A study of histopathological effects and functional tests in liver and kidney of laboratory male mice treated with ammonium chloride

Int. J. Biosci. 2016
RESEARCH PAPER
OPEN ACCESS
A study of histopathological effects and functional tests in liver
and kidney of laboratory male mice treated with ammonium
chloride
Ali A.A. Al-Ali * , Faris S. Kata, Majda I. Abd Al-Majeed
Department of Biology, University of Basrah, Iraq
International Journal of Biosciences | IJB |
ISSN: 2220-6655 (Print), 2222-5234 (Online)
http://www.innspub.net
Vol. 8, No. 2, p. 184-194, 2016
Key words: Ammonium chloride, Histopathological Changes, AST, Urea, Triglyceride.
http://dx.doi.org/10.12692/ijb/8.2.184-194 Article published on February 28, 2016
Abstract
The present study was aimed to investigate the effects of ammonium chloride on mice. The animals were divided
into three groups: groups 2 and 3were treated orally with ammonium chloride at doses of 2 and 4 mg/body
weight, respectively for a period of three weeks, while group 1 served as a control group. The tissue sections
which were made from mice organs (liver and kidney) which treated with 4mg/ body weight of ammonium
chloride proved that this dose has toxic effect only on the liver and kidneys. In the liver, the histopathological
effects are represented by hypertrophy and irregular shape of nucleus, degeneration of cytoplasm, congestion of
sinusoid, bleeding and infiltration of inflammatory cells. In kidneys, the effects focus on renal tubules only which
are represented by the degenerative changes, necrosis and infiltration of inflammatory cells. This study was also
carried out to investigate the influence of ammonium chloride on levels of urea, creatinine, total protein, lipid
(triglyceride and cholesterol), and liver marker enzymes such as AST, ALT and ALP. Oral administration of
ammonium chloride (4mg/body weight) caused a significant increase in the levels of AST and a significant
decrease in the level of ALP and total protein in mice. Treated mice with ammonium chloride at a dose of 2 and
4mg/ body weight for 21 days showed a significant decrease in levels of creatinine, triglyceride and cholesterol,
while ALT and urea had no affect at two doses of ammonium chloride. In conclusion, ammonium chloride causes
direct hepatotoxicity and nephrotoxicity.
* Corresponding Author: Ali A.A. Al-Ali qalialali_75@ymail.com
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Int. J. Biosci. 2016
Introduction
Ammonium chloride is a reagent that is used in a
variety of industrial and research applications.
Industrial uses include electroplating, tinning, and
manufacture of dyes. It is also used as a fluxing agent
for the galvanizing of steel, the refinement of zinc and
the coating of sheet iron with zinc (Williams, 2006).
Ammonium chloride acidifies the urine and is used to
dissolve certain type of bladder stones; it may also be
used to promote the excretion of certain toxins or
drugs into the urine. It may be amended with certain
antibiotics such as tetracycline and penicillin to
increase their effect against certain bacteria in the
urine (Foster and Smith Pharmacy, 2010).
In biological research, ammonium is often used for
lyses of human red blood cells (Kang et al., 2002).
Also, it is used in the isolation of proteins from 50S
ribosomal subunits of Bacillus stearothermophilus
(Gewitz, 1987). In mammals, ammonia is an
important nitrogen substrate in several reactions and
plays an important role in nitrogen homeostasis of
mammalian cells. Ammonia is produced by amino
acid and protein catabolism and is toxic to brain and
muscles. Ammonia toxicity results in free- radical
generation that leads to oxidative stress and tissue
damage (Lena and Subramanian, 2004).
Ammonia affects both excitatory and inhibitory
synaptic transmission in the mammalian brain
through a variety of mechanisms (Monfort and
Felipo, 2005). It is the major metabolic end product
during the catabolism of proteins. Because of its high
toxicity, even at a very low concentration in vivo,
ammonia is either excreted directly or converted to
some less toxic compounds such urea, uric acid or
amino acids (Cooper and Plum, 1987; Randall and
Wright, 1987; Campall, 1991; Wood, 1993).
Ammonium is also an endogenous substance that
serves a major role in the maintenance of the acidbase
balance (Hall, 2015).
Ammonia is converted to urea in the liver by ureacycle
enzymes, which is then excreted by the kidneys.
Hyperammonemia may result from genetic defect or
deficiency of the urea-cycle enzymes or from acquired
condition such as Reyes syndrome, liver failure, highdose
chemotherapy and severe infection (Treem,
1994).
It has been well established that in man, rat and dog
urinary ammonia increases in response to metabolic
acidosis (Pollak et al., 1965).
Ammonia toxicity occurs partly via oxidative stress,
which leads to lipid peroxidation and free radical
generation. This causes hepatic dysfunction and
failure, which is a primary cause of neurological
disorders and alteration in the function of central
nervous system associated with hyperammonemia,
such as hepatic encephalopathies, Reye syndrome,
irritability, somnolence, vomiting and derangement
of cerebral function, coma and death (Harikrishnan et
al., 2008; Vijayakumar and Subramanian, 2010).
In the present study, the effects of ammonium
chloride on mice liver and kidney are investigated.
However, no available literatures dealing with
histological changes and function tests in liver and
kidney evoked by ammonium chloride
administration.
Materials and methods
Test material
Ammonium Chloride NH4Cl powder (1kg) product of
(Reagent Chemical Services Company Ltd. UK). This
powder was dissolved in distilled water.
Animals
Forty-eight adult male albino mice Mus musculus L.
weighting 23-25 gm were used for this study and were
obtained from the animals house of Department of
Biology, College of Education, University of Basrah.
Animals were housed in cages (4 animals/ cage) and
were fed with standard diet for 21 days at room
temperature (25±3 o C) with controlled light- dark
cycle throughout the experiment (Jawad, 1996).
Experimental Design
Animals were divided into three groups comprising
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Int. J. Biosci. 2016
eight animals in each group as follows:
Group 1: Mice were served as control treated with 0.2
ml of distal water.
Group 2: Experimental animals were orally given 0.2
ml of ammonium chloride solution at a dose level of 2
mg/ body weight daily, for a period of three weeks.
Group 3: The animals were orally given 0.2 ml
ammonium chloride solution at a dose level of 4 mg/
body weight daily for a period of three weeks.
Histopathological examination
For light microscopic examination, liver and kidney
tissues from each group were fixed with Bouin's
solution and embedded with paraffin. After routine
processing, paraffin sections of each tissue were cut
into 7 μm thickness and stained with hematoxylin and
eosin (Humason, 1972).
Biochemical Analysis
At the end of experimental period, mice were
anaesthetized with ether. Blood samples were
withdrawn by heart puncture and serum was
separated, centrifuged at 3500 rpm for 15 minutes,
and blood serum was then collected and stored at 4 ◦ C
prior immediate determination of serum parameters.
Serum aspartate transaminase (AST) and alanine
transaminase (ALT) were determined according to
the methods of Reitman and Frankel (1957). Serum
alkaline phosphatase (ALP) and urea were
determined according to the methods of King and
King (1954) and Wills and Savory (1981), respectively.
Serum creatinine and total protein were estimated
using the methods of Butch (1994). Serum total
cholesterol and Triglycerides were determined
according to the methods of Tietz (1995). All of these
parameters were measured using spectrophotometer.
Kits were provided by Biolabo and Biomerieux
Company (France).
Statistical Analysis
The results of biochemical tests were analyzed using
the Statistical Package for Social Sciences (SPSS for
windows, version 12.0). Comparisons were made
between experimental groups using one-way analysis
of variance (ANOVA) followed by Dennett's test.
Values of less than 0.01 were regarded as statistically
significant.
Results
Histopathological results
The histopathological examination of the liver of
group 1 and 2 shows normal tissues which included
the center vein, hepatocytes arranged as cords and
between them sinusoids.
Table 1. Effects of ammonium chloride on function tests in liver of mice.
Treatments
AST
(IU/L)
9.63
± 0.41
9.12
± 0.46
*12.21
± 0.89
Control group
(Distal water)
Ammonium chloride
(2mg/ body weight)
Ammonium chloride
(4mg/ body weight)
* Significant differences (p< 0.01 & 0.05) compared with the control group.
ALT
(IU/L)
7.61
± 0.49
6.61
± 0.24
7.92
± 0.23
ALP
(IU/L)
128.61
± 2.51
120.06
± 3.45
*103.15
± 2.74
Histological examination of liver of group 3 (treated
with high dose) revealed histopathological alterations
which included hypertrophy and irregular shape of
nucleus with prominent central nucleoli, massive
degeneration and in other animals, the hepatocytes
had numerous pale vacuoles of widely different sizes.
Other hepatocytes were necrotized. Liver sections
reflected other signs of injury as indicated by
congestion of sinusoid; bleeding and dilatation of
infiltrations by large mass of leukocytic inflammatory
cells were observed. These inflammatory cells include
lymphocytes, giant multinucleated cell and cluster of
epithelioid histiocytes (Fig. 1 and 2).
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Int. J. Biosci. 2016
Table 2. Effect of ammonium chloride on function tests in kidney of mice.
Treatments
Control group
(Distilled water)
Ammonium chloride
(2mg/ day)
Ammonium chloride
(4mg/ body weight)
Urea
(mg/dL)
33.45
± 1.31
32.11
± 1.25
37.38
± 1.47
* Significant differences (p< 0.01 & 0.05) compared with the control group.
Creatinine (mg/dL)
0.95
± 0.03
0.47*
± 0.04
0.34*
± 0.03
Examination of tissue section of kidney of group 3
(treated with high dose) revealed histopathological
changes on lining epithelial tissue of renal tubule
which was represented by degenerative changes,
dilation of lumen of distal convoluted tubule and lysis
of the apical cell membrane of the proximal
convoluted renal tubule and seems that these tubules
have a broad lumen and its lacks the brush boarder in
comparison with the control group. Kidney tissue
sections showed atrophy cells of renal tubule and
deformation of nuclei of other epithelial cells of both
distal and proximal convoluted renal tubule. In
addition, the bleeding was observed between cortical
renal tubule and infiltration of inflammatory cells
near large vessel (Fig. 2 and 3).
Biochemical results
The results of this study showed a significant increase
in aspartate transaminase enzyme level at p

Int. J. Biosci. 2016
Subramanian 2009). By doing so, the liver also plays
a major role in the metabolic regulation of systemic
pH, because hydrogen ions released from NH4 during
the synthesis of neutralize excess bicarbonate
produced by the breakdown of amino acid. An
increase level of circulatory ammonia might indicate a
hyperammonemic condition in rats treated with
ammonium chloride (Lena and Subramanian, 2004;
Essa and Subramanian, 2007). Steven et al. (2009)
showed that the damage of hepatocytes in liver
increased the level of ammonia in blood and caused
hyperammonemic condition due to incapability of
hepatocytes on synthesis of the urea, and all of this
leads to decrease of urea in blood.
Fig. 1. (A) Liver of the control group (400x), (E&H). (B) Liver of treated group (4mg ammonium chloride)
showing hypertrophy of nuclei (arrow) (400x), (E&H). (C) Liver of treated group (4mg ammonium chloride)
showing irregular shape of nucleus in hepatocyte with prominent central nucleoli (arrows) (400x), (E&H). (D)
Liver of treated group (4mg ammonium chloride) showing massive degeneration (thin arrows) and necrosis
(thick arrows) (400x), (E&H). (E) Liver of treated group (4mg ammonium chloride) showing vacuole
degeneration (thin arrows) and congestion of sinusoid (thick arrows) (100x), (E&H). (F) Liver of treated group
(4mg ammonium chloride) showing increase of giant multinucleated cells (arrow) and infiltration (stars) (400x),
(E&H).
The present study the histological examination of
liver and kidney of treated mice with ammonium
chloride showed many histopathological changes
which included hypertrophy and irregular of nucleus,
massive degeneration, necrosis and increase of giant
cells, and congestion of sinusoid in liver. Liver is one
of the most important organs of the body which is
sensitive to toxins being a member of the President
metabolism of many toxic substances entering the
body of the organism. So, it may exhibit severe
histopathological effects than those toxins and exceed
the limits of its ability to get rid of them, which may
explain the diversity of histopathological changes in
the liver and increase the intensity (Klaassen and
Watkins, 1999). The ammonium chloride in mice may
cause changes in the functions responsible for
removing toxins so reflected on the liver's ability to
remove the harmful effects of ammonium chloride
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Int. J. Biosci. 2016
system cells (Rubin and Reisner, 2014). some studies
suggest that exposure to toxins cause a degeneration
of the liver cells to failure in protein representation
and its accumulation (Thophon et al., 2003) or due to
the inhibition of toxic substances the process of
protein synthesis through its effect on the enzyme
protein phosphatase (Stevens et al., 2009). This may
be considered as one of the reasons for the occurrence
of necrosis. The vacuolar degeneration in liver may be
represented by the fatty changes which may be
developed into fatty necrosis (Rubin and Reisner,
2014). This finding is in agreement with Subash and
Subramanian (2009) who reported that 100 mg/kg
body weight; i.p. ammonium chloride rats show liver
fibrosis, steatosis and sinusoidal dilatation. The
gavage studies in rats have shown some lesions of
gastric mucosa with notable histopathological effects
(Takeuchi et al., 1995; Mori et al., 1998). In the
kidney, the pathological changes showed no damage
in the glomeruli and the changes are focused in the
renal tubule, because of the structural nature of the
glomeruli which restrict the movement of blood cells
and most large molecules opposite that allow to many
substances (including water and solutes) in the blood
pass through the renal corpuscle and enter the renal
tubule (Hall, 2015). The histopathological changes of
hyperammoneic mice are infiltration of inflammatory
cells, deformation and hypertrophy of nucleus . The
necrosis of the lining of the renal tubule possibly
causes later renal failure, especially when number of
affected tubule grows (Cotran et al., 1999).
Fig. 2. (G) Liver of treated group (4mg ammonium chloride) showing bleeding and infiltration of inflammatory
cells (200x), (E&H). (H) Liver of treated group (4mg ammonium chloride) showing congestion of the sinusoid
(400x), (E&H). (I) Kidney of control group showing cortical renal tubule (400x), (E&H). (J) Kidney of control
group showing the medullary renal tubules (400x), (E&H). (K) Kidney of treated group (4mg ammonium
chloride) showing deformation of nucleus (400x), (E&H). (L) Degeneration (arrow) and dilation in distal
convoluted renal tubule (stars) of treated group (4mg ammonium chloride) (400x), (E&H).
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Int. J. Biosci. 2016
Oxidative stress injury is actively involved in the
pathogenesis of ammonium chloride and induced
acute kidney injury. Reactive oxygen species (ROS)
directly act on cell components, including lipids,
proteins, and DNA, and destroy their structure.
Increased levels of circulatory ammonia in mice
treated with ammonium chloride may be due to the
liver damage caused by ammonia-induced free radical
generation. Reports have shown that excess ammonia
induces nitric oxide synthase, which lead to the
enhanced production of the nitric oxide, leading in
turn to oxidative stress and liver damage (Kosenko et
al., 2000; Schliess et al., 2002).
Fig. 3. (M) Kidney of treated group (4mg ammonium chloride) showing atrophy in renal cells (1000x) (E&H).
(N) Lysis of apical membrane of proximal convoluted renal tubule cells (arrows) and dilation of lumen (star) of
treated group (4mg ammonium chloride) (400x), (E&H). (P) Bleeding in medullary renal tubules of treated group
(4mg ammonium chloride) (stars) (400x), (E&H). (Q) Kidney of treated group (4mg ammonium chloride)
showing congestion of blood vessel (arrow) and infiltration of inflammatory cells and bleeding between cortical
tubules (star) (400x), (E&H).
The present study showed also a significant increase
in aspartate transaminase enzyme (AST) at high dose
(4 mg ammonium chloride/ day) of treated group
with ammonium chloride, and significant decrease in
alkaline phosphatase with high dose only compared
with control group. The alanine transaminase level in
two treated groups was not affected by treatment as
compared with control (Table 1). The increased
activities of these serum markers observed in this
study correspond to considerable liver damage
induced in ammonium chloride mice. Moreover, the
estimation in serum is useful quantitative marker to
indicate hepatocellular damage (Sallie et al., 1991).
Serum AST, ALT and ALP are the sensitive markers
employed in the diagnosis of liver diseases. When the
liver cell plasma membrane is damaged, numerous
enzymes normally located in the cytosol are released
into the blood stream (Rajesh and Latha, 2004).
Furthermore, the present study showed a significant
decrease in serum creatinine level of mice treated
with both doses (2 and 4mg ammonium chloride) as
shown in table (2), and no significant effect in serum
urea with the two doses, besides a significant decrease
in total protein level with high dose compared with
the control group. The decreasing of serum creatinine
may be due to the decreasing of serum total protein.
The free radicals affect the activity of skeletal muscles
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Int. J. Biosci. 2016
and thereby cause decreasing of serum creatinine
(Kumar et al., 2010). Also, they cause oxidation of
molecular protein in serum and this may result in
decreasing the total protein in serum (Ujjwal and
Dey, 2010). Mostafa et al. (2009) mentioned that the
free radicals caused a significant decrease in plasma
proteins.
Previous reports stated that ammonium (chloride/
acetate) salt induces ammonia toxicity party via
oxidative stress, which leads to lipid peroxidation and
free- radical generation (Essa and Subramanian,
2006). Moreover, the excess of ammonia intoxication
leads to excessive activation of N-methyl-D aspartate
(NMDA) receptors leading to neuronal degeneration
and death (Subash and Subramanian, 2010). Also, the
elevated levels of circulatory liver- markers and lipid
peroxidation products in ammonium chloride in rats
might be due to the liver damage caused by ammoniainduced
free radical generation (Essa and
Subramanian, 2006; Thenmozhi and Subramanian,
2011).
In the present study, the administration of
ammonium chloride caused a significant increase in
the levels of serum lipids (cholesterol and
triglycerides as indicated in Table (3). These findings
indicate that hyperammoemia may be accompanied
by complication of atherosclerosis. Some studies
showed that the levels of serum and tissue lipid were
elevated during hyperammonemic conditions (Lena
and Subramanian, 2004; Lena and Subramanian,
2004; Eassa et al. 2010; Subash and Subramanian,
2012). It has been reported that ammonium
(chloride/acetate) salts may deplete levels of α-KG
and other Krebs cycle intermediates (Lena and
Subramanian; 2004; Essa and Subramanian, 2006)
and thus elevate the levels of acetyl coenzyme A. This
acetyl Co A may be used for the synthesis of fatty
acids and cholesterol, since fatty acids of different
sources are used as substrates for synthesizing
triglycerides and phospholipids. The elevated levels of
acetyl CoA may increase levels of lipid profile (free
fatty acids, triacylglycerols, phospholipids and
cholesterol). Another important function of α-KG
occurs in the formation of carnitine (Vijayakumar and
Subramanian, 2010). Carnitine acts as a carrier of
fatty acids into cell mitochondria so that proper
catabolism of fats can proceed.
Conclusion
In conclusion, ammonium chloride causes direct
hepatotoxicity and nephrotoxicity. The results
indicate that the exposure of male mice to ammonium
chloride affects both liver and kidneys, and this may
be due to the generation of free radicals by showing
their toxicity on liver and kidneys.
Acknowledgement
We would like thank Dr. Farhan Mohsen for his
remarks value in paper output.
References
Campall JW. 1991. Excretory nitrogen metabolism.
In: Prosser, C.L. (Ed.). Environmental and metabolic
animal physiology. Wiley- Liss Inc., New York: 277-
324.
Cooper AJL, Plum F. 1987. Biochemistry and
physiology of brain ammonia. Physiological Reviews
67, 440-519.
http://dx.doi.org/10.1016/j.ab.2015.11.0030003-
2697/©
Cotran RS, Kumar V, Collins T. 1999. Robbins
pathologic basis of diseases 6 th ed., Saunders Co.,
543p.
Essa MM, Ali AA, Waly MI, Guillemin GJ,
Subramanian P. 2010. Effect of leaves on serum
lipids in ammonium chloride induced experimental
hyperammonemic rats. International Journal of
Biologcal and Medical Research 3, 71-73.
Essa MM, Subramanian P. 2006. Pongamia
pinnata modulates oxidant- antioxidant imbalance
during hyperammonemic rats. Fundamental &
Clinical Pharmacology 3, 299-303.
http://dx.doi.org/10.1111/j.14728206.2006.00410.x
191 Al-Ali et al.

Int. J. Biosci. 2016
Essa MM, Subramanian P. 2007. Hibiscus
sabdariffa affects ammonium chloride-induced
hyperammonemic rats, Evidence-Based
Complementary and Alternative Medicine (eCAM)
4(3), 321-325.
http://dx.doi.org/10.1093/ecam/nel087
Foster Smith Pharmacy. 2010. Patient
information sheet.
139-146.
http://dx.doi.org/10.1016/S0008993(00)02785-2
Kosenko E, Kaminsky Y, Lopata O, Muravyov
N, Kaminsky H, Hermenegilioto C, FelipoV.
1998. Nitroarginine, an inhibitor of nitric oxide
syntheses, prevents changes in superoxide radical and
antioxidant enzymes induced by ammonia
intoxication. Metabolic Brain Disease 13, 29-41.
Gewitz HS. 1987. Reconstitution and crystallization
experiments with isolated split proteins from Bacillus
stearothrmophilus ribosome’s. International Journal
of Biochemistry 15(5), 887-895.
Hall J. 2015. Guyton and Hall textbook of medical
physiology. 13 th ed., Saunders, p1043p.
Harikrishnan B, Subramanian P, Subash S.
2008. Antihyperammonemia effect of Withania
somnifera on ammonia induced wistar rat. Cell and
Tissue Research 8, 1417-1420.
Humason GL. 1972. Animal tissue techniques.
3 rd ,Freeman and Company, San Francisco, 641 p.
Jawad AAH. 1996. Ethological studies in assessing
the anti- aggressive effects of some Iraqi medicinal
plant in laboratory mice (Mus musculus). thesis
College of Education, University of Basrah: 204
pp.(In Arabic).
King PR, King EJ. 1954. Estimation of plasma
phosphatase by determination of hydrolysed phenol
with amino-antipyrine. Journal of Clinical Pathology
7, 322-326.
Klaassen CD, Watkins JB. 1999. Casarett and
Doull’s toxicology: The basic science of poisons, 5 th
ed., McGraw Hill, New York: P816 p.
Kosenko E, Kaminsky Y, Stavroskaya IG,
Felipo V. 2000. Alternation of mitochondrial
calcium by ammonia-reduced activation of NMDA
receptor in rat brain in vivo. Brain Research 880,
Kumar R, Malhotra S, Kumar D. 2010.
Antibiotic and free radical scavenging bpotential of
Euphorbia hirta flower extract.
Journal of Pharmaceutical Sciences 72(4), 533-537.
http://dx.doi.org/10.4103/0250-474X.73921
Lena PJ, Subramaninan P. 2004. Effects of
melatonin on the levels of antioxidants and lipid
peroxidation products in the rats treated with
ammonium acetate. Pharmazie 59, 636-639.
Monfort P, Felipo V. 2005. Long-term
potentiation in hippocampus involves sequential
activation of soluble guanylate cycleas, Cgmpdependent
protein kinase and Cgmp-depending
phosphodisterase alteration in hyperammonemia.
BMC. Pharmacology 5(Suppl 1), 66 o.
http://dx.doi.org/10.1186/1471-2210-5-S1-P66
Mori S, Kaneko H, Mitsuma T. 1998.
Implications of gastric topical bioactive peptides in
ammonia-induced acute gastric mucosal lesions in
rats. Scandinavian Journal of Gastroenterology
33(4), 386-393.
http://dx.doi.org/10.1080/00365529850171017
Mostafa A, Saleh M, Bassam A, Samera A.
2009. Oxidative in blood of camels (Camelus
dromedarius) naturally infected with Trypanosoma
evansi. Veterinary Parasitology 162(3-4), 192-199.
http://dx.doi.org/10.1016/j.vetpar.2009.03.035
Nelson DL, Lehninger AL, Cox MM. 2008.
Lehininger principles of Biochemistry. W. H.
Freeman, London: 1100 p.
192 Al-Ali et al.

Int. J. Biosci. 2016
Pollak VE, Mattenheimer EH, De Bruin H,
Weinman KJ. 1965.Experimental metabolic
acidosis. The enzymatic basis of ammonia production
by the dog kidney. Journal of Clinical Investigation
44(2), 169–181.
http://dx.doi.org/10.1172%2FJCI105132
Subash S, Subramanian P. 2009. Morin a
flavonoid exerts antioxidant potential in chronic
hyperammonemic rats: A biochemical and
histopathological study. Molecular and Cellular
Biochemistry 327, 153-161.
http://dx.doi.org/10.1007/s11010-009-0053-1
Rajesh MG, Latha MS. 2004. Preliminary
evaluation of antihepatotoxic activity of kamilari, a
poly herbal formulation. Journal of
Ethnopharmacolog 91(1), 99-104.
http://dx.doi.org/10.1016/j.jep.2003.12.011
Randall DJ, Wright PA. 1987. Ammonia
distribution and excretion in fish.Fish Physiology and
Biochemistry 3, 107-120.
Reitman S, Frankel S. 1957. A colorimetric
method for the determination of sperm GOT and
GPT. American Journal of Clinical Pathology 28, 56-
63.
Rubin E, Reisner HM. 2014. Essential of Rubin's
pathology, 6 th ed., Wolters Kluwer, Lippincott
William and Wilkins: 826 p.
Sallie R, Tredger JM, William R. 1991. Drugs
and the liver. Biopharmaceut. Pharmaceutical
Disposal 12, 251-259.
Schliess F, Görg B, Fischer R, Desjardins P,
Bidmon HJ, Herrmann A, Butterworth RF,
Zilles K, Häussinger D. 2002. Ammonia induces
MK-801- sensitive nitrate and phosphorylation of
protein tyrosine and residues in rat astrocytes. The
FASEB Journal 16(7), 739-741.
Subash S, Subramanian P. 2010. Morin improves
the expression of urea cycle enzymes in
hyperammonemic rats. Journal of Pharmacy
Research 3(6), 2557-2560.
Subash S, Subramanian P. 2012. Protective effect
of morin on lipid peroxidation and lipid profile in
ammonium chloride-induced hyperammonemic rats.
Asian Pacific Journal of Tropical Medicine 1(2), 103-
106.
http://dx.doi.org/10.1016/S2222-1808(12)60025-5
Takeuchi K, Ohuchi T, Harada H. 1995. Irritant
and protective action of urea-unease ammonia in rat
gastric mucosa. Digestive Diseases and Sciences
40(2), 274-281.
Thenmozhi AJ, Subramanian P. 2011.
Antioxidant potential of Momordica charantia in
ammonium chloride- induced hyperammonemic rats.
Evidence-Based Complementary and Alternative
Medicine. Article ID 612023, 7 pages.
http://dx.doi.org/10.1093/ecam/nep227
Thophon S, Kruatrachue M, Upthan ES,
Pokethitiyook P, Sahaphong S, Jarikuan S.
2003. Histopathological alterations of white seabass,
Lates calcarifer, in acute and subchronic cadmium
exposure. Environmental Pollution 121, 307–320.
Stevens A, Lowe J, Scott I, Damjanov I. 2009.
Core pathology, 3 rh ed. Mosby, Elsvier, London: 632
p.
Steven W, Rajiv J, Dejong H. 2009. Interorgan
ammonia trafficking in liver disease. Metabolic Brain
Disease 24, 169-181.
http://dx.doi.org/10.1007/s11011-008-9122-5
Tietz NW. 1995. Clinical guide to laboratory tests,
3 rd ed. W. B. Saunders, Philadelphia: 610-611 p.
Treem WR. 1994. Inherited and acquired
syndromes of hyperammonemia and encephalopathy
in children. Seminars in Liver Disease 14, 236-258.
Ujjwal K, Dey S. 2010. Evaluation of organ
193 Al-Ali et al.

Int. J. Biosci. 2016
function and oxidant/ antioxidant status in goats with
sarcoptic mange. Tropical Animal Health and
Production 2, 1663-1668.
http://dx.doi.org/10.1007/s11250-010-9618-y
Vijayakumar N, Subramanian P. 2010.
Neuroprotective effect of Semecarpus anacardium
against hyperammonemia in rat. Journal of Pharmacy
Research 3, 1564-1568.
http://jpronline.info/article/view/2815/1489
Wills MR, Savory J. 1981. Biochemistry of renal
failure. Annals of Clinical and Laboratory Science
11(4), 292-299.
Williams M. 2006.The Merck index: An
encyclopedia of chemicals, drugs, and biologicals, 14 th
ed., Station, New Jersey: 2564 p.
Wood CM. 1993. Ammonia and urea metabolism
and excretion. In: Evans, D.H. (Ed.). The physiology
of fishes. CRC Press, Boca Raton, Florida: 379-425 p.
194 Al-Ali et al.