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ORIGINAL ARTICLE
Year : 2019  |  Volume : 6  |  Issue : 1  |  Page : 17-21

Assessment of plasma iron, transferrin alanine, and aspartate transaminase in amoxicillin overdose supplemented with raw cucumber juice


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

Date of Submission25-Aug-2018
Date of Acceptance15-Oct-2018
Date of Web Publication30-Apr-2019

Correspondence Address:
Dr. 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/jhrr.jhrr_41_18

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  Abstract 


Background: Raw cucumber (Cucumis sativus) fruit juice contains substances of health-promoting bioactivities. Elevated plasma alanine transaminase (ALT/serum glutamate-pyruvate transaminase), aspartate transaminase (AST/serum glutamic oxaloacetic transaminase), iron, and decreased transferrin could indicate hepatotoxicity, hepatitis, and kidney and liver damage, which could be drug induced. Aim and Objective: This work was designed to determine the benefits of cucumber fruit juice as a natural antidote in amoxicillin-induced hepatotoxicity by assessing the plasma iron, transferrin alanine, and AST in amoxicillin overdose supplemented with raw cucumber juice. Materials and Methods: Fifteen rabbits of the same sex weighing 0.9–1.4 kg divided into three groups of five rabbits each were used for the study. Group A – five control rabbits; Group B – five rabbits given 30 mg/kg body weight (BW) 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 – five rabbits given 30 mg/kg BW subcutaneous injection of amoxicillin every 24 h and raw cucumber fruit juice supplementation for 14 days simultaneously. Plasma ALT, AST, iron, and transferrin were determined in the rabbits by spectrophotometry and enzyme-linked immunosorbent assay. Results: The results obtained showed a significant increase in plasma ALT, AST, and iron and a significant decrease in plasma transferrin when the rabbits in Group B were given 30 mg/kg BW subcutaneous injection of amoxicillin (B2 samples) compared with the results obtained from their basal samples (B1) and control rabbits (A) and when they were given 30 ml of raw cucumber fruit juice supplementation (B3 samples) (P < 0.05). The results obtained showed a significant decrease in plasma ALT, AST, and iron and a significant increase in plasma transferrin when the Group B rabbits were given 30 ml of raw cucumber fruit juice (B3 sample) than when they were given subcutaneous injection of amoxicillin (B2 samples) (P < 0.05). Conclusion: This work revealed significant biochemical alterations in the values of iron, transferrin, AST, and ALT when given 30 mg/kg BW subcutaneous injection of amoxicillin and supplementation of raw cucumber fruit juice. Cucumber juice is therefore beneficial as a possible antidote to amoxicillin-induced hepatotoxicity.

Keywords: Amoxicillin, aspartate transaminase, cucumber juice, hepatotoxicity, plasma iron, transferrin alanine transaminase


How to cite this article:
Olaniyan MF, Adepoju DB. Assessment of plasma iron, transferrin alanine, and aspartate transaminase in amoxicillin overdose supplemented with raw cucumber juice. J Health Res Rev 2019;6:17-21

How to cite this URL:
Olaniyan MF, Adepoju DB. Assessment of plasma iron, transferrin alanine, and aspartate transaminase in amoxicillin overdose supplemented with raw cucumber juice. J Health Res Rev [serial online] 2019 [cited 2019 May 20];6:17-21. Available from: http://www.jhrr.org/text.asp?2019/6/1/17/257480




  Introduction Top


Cucumber (Cucumis sativus) juice is produced by squeezing a cucumber pod or pressing it on a sieve. Cucumber contains 98% of water.[1] Cucumber juice has large amounts of Vitamin A, Vitamin C, Vitamin K, calcium, potassium, silicon, copper, sterol, and soluble and insoluble fibers.[2] It is used in traditional medicine in the treatment of respiratory tract inflammation and to reduce lingering cough, soothe heartburn, and reduce acid in the stomach and burns including rashes.[3],[4] Cucumber juice is an insect repellant against wood lice and fish moths.[5],[6],[7]

Amoxicillin is an antibiotic used in the treatment of a number of bacterial infections. It is the first-line treatment for middle-ear infections. It may also be used for throat infection, pneumonia, skin infections, and urinary tract infection. It is taken by mouth, or less commonly by injection.[8] Side effects of the antibiotic include nausea; vomiting; rashes; antibiotic-associated colitis; loose bowel movements (diarrhea); mental changes; lightheadedness; insomnia; confusion; anxiety; sensitivity to lights and sounds; and unclear thinking; and acute overdoses of amoxicillin may cause lethargy, bleeding, vomiting, and renal dysfunction.[9],[10]

Aspartate transaminase (AST) or aspartate aminotransferase or serum glutamic oxaloacetic transaminase is a liver enzyme[11],[12] that catalyzes the reversible transfer of α-amino group[13],[14] between aspartate and glutamate and as such, is an important enzyme in amino acid metabolism.[15],[16],[17] This enzyme is found in the liver, heart, skeletal muscle, kidneys, brain, and red blood cells. Plasma aspartate and alanine aminotransferase levels including their ratio (AST/alanine transaminase [ALT] ratio) are common biomarkers of liver damage/disease.[18]

ALT is formerly referred to as serum glutamate-pyruvate transaminase. ALT is found in plasma and body tissues, but commonly in the liver.[19] ALT is commonly measured to determine hepatocellular injury.[20]

Plasma iron is the amount of circulating iron bound to transferrin. Transferrins are iron-binding blood plasma glycoproteins that control the level of free iron (Fe) in biological fluids. Nearly 65% of the iron in the body is bound up in hemoglobin; about 4% is in myoglobin; about 30% of the iron in the body is stored as ferritin or hemosiderin in the spleen, the bone marrow, and the liver; and small amounts of iron are found in other molecules in cells of the body.[21] Transferrin is produced by the liver and binds one or two iron (III) ions, i.e., ferric iron (Fe3+).[22]

This work was designed to determine the benefits of cucumber fruit juice as a natural antidote in amoxicillin-induced hepatotoxicity by assessing the plasma iron, transferrin alanine, and AST in amoxicillin overdose supplemented with raw cucumber juice.


  Materials and Methods Top


This prospective case–control study was carried out for 2 months at the Animal House of Achievers University, Owo, Nigeria. This study was approved by Animal House of Achievers University, Owo, Nigeria. (Ref no: AHAU/IEC/20156/2017) and is in accordance with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. Fifteen rabbits of the same sex, divided into three groups of five rabbits each, were used for the study. The rabbits were bought from an animal farm in Owo, Nigeria, and were presented to the Federal School of Agriculture, Akure, for confirmation. Ethical clearance on the use of animals for research work was obtained from the Department of Biological Sciences, Achievers University, Owo, Nigeria. The groups were classified as follows:

  • Group A – Five control rabbits had been fed with normal meal and water throughout the period of investigation
  • Group B – Five rabbits were given 30 mg/kg body weight (BW) subcutaneous injection of amoxicillin every 24 h for 7 days, followed by 30 ml raw cucumber fruit juice supplementation for 14 days
  • Group C – Five rabbits were given 30 mg/kg BW subcutaneous injection of amoxicillin every 24 h and raw cucumber fruit juice supplementation for 14 days simultaneously.


Amoxicillin was bought from a registered pharmaceutical shop in Owo, Nigeria. Overdose of 30 mg/kg BW subcutaneous injection of amoxicillin was given every 24 h for 7 days.

Cucumber (Cucumis sativus) was bought from fruit vendors in Owo, Nigeria. The fruit was presented to the 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 a daily basis. A freshly prepared juice was used throughout the period of investigation.

Five milliliters of venous blood was collected from each of the rabbits into lithium heparinized bottles for biochemical analysis after 14 days. All serum parameters were determined by using RANDOX kit.

Plasma transferrin sample is reacted with a buffer containing antibody specific for human transferrin (siderophilin). The absorbance (600 nm) of the resulting turbid solution is proportional to the concentration of transferrin in the sample. By constructing a standard curve from the absorbance of standards, transferrin concentration of sample can be determined.

Plasma Fe3+ is dissociated from its carrier protein, transferrin, in an acid medium and simultaneously reduced to the ferrous form. The ferrous iron is then complexed with the chromogen, a sensitive iron indicator, to produce a blue chromophore which absorbs maximally at 595 nm.

Plasma AST was also determined. This method is an optimized standard method according to the concentrations recommended by the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC). α-oxoglutarate reacts with L-aspartate in the presence of AST to form L-glutamate plus oxaloacetate. The indicator reaction utilizes the oxaloacetate for a kinetic determination of nicotinamide-adenine dinucleotide (NADH) consumption.

Plasma ALT catalyzes the transamination of L-alanine to α-ketoglutarate, forming L-glutamate and pyruvate. The pyruvate formed is reduced to lactate by lactate dehydrogenase with simultaneous oxidation of reduced NADH. The change in absorbance is directly proportional to the ALT activity and is measured using a bichromatic (340, 700 nm) rate technique. All plasma transferrin, Fe3+, ALT, and AST levels were analyzed after 14 days.

Statistical analysis

Information from this study was subjected to statistical analysis to determine mean and standard deviation using SPSS 18.0 software (IBM Corp, Armonk, New York, USA) with Students “t” test and probability value at 0.05 level of significance.


  Results Top


The results obtained showed a significant increase in plasma ALT, AST, and iron and a significant decrease in plasma transferrin when the rabbits were given 30 mg/kg BW subcutaneous injection of amoxicillin compared with the results obtained from their basal samples and control rabbits and when they were given 30 ml of raw cucumber fruit juice supplementation for 14 days with P < 0.05 [Table 1], [Table 2] and [Figure 1].
Table 1: Mean and standard deviation of plasma values of alanine transaminase (U/L), aspartate transaminase (U/L), plasma iron (μg/dL), and transferrin (mg/dL) obtained from the rabbits

Click here to view
Table 2: Comparative analysis of the mean and standard deviation of plasma values of plasma iron (μg/dL), transferrin mg/dL), alanine transaminase/serum glutamate pyruvate transaminase (U/L), and aspartate transaminase/ serum glutamic oxaloacetic transaminase (U/L) obtained from the rabbits

Click here to view
Figure 1: Comparative analysis of the mean and standard deviation of plasma values of plasma iron/μg/dL, transferrin/mg/dL, alanine transaminase/serum glutamate pyruvate transaminase/U/L, and aspartate transaminase/serum glutamic oxaloacetic transaminase/U/L obtained from the rabbits

Click here to view


There was also a significant increase in plasma AST when the rabbits were supplemented with 30 ml of cucumber fruit juice (P < 0.05) [Table 1], [Table 2] and [Figure 1].

The results obtained showed a significant decrease in plasma ALT, AST (except when the result was compared with cucumber juice supplementation), and iron and a significant increase in plasma transferrin when the rabbits were given 30 ml of raw cucumber fruit juice for 14 days than when they were given 30 mg/kg BW subcutaneous injection of amoxicillin every 24 h for 7 days with P < 0.05 [Table 1], [Table 2] and [Figure 1].

The results obtained showed no significant changes in plasma ALT, AST, iron, and transferrin when the rabbits were given 30 ml of raw cucumber fruit juice for 14 days compared with the results obtained from their basal samples and control rabbits and when they were given 30 ml of raw cucumber fruit juice supplementation with P > 0.05 [Table 1], [Table 2] and [Figure 1].

The results obtained showed no significant changes in plasma ALT, AST, iron, and transferrin when the rabbits were given 30 mg/kg BW subcutaneous injection of amoxicillin every 24 h and 30 ml raw cucumber fruit juice supplementation for 14 days simultaneously with P > 0.05 [Table 1], [Table 2] and [Figure 1].


  Discussion Top


This work was used to assess the plasma iron, transferrin alanine, and AST in amoxicillin overdose supplemented with raw cucumber juice.

The results obtained showed a significant increase in plasma ALT, AST, and iron and a significant decrease in plasma transferrin when the rabbits were given 30 mg/kg BW subcutaneous injection of amoxicillin compared with the results obtained from their basal samples and control rabbits and when they were given 30 ml of raw cucumber fruit juice supplementation for 14 days. This significant increase in plasma ALT, AST, and iron and a decrease in plasma transferrin could be due to amoxicillin-induced hepatotoxicity and inflammation as ALT is found in plasma and in various body tissues, but it is most common in the liver. The plasma level increases in hepatotoxicity and hepatitis as ALT leaks from the liver to the plasma.[20] In addition, AST and iron are found in large quantity on red blood cells and liver.[9],[23] Upon hemolysis, hepatotoxicity, hepatocellular damage, and inflammation[9] which could be drug induced, the plasma levels of these parameters will be abnormally raised, which is attributable to the findings of this work as amoxicillin overdose could result into hemolysis, hepatotoxicity, hepatocellular damage, and inflammation[9] because the side effects include black, tarry stools; bleeding gums; blood in the urine; bloody nose; dark urine; bloody stool; fever; heavier menstrual periods; inflammation; nosebleeds; pain; burning while urinating; painful or difficult urination; pale skin, ulcers, or white spots in the mouth; sudden decrease in the amount of urine; unusual bleeding or bruising; unusual tiredness or weakness;[24] unusual weight loss; vomiting of blood; watery or bloody diarrhea; and jaundice-yellow eyes or skin.[25],[26],[27] Decrease in plasma transferrin could be linked with the physiological role of transferrin, leading to the excessive use of the transferrin to bind with excess plasma iron as a result of hepatotoxicity, inflammation, hemolysis, and hepatocellular damage because transferrin is an iron-binding blood plasma glycoprotein that controls the level of Fe in body fluids.[21]

The results obtained also showed a significant decrease in plasma ALT, AST, and iron and a significant increase in plasma transferrin when the rabbits were given 30 ml of raw cucumber fruit juice for 14 days than when they were given 30 mg/kg BW subcutaneous injection of amoxicillin every 24 h. These findings could be attributed to the health benefit bioactivities of raw cucumber fruit juice (Cucumis sativus) such as antioxidant bio-activity to prevent cellular damage and cleansing action which include removal of accumulated waste materials and chemical toxins from the body.[28] There was also a significant increase in plasma AST when the rabbits were supplemented with 30 ml of cucumber fruit juice. This could be explained as the juice might not be enough to reverse elevated AST or may have hemolytic effect or might have caused myocardial infarction or kidney damage as AST is also found in kidney, red blood cells, and heart.[29]


  Conclusion Top


This work revealed the cyto- and hepato-protective effects of cucumber fruit juice supplementation as there were no significant changes when the rabbits were given subcutaneous injection of amoxicillin and raw cucumber fruit juice supplementation simultaneously. The findings also include a significant increase in plasma ALT, AST, and iron and a significant decrease in plasma transferrin in amoxicillin overdose, while a significant decrease in plasma ALT, AST, and iron and a significant increase in plasma transferrin when the rabbits were given three supplements of raw cucumber fruit juice. There was also a significant increase in plasma AST when the rabbits were supplemented with 30 ml of cucumber fruit juice. Cucumber fruit juice could therefore be an antidote to hepatotoxicity, while AST measurement should be further evaluated in administering the supplement as it may cause increased AST. Further research should be carried out on cucumber fruit juice toxicity.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
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