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 Table of Contents  
ORIGINAL ARTICLE
Year : 2019  |  Volume : 4  |  Issue : 1  |  Page : 11-16

Biochemical alterations in plasma total bile acid, creatinine, sodium, potassium, chloride, and bicarbonate in rabbits overdosed with ibuprofen and supplemented with guava leaf (Psidium guajava) extracts


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

Date of Submission03-Sep-2018
Date of Acceptance07-Dec-2018
Date of Web Publication20-Jun-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/bjhs.bjhs_24_18

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  Abstract 

STUDY BACKGROUND: Ibuprofen is an analgesic and a nonsteroidal anti-inflammatory drug while Psidium guajava leaf has antioxidative, antimicrobial, anti-inflammatory, and analgesic activities.
AIM AND OBJECTIVE: The aim of the study is to evaluate plasma total bile acids (TBAs), creatinine, and electrolytes in rabbits overdosed with ibuprofen and supplemented with guava leaf (P. guajava) extracts.
JUSTIFICATION: There exists a little information on the change in the plasma levels of TBA and electrolytes in ibuprofen toxicity supplemented with guava leaf extract.
MATERIALS AND METHODS: Fifteen rabbits (weight – 0.8–1.4 kg; age – 1.5 ± 0.4 years, n = 5/group) were grouped into control (a); rabbits administered 1600 mg/KgBW of ibuprofen (B1) before being fed with 400 mg/KgBW of guava leaf aqueous extract (B2) and rabbits administered 1600 mg/KgBW of ibuprofen administration (C1) before being fed with 400 mg/KgBW of guava leaf methanolic extract (C2). Plasma creatinine, sodium, potassium, chloride, and bicarbonate levels were determined using COBAS C111 chemistry autoanalyzer while TBA was determined by spectrophotometry.
RESULTS: The results showed a significant increase in the plasma level of TBA, potassium, and creatinine in the rabbits following the administration of 1600 mg/KgBW of ibuprofen (P < 0.05) and a significant decrease in the plasma TBA, potassium, and creatinine in the same rabbits when they were given 400 mg/KgBW of guava leaf extract following ibuprofen overdose (P < 0.05).
CONCLUSION: There was a significant increase in the plasma values of TBA, creatinine, and potassium following the administration of ibuprofen possibly due to ibuprofen toxicity which was significantly reduced upon the administration of 400 mg/KgBW of guava leaf extract.

Keywords: Creatinine, electrolytes, guava leaf, ibuprofen, total bile acids


How to cite this article:
Olaniyan MF, Okunola PO. Biochemical alterations in plasma total bile acid, creatinine, sodium, potassium, chloride, and bicarbonate in rabbits overdosed with ibuprofen and supplemented with guava leaf (Psidium guajava) extracts. BLDE Univ J Health Sci 2019;4:11-6

How to cite this URL:
Olaniyan MF, Okunola PO. Biochemical alterations in plasma total bile acid, creatinine, sodium, potassium, chloride, and bicarbonate in rabbits overdosed with ibuprofen and supplemented with guava leaf (Psidium guajava) extracts. BLDE Univ J Health Sci [serial online] 2019 [cited 2019 Sep 22];4:11-6. Available from: http://www.bldeujournalhs.in/text.asp?2019/4/1/11/260734

Ibuprofen is a nonsteroidal anti-inflammatory drug used for the treatment of pain, painful menstrual periods, migraines, rheumatoid arthritis, and fever. An abuse of the drug, especially in overdose and prolonged use, may cause ulcers, gastrointestinal bleeding, or holes in the stomach or intestine; heart attack or a stroke; and increased risk of heart failure, kidney failure, and liver failure. It is a common pain relief drug in Nigeria where it is taken by people without medical prescription which can result to abuse.[1] It acts as an anti-inflammatory agent by decreasing the activity of cyclooxygenase enzymes, especially oxygenation of arachidonic acid thereby inhibiting the production of prostaglandins. Prostaglandins play a major role in inflammatory response. They can promote and resolve inflammation.[2]

Psidium guajava is a tropical plant of which the young leaf is applied in traditional medicine to treat diarrhea, lower cholesterol, and preventing diabetes. P. guajava leaf has antimicrobial, anti-inflammatory, and analgesic properties. The leaf extracts contain health-promoting antioxidants. The leaf extract of P. guajava is rich in tannins, phenols, triterpenes, flavonoids, essential oils, saponins, carotenoids, lectins, quercetin, vitamins (e.g., Vitamin C), and fiber and fatty acids making it pharmacological relevant.[3]

Total bile acids (TBA) are steroid acids found in the bile of mammals synthesized in the liver (primary bile acids) and as a result of bacterial actions in the colon (secondary bile acids). Plasma TBA is a good index of liver disease.[4] Electrolytes are minerals in blood and other body fluids that carry an electric charge. Electrolytes as cations and anions include chloride, potassium, sodium, and bicarbonate are responsible for volume of water in the body, blood pH, and muscle function.[4] Electrolytes are water-soluble salts, acids, and bases.[4] Plasma levels of electrolytes are good indications of water-electrolyte and acid-base imbalance in the body. Creatinine is formed from creatine. Plasma creatinine is an indicator of renal dysfunction.[4]

Alterations in plasma TBA, creatinine, sodium, potassium, chloride, and bicarbonate could be as a result of dehydration, overhydration, infections, drug toxicity, liver, and renal diseases.[4]

This work is, therefore, designed to determine possible biochemical alterations in plasma TBA, creatinine, sodium, potassium, chloride, and bicarbonate in rabbits overdosed with ibuprofen and supplemented with young leaf extract of guava (P. guajava).


  Materials and Methods Top


Materials

Study area

This experimental study was carried out at the Achievers University Animal House, Owo Local Government Area of Ondo State in Nigeria. Achievers University is in Owo, Ondo State, Nigeria (NUC, 2013).

Study population

Fifteen rabbits weighing 0.8–1.4 kg and aged 1.5 ± 0.4 years were purchased in Owo and were identified and confirmed having the same sex (male) in the Department of Biological Sciences, Achievers University, Owo, Ondo state, Nigeria. Rabbits with the same sex (male) which were divided into three groups were used in the experiment. Fifteen apparently healthy rabbits were bought and classified into three groups of five rabbits each. Body weight of animals before and after the experiment was measured using Mettler sensitive balance (number 202845).

Study design

The study design was experimental and observational study which design involves 15 rabbits which were divided into three[3] groups: A, B and C; Group A were fed with normal meal for 14 days (control group); Group B were overdosed with 1600 mg/KgBW of ibuprofen for 7 days orally (B1) followed by the administration of 400 mg/KgBW of aqueous extracts of young guava leaf for another 14 days (B2); and Group C were overdosed with 1600 mg/KgBW of ibuprofen for 7 days orally (C1) followed by the administration of 400 mg/KgBW of methanolic extracts of young guava leaf for another 14 days (C2).

The animals were kept for 14 days for acclimatization under standard conditions of temperature and relative humidity with both light and dark cycles before any treatment.

Duration of study

The rabbits were acclimatized for 14 days while the study was carried out for a period of 21 days.

Collection of blood sample

Blood samples were collected after every treatment. Fasting samples were obtained from each of the rabbits for plasma TBA. The samples were preserved in lithium heparinized bottles.

  1. B1 – blood collection after rabbits were given 1600 mg/KgBW of ibuprofen for 7 days
  2. B2 – blood collection after rabbits were given 400 mg/KgBW of aqueous extracts of guava leaf for 14 days following the administration of 1600 mg/KgBW of ibuprofen for 7 days
  3. C1 – blood collection after rabbits were given 1600 mg/KgBW of ibuprofen for 7 days
  4. C2 – blood collection after rabbits were given 400 mg/KgBW of methanolic extracts of guava leaf for 14 days following the administration of 1600 mg/KgBW of ibuprofen for 7 days.


Methods

Treatment of normal control and experimental rabbits

  • Group A: Five rabbits fed with normal meal for 21 days (normal control)
  • Group B: Five rabbits fed with normal meal for 7 days and overdosed with 1600 mg/KgBW of ibuprofen for 7 days (B1) and were treated with 400 mg/KgBW of aqueous extracts of guava leaf for 14 days (B2). Each of their weight was documented
  • Group C: Five rabbits fed with normal meal for 7 days and overdosed with 1600 mg/KgBW of ibuprofen for 7 days (C1) and were treated with 400 mg/KgBW of methanolic extracts of guava leaf for 14 days (C2). Each of their weight was documented.


Ethical consideration

Approval for this study was obtained from the biomedical research ethics committee of the animal house, Department of Biological Sciences, Achievers University, Owo, and was carried out in strict compliance with the guidelines for the care and use of animals for research.

Collection of guava leaves

The fresh and tender local guava leaves were collected and were presented to the Department of Biological Sciences, Achievers University, Owo, for confirmation and certification (AUO/BS/2018/176). It was certified (AUO/BS/2018/176) and confirmed on April 14, 2018, before the commencement of the work.

Preparation of guava leaf extracts

The leaves were cleaned. They were dried for 6–7 days without being exposed to the sunlight at room temperature. Dried leaves were ground using electric grinder. The fine particles were sieved out using fine plastic sieve. The coarse particle was reground and sieved for fine particles. Fifty grams of the fine particles was dissolved in 350 ml methanol and distilled water in separate containers. These preparations were left at room temperature for the extraction. The extracts from these were filtered, concentrated after dryness using an evaporator at 45°C. Four hundred milligram of the extract powder was dissolved in 2 mL of distilled water for administration.

Water and methanol (common solvents used in extracting active phytochemicals and phytonutrients from plants) were used to determine their effectiveness and the hazardous effect on the body system.

Preparation of ibuprofen powder

Ibuprofen tablets were purchased and ground into powder using laboratory pestle and mortal.

Blood sample preparation

Whole blood sample was collected from the veins lining the ear of each of the rabbits by the use of 2 mL syringe and was dispensed into sterile vacutainer bottle containing lithium heparin anticoagulant and gently mixed by inverting the container several times for the determination of renal function markers. Plasma was separated from the blood by centrifugation using bench/macro centrifuge for 5 min at 4000 rpm, into plain bottles, and stored at − 20°C until time of analysis.

Estimation of plasma creatinine, sodium, potassium, chloride, and bicarbonate

Estimation of plasma creatinine, (05401755190) sodium (21029371001), potassium (04834925001), chloride (04834909001), and bicarbonate (03289923190) were carried out by autochemistry analysis using COBAS C111 and Roche reagent.

Estimation of total bile acid by enzymatic colorimetric method using reagent kit of Randox

Principle

In the presence of thio-nicotinamide-adenine dinucleotide (NAD), the enzyme 3-alpha-hydroxysteroid dehydrogenase (3-α HSD) converts bile acids to 3-ketosteroids and thio-NADH. The reaction is reversible and 3-α HSD can convert 3-ketosteroids and thio-NADH to bile acids and thio-NAD. In the presence of excess NADH, the enzyme cycling occurs efficiently, and the rate of formation of thio-NADH is determined by measuring specific change of absorbance at 405 nm.

Statistical analysis of data

The results obtained from this study were subjected to statistical analysis using the Statistical Package for the Social Science 18.0 (IBM SPSS 18.0 Armonk, New York, United States) to determine, mean, standard deviation, Students t-test and probability at 0.05 level of significance.


  Results Top


The result [Table 1], [Table 2] and [Figure 1], [Figure 2] obtained showed a significant increase (P< 0.05) in plasma TBA (B1: 11.2 ± 1.0 μmol/L and C1: 12 ± 1.5 μmol/L), potassium (B1: 9.1 ± 1.0 mmol/L and C1: 9.2 ± 1.0 mmol/L), and creatinine (B1: 4.0 ± 0.5 mg/dl and C1: 3.8 ± 0.4 mg/dl) levels in rabbits given 1600 mg/KgBW of ibuprofen per oral for 7 days compared with the results of the control rabbits (TBA: 4.1 ± 0.2 μmol/L; creatinine: 1.0 ± 0.2 mg/dl; and K: 3.4 ± 0.5 mmol/L).
Table 1: The mean (μ) and standard deviation (σ) of plasma sodium, potassium, chloride, bicarbonate, and creatinine obtained in the rabbits

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Table 2: The comparative analysis of mean (μ) and standard deviation (σ) of sodium, potassium, chloride, bicarbonate, and creatinine obtained in test and control rabbits

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Figure 1: The comparative description of mean (μ) and standard deviation (σ) of Na, Cl, and bicarbonate obtained in test and control rabbits

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Figure 2: The comparative description of mean (μ) and standard deviation (σ) of total bile acids, K, and creatinine obtained in test and control rabbits

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The result [Table 1], [Table 2] and [Figure 1], [Figure 2] also showed a statistically significant (P< 0.05) decrease in plasma level of TBA (B2: 4.2 ± 0.3 μmol/L and C2: 4.1 ± 0.3 μmol/L), potassium (B2: 4.2 ± 1.0 mmol/L and C2: 4.0 ± 1.01.0 mmol/L), and creatinine (B2: 1.8 ± 0.3 mg/dl and C2:1.8 ± 0.3 mmol/L) in the rabbits when they were given 400 mg/KgBW of aqueous and methanolic extract of guava leaf daily for 14 days after 7 days of post ibuprofen administration compared to when the same rabbits were given 1600 mg/KgBW of ibuprofen per oral for 7 days (Groups B1 and C1).

However, there was no significant difference in plasma levels of sodium, bicarbonate, and chloride [Table 1], [Table 2] and [Figure 1], [Figure 2] in rabbits given 1600 mg/KgBW of ibuprofen per oral, for 7 days (Groups B1 and B2) compared with the results obtained in the control rabbits (Group A).

Furthermore, the result showed no significant difference in plasma level of TBA, sodium, bicarbonate, and chloride [Table 1], [Table 2] and [Figure 1], [Figure 2] in rabbits given aqueous and methanolic extract of guava leaves for 14 days after 7 days of post ibuprofen administration (Group B2 and Group C2) compared to when the same rabbits were given 1600 mg/KgBW of ibuprofen per oral for 7 days (Group B1 and Group C1).

The result also showed no significant difference in plasma level of TBA, creatinine, sodium, bicarbonate, potassium, and chloride [Table 1], [Table 2] and [Figure 1], [Figure 2] in rabbits treated with aqueous guava extract daily for 14 days after 7 days of postibuprofen inducement (Group B2) compared to when the rabbits were administered with the methanolic extract (Group C2).


  Discussion Top


The result obtained in this study showed a significant increase in potassium and creatinine levels. The result is in line with the findings of Machado et al. that increase in creatinine and potassium levels in rabbits overdosed with ibuprofen is due to its toxic effect on the kidney as ibuprofen overdose has a renal side effects such as urinary retention, renal insufficiency, acute renal failure, nephrotic syndrome, and acute tubular necrosis.[1] Ibuprofen nephrotoxicity (renal toxicity) interferes with the renal effects of prostaglandins, thus reducing blood flow to the kidneys and potentially decreasing renal function. These are the findings when the body is exposed to drug or toxin that can cause kidney damage. Whenever there is kidney damage, the organ will not be able to excrete urine, wastes products of metabolism, and blood electrolytes (such as potassium, sodium, chloride, and bicarbonate) as they will accumulate and their plasma level will beget elevated.[4] Nonsteroidal anti-inflammatory drugs can cause different types of kidney damage including acute kidney injury; water-electrolyte and acid-base imbalance; acute interstitial nephritis and nephrotic syndrome; and papillary necrosis.[4],[5] Nonsteroidal anti-inflammatory drugs can inhibit the synthesis of prostaglandin, to produce different complications such as elevated creatinine levels, hyperkalemia, hyponatremia, and edema[4],[6] as a result of kidney dysfunction which can be reversed when the therapy is discontinued. The increased plasma level of creatinine can indicate kidney damage caused by ibuprofen overdose as a result of possible renal toxicity in experimental rabbits. The hyperkalemia obtained in the experimental animal is an evidence of ibuprofen driven renal damage.

The result also showed a significant decrease in plasma level of potassium and creatinine. It has been reported that P. guajava leaf extract is very rich in tannins, antioxidants (such as phenols, triterpenes, flavonoids, essential oils, saponins, carotenoids, lectins, and vitamins), and fiber and fatty acids which make guava leaves pharmacological significant to have health-promoting therapeutic properties.[3] The decreased level of TBA, creatinine, and potassium in rabbits administered with guava leaf extract shows possible hepatoprotective and nephron-protective effects of guava leaf extracts in ibuprofen toxicity. The result is in accordance with the findings of Ryu et al.[7] that guava leaf extract has therapeutic and health-promoting properties such as an analgesic, anti-inflammatory, antimicrobial, and antioxidant which may be due to the presence of phenolic compounds.[3] Increase in TBAs following ibuprofen overdose could be attributed to the fact that bile acids are synthesized and found in the liver which as a result of liver disease/hepatotoxicity is released into the plasma, thereby causing a significant increase in TBA.[8],[9],[10] This could be attributed to the findings of this work as the plasma level of plasma TBA increased significantly following ibuprofen overdose possibly due to ibuprofen-induced hepatotoxicity.[8],[9],[10] Significant increase in plasma creatinine and potassium after an overdose of ibuprofen could be possible drug-induced nephrotoxicity and hemolysis caused by ibuprofen overdose.[4]

In this study, rabbits treated with guava leaf extract (both aqueous and methanolic) showed reduction in ibuprofen toxicity as reflected by decrease in plasma value of TBA, potassium, and creatinine as against the result obtained in ibuprofen overdose.


  Conclusion Top


This work revealed a significant increase in plasma level of potassium and creatinine in experimental rabbits following the administration of 1600 mg/KgBW of ibuprofen. The biochemical change in plasma value of potassium and creatinine was, however, reversed upon the administration of 400 mg/KgBW of guava leaf extracts (both aqueous and methanolic) possibly due to its phytochemicals component.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

 
  References Top

1.
Machado GC, Maher CG, Ferreira PH, Day RO, Pinheiro MB, Ferreira ML. Non steroidal anti-inflammatory drugs for spinal pain: A systematic review and meta-analysis. Pharm J 2017;15:25-7.  Back to cited text no. 1
    
2.
Ricciotti E, FitzGerald GA. Prostaglandins and inflammation. Arterioscler Thromb Vasc Biol 2011;31:986-1000.  Back to cited text no. 2
    
3.
Gutiérrez RM, Mitchell S, Solis RV. Psidium guajava: A review of its traditional uses, phytochemistry and pharmacology. J Ethnopharmacol 2008;117:1-27.  Back to cited text no. 3
    
4.
Burtis CA, Bruns DE. Tietz Fundamentals of Clinical Chemistry: Molecular Diagnostics (Fundamentals of Clinical Chemistry (Tietz). 7th ed. St. Louis: Elsevier/Saunders; 2015.  Back to cited text no. 4
    
5.
Gooch K, Culleton BF, Manns BJ, Zhang J, Alfonso H, Tonelli M, et al. NSAID use and progression of chronic kidney disease. Am J Med 2007;120:280.e1-7.  Back to cited text no. 5
    
6.
Howard TE. Clinical Chemistry. New York: John Wiley and Sons; 1989. p. 4, 58-62.  Back to cited text no. 6
    
7.
Ryu NH, Park KR, Kim SM, Yun HM, Nam D, Lee SG, et al. Ahexane fraction of guava leaves (Psidium guajava L.) induces anticancer activity by suppressing AKT/mammalian target of rapamycin/ribosomal p70 S6 kinase in human prostate cancer cells. J Med Food 2012;15:231-41.  Back to cited text no. 7
    
8.
Hofmann AF, Hagey LR, Krasowski MD. Bile salts of vertebrates: Structural variation and possible evolutionary significance. J Lipid Res 2010;51:226-46.  Back to cited text no. 8
    
9.
Russell DW. The enzymes, regulation, and genetics of bile acid synthesis. Annu Rev Biochem 2003;72:137-74.  Back to cited text no. 9
    
10.
Chiang JY. Bile acids: Regulation of synthesis. J Lipid Res 2009;50:1955-66.  Back to cited text no. 10
    


    Figures

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    Tables

  [Table 1], [Table 2]



 

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