mathew Folaranmi Olaniyan
Department of Medical Laboratory Science, Edo University, Iyamho
Nigeria
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/bbrj.bbrj_119_18

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  Abstract 


Background: Raw cucumber (Cucumis sativus) fruit juice contains phytochemicals and phytonutrients of antioxidative bioactivities while overdose of amoxicillin could bring about drug-induced toxicity and oxidative stress. Malondialdehyde (MDA) is an oxidative index while catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) are scavenging antioxidants. This work was therefore designed to determine the scavenging antioxidative bioactivities of cucumber (C. sativus) fruit juice in rabbits overdosed with amoxicillin. Methods: Fifteen rabbits of the same sex divided into three groups of 5 rabbits each were used for the study. Group A – 5 control rabbits; Group B – 5 rabbits given 30 mg/kgBW subcutaneous injection of amoxicillin every 24 h for 7 days which was followed by 30 ml raw cucumber fruit juice supplementation for 14 days; and Group C – 5 rabbits given 30 mg/kgBW subcutaneous injection of amoxicillin every 24 h and raw cucumber fruit juice supplementation for 14 days simultaneously. Plasma MDA, CAT, SOD, and GPx were determined in the rabbits biochemically by spectrophotometry, fluorometry, and ELISA. Results: The results obtained showed a significant increase in plasma value of MDA and a significant decrease in plasma CAT, SOD, and GPx when the rabbits were given 30 mg/kgBW subcutaneous injection of amoxicillin (P < 0.05). These significant biochemical changes were however reversed in the rabbits when they were supplemented with 30 ml raw cucumber fruit juice (P < 0.05). Simultaneous administration of 30 mg/kgBW subcutaneous injection of amoxicillin and 30 ml raw cucumber fruit juice showed no significant biochemical changes (P > 0.05). Conclusion: This work showed abnormal plasma values of MDA, CAT, SOD, and GPx following amoxicillin overdose which was reversed owing to possible scavenging antioxidative potentials of raw cucumber fruit juice. The raw cucumber juice could therefore be a natural antidote in drug-induced toxicity.

Keywords: Amoxicillin overdose, catalase, cucumber (Cucumis sativus) fruit juice, glutathione peroxidase, malondialdehyde, scavenging, superoxide dismutase


How to cite this article:
Olaniyan MF, Afolabi T. Scavenging antioxidative bioactivities of cucumber (Cucumis sativus) fruit juice in rabbits overdosed with amoxicillin. Biomed Biotechnol Res J 2018;2:276-80

How to cite this URL:
Olaniyan MF, Afolabi T. Scavenging antioxidative bioactivities of cucumber (Cucumis sativus) fruit juice in rabbits overdosed with amoxicillin. Biomed Biotechnol Res J [serial online] 2018 [cited 2019 Jan 16];2:276-80. Available from: http://www.bmbtrj.org/text.asp?2018/2/4/276/247241




  Introduction Top


Cucumber (Cucumis sativus) is extremely beneficial for overall health; it could relieve thirst as it is mostly made of water and important nutrients that are essential for the human body. The flesh of cucumber is rich in Vitamin A, Vitamin C, K, and folic acid, while the hard skin is rich in fiber and a range of minerals including magnesium, molybdenum, and potassium[1],[2] In addition, cucumber contains silica, a trace mineral that contributes greatly to strengthening our connective tissues.[3],[4],[5] There are traditional but nonscientific applications of cucumber to heal many skin problems, manage diabetes, control blood pressure, prevent constipation and kidney stones, and sunburn. Cucumber also contains ascorbic and caffeic acids which prevent water loss, therefore, it is frequently applied topically to burns and dermatitis. Cucumber has excellent cleaning properties, and they actively remove accumulated waste and toxins from the body. Drinking cucumber juice can result in the development of strong and healthy bones and teeth due to its Vitamin K content. Cucumber is a rich source of cucurbitacins which blocks cancer cell development according to scientists.[3],[4],[5]

Cucumber has high alkaline levels, thus regulating the body blood pH and neutralizing acidity. People with gastric issues should consume cucumbers frequently. Since cucumbers contain mostly water, they are excellent food to help cleanse and wash away toxins.[6],[7] Toxins are often stored in fat cells and cause inflammation. But beyond just water content, cucumbers also supply electrolytes to restore hydration of the body cells and balance the fluid levels in the body. This makes cucumber an important food to aid in weight loss and other inflammatory conditions such as arthritis.

Cucumber juice is a diuretic, so it is able to prevent kidney stones too. It can also counter the effects of uric acid, which prevents inflammation from conditions such as arthritis, asthma, and gout[8],[9] The phytochemical constituents of cucumber fruit include flavonoids, tannin, saponins, alkaloid, and steroids.[1],[2]

Amoxicillin is an antibiotic used to treat bacterial infection such as acute otitis media, streptococcal pharyngitis, pneumonia, skin infections, urinary tract infections,  Salmonella More Details infections, Lyme disease, and Chlamydia infections.[10],[11],[12] It is effective in the treatment of stomach infections of Helicobacter pylori. It is a moderate-spectrum, bacteriolytic, β-lactam antibiotic in the aminopenicillin family used to treat susceptible Gram-positive and Gram-negative bacteria.[10],[11],[12] Side effects include nausea, vomiting, rashes, antibiotic-associated colitis possibly abnormal liver function, anemia, pancytopenia, drug-induced hepatitis, and oliguria.[13]

This work was therefore designed to determine the scavenging antioxidative bioactivities of cucumber (C. sativus) fruit juice in rabbits overdosed with amoxicillin.


  Methods Top


Study area

This work was carried out at the Animal house of Achievers University is in Owo, Ondo State, Nigeria. The university is a private-sector initiative, established in 2007 and accredited by the National Universities Commission. It is located on land in the Idasen community of Owo, consisting of Ulale 1, Ulale 11, Ulema, Ijegunma, Isijogun, and Amurin Elegba (formerly Amurin, Ogain). The university sprang from the Achievers Group of Education and Training Organization, located in Ibadan Oyo State of Nigeria owned and run by Hon. Dr. Bode Ayorinde and other educationalists. The university commenced academic activities during the 2007/2008 academic session. In the Nigerian National University Commission annual university rankings for 2013, it was rated 53rd. It has three colleges which include College of Natural and Applied Sciences, College of Engineering and Technology, and College of Social Science, and Management including a Postgraduate school. The proposal was reviewed and approved by Research and an Ethical committee of the Department of Biological Sciences, Achievers University, Owo, Nigeria before the commencement of the work. (AUOBS/2018/31/91; 19th February 2018).

Study population

Fifteen rabbits of the same sex divided into three groups of 5 rabbits each were used for the study. The rabbits were bought animal farm in Owo, Nigeria and was presented to Federal School of Agriculture, Akure, for confirmation.

  • Group A – 5 control rabbits were fed with normal meal and water throughout the period of investigation
  • Group B – 5 rabbits given 30 mg/kgBW subcutaneous injection of amoxicillin every 24 h for 7 days which was followed by 30 ml raw cucumber fruit juice supplementation for 14 days
  • Group C – 5 rabbits given 30 mg/kgBW subcutaneous injection of amoxicillin every 24 h and raw cucumber fruit juice supplementation for 14 days simultaneously.


Administration of amoxicillin

Amoxicillin was bought from a registered pharmaceutical shop in Owo, Nigeria. An overdose of 30 mg/kgBW of amoxicillin was given through subcutaneous injection every 24 h for 7 days.

Preparation of cucumber (Cucumis sativus) fruit juice

Cucumber (C. sativus) was bought from fruit vendors in Owo, Nigeria. The fruit was presented to Federal School of Agriculture, Akure, for confirmation. The fruit was washed in sterile water and then sliced. The sliced fruit was blend together using an electronic blender. The raw fluid was extracted using a sterile sieve. The raw liquid extract was served to the rabbits as juice. 30 ml was given to the rabbits on daily basis. The juice will always be freshly prepared.

Specimen (blood) collection

Five milliliters of venous blood was collected from each of the rabbits into lithium heparinized bottles for the biochemical analysis.

Biochemical analysis

Estimation of plasma oxidative biomarkers

Lipid Peroxidation (Malondialdehyde)

This is determined by measuring the thiobarbituric acid reactive substance produced during lipid peroxidation, using the reagent kit of Abcam kit.

Principle

This method is based on the reaction between 2-thiobabituric acid and malondialdehyde (MDA), and product of lipid peroxidation. On heating in an acidic pH, the product is a pink color complex which is absorb maximally at 523 nm and which is extractable into organic solvents such as butanol. The results are expressed as a result of free MDA produced.

Catalase using Randox kit

CAT activity was measured quantitatively by the method of von Euler and Josephson by allowing the enzyme solution to react with hydrogen peroxide for varying periods of time and measuring the excess peroxide remaining by titration with potassium permanganate.

Principle

Catalase enzyme is present in nearly all animal cells, plant and bacteria. It acts to prevent accumulation of noxious H2O2 by influencing the conversion of H2O2 to O2 and H2O.

i.e., H2O2 + H2O2 2H2O + O2

Peroxidase, which is less widely distributed catalyze the above reaction. The absorbance is read at 480 nm. At high concentration of low molecular weight alcohols or formaldehyde and low peroxide concentration, CAT exhibits peroxidative activity. Absorbance is read at 480 nm 30–60 s against distilled water.

Superoxide dismutase activity

This was determined using Abcam kit.

Principle

Adrenaline auto-oxidizes rapidly in aqueous solution to adrenochrome, whose concentration can be determined at 420 nm using a spectrophotometer. The auto-oxidation of adrenaline depends on the presence of superoxide anions. The enzyme SOD inhabits the auto-oxidation of adrenaline by catalyzing the breakdown of superoxide anions. The degree of inhibition is thus a reflection of the activity SOD and is determined at one unit of the enzyme activity. It was mixed and absorbance was read at 420 nm.

Glutathione peroxidase

This was determined using Abcam kit.

Principle

Glutathione peroxidase (GPX) catalyzes the reduction of an organic peroxide by oxidizing reduced glutathione (GSH) to form disulfide glutathione (GSSG). The oxidized glutathione is then reduced by glutathione reductase (GR) and b-nicotinamide adenine dinucleotide phosphate (NADPH) forming NADP + (resulting in decreased absorbance at 340 nm) and recycling the GSH. Since glutathione peroxide (GPX) is limiting, the decrease in absorbance at 340 nm is directly proportional to the glutathione (GPX) concentration.

Method of data analysis

A Statistical Package for Social Sciences 18.0 version was used for the analysis of the data appropriately. The level of significance was taken at 95% confidence interval and P < 0.05 was considered statistically significant.


  Results Top


The results obtained showed a significant increase in the plasma value of MDA in the rabbits given overdose of 30 mg/kgBW subcutaneous injection of amoxicillin every 24 h for 7 days. Compared with the results obtained from the control rabbits, their basal samples and when the same rabbits were thereafter supplemented with 30 ml of raw cucumber fruit juice for 14 days with P < 0.05 [Table 1], [Table 2] and [Figure 1], [Figure 2].
Table 1: Mean and standard deviation of plasma values of malondialdehyde, superoxide dismutase, and glutathione peroxidase obtained in the rabbits

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Table 2: Comparative analysis of the mean and standard deviation of plasma values of malondialdehyde, superoxide dismutase, and glutathione peroxidase obtained in the rabbits

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Figure 1: Comparative description of the mean and standard deviation of plasma values of superoxide dismutase and glutathione peroxidase obtained in the rabbits

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Figure 2: Comparative description of the mean and standard deviation of plasma values of malondialdehyde obtained in the rabbits

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The results obtained also showed a significant decrease in the plasma value of CAT, SOD, and GPx in the rabbits given overdose of 30 mg/kgBW subcutaneous injection of amoxicillin every 24 h for 7 days compared with the results obtained from the control rabbits, their basal samples and when the same rabbits were thereafter supplemented with 30 ml of raw cucumber fruit juice for 14 days with P < 0.05 [Table 1], [Table 2] and [Figure 1], [Figure 2].

There was a significant decrease in the plasma value of MDA and a significant increase in the plasma value of CAT, SOD, and GPx in the rabbits supplemented with 30 ml of raw cucumber fruit juice for 14 days after amoxicillin overdose with P < 0.05 [Table 1], [Table 2] and [Figure 1], [Figure 2].

There was no significant difference in the plasma value of MDA, CAT, SOD, and GPx in the results obtained from the basal samples, when the rabbits were supplemented with 30 ml of raw cucumber fruit juice and the control with P > 0.05 [Table 1], [Table 2] and [Figure 1], [Figure 2].

There was no significant difference in the plasma value of MDA, CAT, SOD, and GPx in the results obtained in the rabbits following coadministration of overdose of 30 mg/kgBW subcutaneous injection of amoxicillin every 24 h for 7 days and supplement of 30 ml of raw cucumber fruit juice for 14 days compared with the results obtained from the control rabbits and their basal samples P > 0.05 [Table 1], [Table 2] and [Figure 1], [Figure 2].


  Discussion Top


The results obtained showed a significant increase in the plasma value of MDA in the rabbits given overdose of 30 mg/kgBW subcutaneous injection of amoxicillin every 24 h for 7 days compared with the results obtained from the control rabbits, their basal samples and when the same rabbits were thereafter supplemented with 30 ml of raw cucumber fruit juice for 14 days. This could be explained as the normal dose of amoxicillin is 7.5 mg/kgBW subcutaneous injection[14] while 30 mg/kgBW subcutaneous injection of amoxicillin is an overdose which must have subjected the affected rabbits to oxidative stress as indicated by the increase in plasma level of MDA. This is because MDA occurs naturally and is a marker for oxidative stress.[15],[16]

The results obtained also showed a significant decrease in the plasma value of CAT, SOD, and GPx in the rabbits given overdose of 30 mg/kgBW subcutaneous injection of amoxicillin every 24 h for 7 days compared with the results obtained from the control rabbits, their basal samples and when the same rabbits were thereafter supplemented with 30 ml of raw cucumber fruit juice for 14 days. CAT, SOD, and GPx are scavenging antioxidant whose level decreases in oxidative stress. CAT can also catalyze the oxidation, by hydrogen peroxide, of various metabolites and toxins, including formaldehyde, formic acid, phenols, acetaldehyde, and alcohols. It does so according to the following reaction: H2O2 + H2R → 2H2O + R.[17],[18],[19],[20],[21] Superoxide is one of the main reactive oxygen species in the cell. As a consequence, SOD serves a key antioxidant role.[22],[23],[24] GPx is the general name of an enzyme family with peroxidase activity whose main biological role is to protect the organism from oxidative damage. The biochemical function of GPx is to reduce lipid hydroperoxides to their corresponding alcohols and to reduce free hydrogen peroxide to water.[25],[26],[27],[28],[29]

There was a significant decrease in the plasma value of MDA and a significant increase in the plasma value of CAT, SOD, and GPx in the rabbits supplemented with 30 ml of raw cucumber fruit juice for 14 days after amoxicillin overdose. This could be attributed to the fact that Cucumber fruit juice contains a wide variety of biologically active, nonnutritive compounds known as phytochemicals such as alkaloids, flavonoids, tannins, phlobatannins, steroids, saponins, and phytonutrients such as Vitamin C.[30] These phytochemicals and phytonutrients have health benefits bioactivities beyond basic nutrition[31] with the antioxidants having potential to reduce toxins and the risk of several deadly diseases in man.[32] Fisetin a constituent of cucumbers has a significant role in brain health, protecting the brain's nerve cells, and memory from declining and preventing learning impairments associated with Alzheimer's.[30]

In general, the presence of flavonoids and tannins in the extract as evidenced by preliminary phytochemical screening suggests that these compounds might be responsible for free radical scavenging and analgesic effects.[1]


  Conclusion Top


This work showed a significant increase in plasma value of MDA and a significant decrease in the plasma value of CAT, SOD, and GPx following amoxicillin overdose. This abnormal biochemical alteration was reversed owing to possible scavenging antioxidative potentials of raw cucumber fruit juice. The raw cucumber juice could, therefore, be a natural antidote in drug-induced toxicity.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
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    Tables

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