Academic Sciences International Journal of Applied Pharmaceutics

ISSN- 0975-7058 Vol 4, Issue 3, 2012

Research Article
ASPIRIN INDUCED CHANGES IN SERUM ACP, ALP, GOT, GPT, BILIRUBIN AND CREATININE IN
CORELATION WITH HISTOPATHOLOGICAL CHANGES IN LIVER AND KIDNEY OF FEMALE
ALBINO RAT

NEHA JAIN, RENU SHRIVASTAVA, ARUN K RAGHUWANSHI AND VINOY K SHRIVASTAVA*

Laboratory of Endocrinology, Department of Biosciences, Barkatullah University, Bhopal 462026 Madhya Pradesh, India.
Email: vinoyks2001@yahoo.com
Received: 06 Jan 2012, Revised and Accepted: 15 Feb 2012
ABSTRACT
The aim of present investigation was to study the effect of oral administration of aspirin drug i.e. acetylsalicylic acid (ASA) on female albino rat,
Rattus norvegicus. The animals (n=24) were allocated into 2 groups as control (n=12) and treated (n=12). The treated rats were administered orally
(through gavage) a dose of aspirin 100 mg/ kg body weight for 15 days (n=6) and 30 days (n=6). The serum biochemical estimations such as ACP,
ALP, GOT, GPT, bilirubin and creatinine levels along with histopathological study of liver and kidney were done. In present investigation, aspirin
significantly elevated serum ACP, ALP, GOT, GPT, creatinine and bilirubin levels in both durations. In connection to this, it induced significant
histopathological variations in liver and kidney after 15 and 30 days. All these results suggest that the high doses of ASA induced histopathological
changes in liver and kidney directly or through modulation, certain biochemical and enzymological activities. These effects are dose and duration
dependent.
Keywords: Aspirin (acetylsalicylic acid, ASA), Biochemical estimations, Histopathological study, Rattus norvegicus.

INTRODUCTION Ecospirin-75 (manufactured by USV LTD, B.S.D. Marg, Govandi,

Mumbai-400088).
Aspirin (acetylsalicylic acid, ASA) has been used as one of the most
famous, cheap, easily available and widely used Non Steroidal Anti Experimental animals
Inflammatory Drug (NSAID). Aspirin is used in versatile purpose
such as, anti-inflammatory (in joint diseases), anti-platelets (in The above study was carried out on 24 adult female albino rats
cardiovascular disease), analgesic and antipyretic1. Salicylic acid is (body weight ranging between 150-200 gm) were kept into
metabolized via conjugation in the liver to form salicyluric acid and polypropylene cages and acclimatized to laboratory condition at 23-
25ºC temperature with 14 hours light and 10 hours dark cycle at
several other metabolites. It is also well known that, aspirin is
least for 7 days prior to initiating to the experiment. The study was
rapidly absorbed from the stomach and small intestine, primarily by
conducted as per guidelines of Committee for the purpose of control
passive diffusion across the gastrointestinal (GI) tract which rapidly
and supervision on experiments on animals (CPCSEA), Chennai and
hydrolyzed to salicylic acid by esterase in the GI mucosa and blood
maintained in the Laboratory of Bioscience, Barkatullah University.
plasma. It dispersed throughout the body after ingestion, with the
The animals were fed with standard rat feed and water ad libitum. A
highest concentrations found in the blood plasma, liver, renal cortex,
dose of aspirin (100mg/kg b. wt.) was daily administered orally,
heart and lungs2. through gavage (administration of food or drugs by force, especially
Aspirin is a safe drug at low doses but also it has life-threatening to an animal) for 15 and 30 days. The animals were divided into 3
side effects when administered at high doses. Long-term therapeutic groups of 6 each.
administration of aspirin is associated with nephrotoxicity, Group 1: Fed with normal diet and water ad libitum served as
hepatotoxicity, gastrointestinal ulcerations, and even renal cell control.
cancer due to its adverse effects on multiple organ systems3,4. In
connection to this also reported that, it can cause adverse affects in Group 2: Fed with normal diet and given aspirin orally (100mg/kg
pregnancy5. In-vitro and in-vivo studies showed that aspirin at high b. wt.) through gavage for 15 days.
doses caused death of the blood vessel tissues6. The inhibitory
Group 3: Fed with normal diet and given aspirin orally (100mg/kg
activity of aspirin also found on the endocrine hormones viz. ACTH,
b. wt.) through gavage for 30 days.
endorphin, cortisol, prolactin and growth hormone via possible
stimulatory role of prostaglandin. Also, overdose of aspirin stimulate After above treatments, the animals were weighed, sacrificed on 16th
corticosteroid secretion by the adrenal cortex4. and 31st day, blood samples were collected through cardiac
puncture, serum were separated for enzymological and biochemical
Aim of the work estimations and organs like liver and kidney were preserved in
The aim of the present study is to observe the side effects of the anti- Bouin’s fluid for histopathological observations. Therefore, following
inflammatory drug aspirin (100 mg/ kg body weight) on liver and parameters were done by appropriate methods.
kidneys of young albino female rats, Rattus norvegicus, by observing Parameters
histopathological studies in liver and kidney and by estimating
enzymological i.e. ACP (Acid Phosphatase), ALP (Alkaline Serological analysis
Phosphatase), GOT (Glutamate Oxaloacetate Transaminase), GPT
(Glutamate Pyruvate Transaminase) and certain biochemical i.e. 1. King and Kings method was used for the determination of
bilirubin and creatinine levels in serum after 15 and 30 days of the serum Acid Phosphatase (ACP) and serum Alkaline
treatments. Phosphatase (ALP)7.

MATERIALS AND METHODS 2. Reitman and Frankel method was used for the determination of
serum Glutamate Oxalate Transaminase (GOT) and serum
Material Glutamate Pyruvate Transaminase (GPT)8.

Aspirin (acetylsalicylic acid) tablets (75 mg/tablet) as anti- 3. Malloy and Evelyn method was used for the determination of
inflammatory drug was purchased from the market, trade name serum Bilirubin9.
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Int J App Pharm, Vol 4, Issue 3, 2012, 9-11

4. Alkaline picrate method was used for the determination of atrophy in liver and kidney via inhibition of the synthesis of
serum Creatinine10. different prostaglandin viz. PGE2, PGD2, PGF2 and PGI2 which are
potent vasodilators1,18.
Histopathological study
In present study, aspirin (100mg/Kg b.wt.) significantly elevated
For histological study liver and kidneys were dissected out quickly, ACP, ALP, GOT, GPT, creatinine and bilirubin content in serum upto
cleaned, dried with blotting paper and fixed in Bouin’s fluid. The 15 and 30 days treatment (Table 1).
classical paraffin sectioning (7µ thick) were cut, stained with
Haematoxylin and Eosin staining and observed under light And, the administration of aspirin caused significant histo-
microscopy for histopathological changes11. pathological changes in liver and kidney (Fig. 1-6). Aspirin treated
liver of rat showed degenerative and pycknotic changes in the nuclei
Statistical analysis with less cytoplasmic materials having dense particles in the
hepatocytes. As well as, vacuolization, clear dilations were also
The ‘p’ value of treated v/s control were calculated by adopting
found in the sinusoids, most of the hepatocytes become
Student‘t’-test12.
hypertrophied in condition with small and degenerating nuclei and
RESULT AND DISCUSSION lobules have become fused in later part of the experiment (Fig. 2, 3).
In connection to this, aspirin treated kidney of rat showed atrophic
Aspirin develops hepatotoxicity i.e. lethal hepatocellular injury and changes in the proximal and distal convoluted tubules in the cortex,
hepatic massive micro-steatosis via mitochondrial dysfunction and the inflammatory changes were noticed in medulla and damaged
lipid peroxidation mechanism resulting marked fall in intracellular tubular epithelial cells by ischemia due to vasoconstriction of renal
ATP and disrupt free fatty acid accumulation in liver 13,14. Aspirin arterioles (Fig. 5, 6). Renal vascular tone was determined by
also develops nephrotoxicity i.e. vacuolar degeneration of tubules autonomous intrinsic activity of the renal arterioles and continuous
and gradually elevates membrane associated phosphatase (ACP and production of prostaglandin which is inhibited by this drug and thus
ALP) and transaminase (GOT and GPT) enzymes of tissues, these caused unopposed constriction of arterioles resulting in ischemia of
enzymes maintain the amino acid homeostasis15,16,17,. It also tubules and epithelial cell death19. Other workers also suggested that
increases serum bilirubin and creatinine content in aspirin treated aspirin caused vacuolar degeneration of proximal tubules, focal
patients due to hepatic and renal toxicity15. Due to, aspirin tubular atrophy and significantly decreased proximal tubules per
administration mainly induced vasoconstriction and smooth muscle unit area, resulting renal failure 20,21.

Table 1: Effect of aspirin on serum bio-chemical constituents of rat, Rattus norvegicus.

Parameters (mg/dl) Control 15 days treated group 30 days treated group
ACP 9.23 ± 0.44 13.44 ± 3.58*** 14.59 ± 0.28***
ALP 10.37 ± 0.63 12.56 ± 1.01*** 15.58 ± 0.60***
GOT 7.33 ± 0.63 16.58 ± 0.56*** 20.16 ± 0.63***
GPT 6.55 ± 1.33 14.55 ± 1.33*** 15.55 ± 1.33***
Creatinine 0.55 ± 0.44 2.18 ± 0.33*** 4.33 ± 0.63***
Bilirubin 0.11 ± 0.06 0.24 ± 0.06*** 0.23 ± 0.06***
Mean ± SD, n = 6, ***p< 0.001(Highly significant)

1 2 3

4 5 6

10
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Int J App Pharm, Vol 4, Issue 3, 2012, 9-11

Explanation of figures of Liver and Kidney case-control study in general practice research database, BMJ,
2000; 320, p1642-1646.
Fig 1. Section of liver of control Rattus norvegicus showing normal 4. Luigi DL, Laura G, Francesco R, Carlo B, and Domenico C,
features of hepatocytes having prominent and spherical nuclei with Aspirin, exercise and pituitary hormones, Official Journal of the
defined cytoplasmic materials (H & E 400X). American College of Sports Medicine, 2001; p2029-2035.
Fig 2. Aspirin treated (100 mg/kg) liver of Rattus norvegicus upto 15 5. Collins E and Turner G, Maternal Fetal effects of regular
days showing hypertrophied and degenerative hepatocytes with salicylate ingestion during pregnancy, Lancet, 1975; 2, p335–
pycknotic changes in the nuclei having less cytoplasmic materials (H 339.
& E 400X). 6. Starke RD, Smith SC, Blair SN, Bonow RO, Brass LM et al.,
AHA/ACC guidelines for preventing heart attack and death in
Fig 3. Aspirin treated (100 mg/kg) liver of Rattus norvegicus upto 30 patients with atherosclerotic cardiovascular disease, The
days showing vacuolization, lobules have become fused, clear American College of Cardiology, 2001; 104, p1577–9.
dilations found in sinusoids, most of the hepatocytes become 7. King PRN, King EJ, Colorimetric method for the determination
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E 400X). Phosphatase, J Clin Path, 1954; 7, p132-136.
8. Reitman S, Frankel SA, Colorimetric method for the
Fig 4. Cortical region of kidney of control Rattus norvegicus showing
determination of Serum Glutamate Pyruvate Transaminase and
normal interstitial connective tissue, peri-tubular capillaries,
Serum Glutamate Oxaloacetate Transaminase, Am J Clin Path,
proximal convoluted tubules (PCT) (having prominent brush border,
1957; 28, p56-62.
wide lumen) and distal convoluted tubules (DCT) (having narrow
9. Malloy, Evelyn, Geoffrey ADH and King EJ, A diazotized
lumen) which have prominent and spherical nuclei (H & E 400X).
sulfanilic acid method for the estimation of bilirubin in blood
Fig 5. Aspirin treated (100 mg/kg) cortical region of kidney of plasma, J Soc chemistry, 1937; 56 (97), p123.
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and DCT but vacuolar degeneration found (H & E 400X). 1953; 6, p201.
11. Durury RAB and Wallington EA, Histological technique, 5th Ed,
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levels in serum of young albino female rats, Rattus norvegicus. If it is toxicological interaction between acetylsalicylic acid and
taken longer duration by patients; they may suffer from renal and diazinon in albino Rats, J Egypt Soc Toxicol, 2006; 35, p1-6.
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ACKNOWLEDGEMENT
JRMS, 2010; 17 (1), p43-45.
We express our sincere thanks and gratitude to Prof. VK Shrivastava, 17. Sangeetha B and Krishnakumari S, Tephrosia purpurea (linn.) a
HOD, Bioscience Department Barkatullah University, Bhopal, folk medicinal plant ameliorates carbon tetrachloride induced
Madhya Pradesh, India for providing departmental infrastructural hepatic damage in rats, International Journal of Pharma and
facilities. Bio Sciences, 2010; 1 (2), p1-10.
18. Vane JR, Inhibition of prostaglandin synthesis as a mechanism
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