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Biochemical and histopathological response of Swiss albino mice treated with uranyl nitrate
Sangeetha Vijayan P
Junior Research Fellow
Yenepoya Research Centre
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Background
• Human exposure to heavy metals such as uranium is of great concern to
our society.
• The wide spread uses of uranium for instance in the civilian and military
applications, in nuclear power, counterweights in aircrafts and penetrators
in shrapnel increase the release of it to the environment.
• Uranium dispersion causes contamination of ground water as well as food
chain and poses treat to the health of general population.
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Background
• Animals as well as human studies have shown the chemical toxicity of uranium
• Toxicity of uranium varies with chemical form. More soluble uranium
compounds have highest toxicity.
• Inhalation, ingestion or skin contact is the main route of exposure to uranium
and leads to respiratory diseases, kidney failure, reproductive diseases etc.
• Kidney is the most sensitive organ for uranium toxicity. (Gilman et al., 1998)
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Toxic effects of Uranium of uranium
• Nephrotoxicity
• Hepatotoxicity
• Developmental toxicity
• Reproductive toxicity
• Bone toxicity
( Brugge et al., 2011)4
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Effect of uranium on nephron
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AIM
• To determine the effect of Uranyl nitrate on mice kidney at different time
intervals with respect to biochemical and histopathological changes
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Materials and Methods
• Animal model: Swiss albino mice
• 8–10 weeks of age, weighing 25 ± 5g
Study groups
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Treatment groups Treatment Uranyl nitrate
Dose (mg/kg b.wt)Control (Group1) 0.9% saline
Day 1 (Group II)
UN treated
1 2 4
Day 3 (Group III )
UN treated
2 4
Day 5 (Group IV)
UN treated
2 4
Day 14 (Group V )
UN treated
2 4
Day 28 (Group V1 ) UN treated 2 4
Each study group contains 5 animals (males )
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Material and methods….
Biochemical analysis
• Biochemical parameters such serum urea, creatinine and blood urea
nitrogen was estimated.
• Biological diagnostic kits (Aggappe Diagnostic Ltd, India) were used and
biochemical analyses were carried out using a biochemical analyzer
(MispaPlus, India) as per the manufacturer’s protocols.
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Materials and Methods…
Histopathological analysis
Tissue processing and staining
Kidneys were collected immediately after sacrifice of the mice and fixed in
10% neutral buffered formalin over night
▼
Fixed kidney specimens were dehydrated in graded ethanol, cleared in xylene
and infiltrated with paraffin wax using an automatic tissue processor
▼
Processed specimens were embedded in paraffin wax using metallic blocks
▼
5 µm thick sections of kidneys were taken in a rotary microtome
▼
Sections were stained with Hematoxylin and Eosin counterstains (H-E) and
observed at ×400 magnification under a light microscope (Motic BA210, Hong
Kong).)) 9
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RESULTS
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Figure 1: Difference in body weight of Swiss albino mice treated with 2mg/kg bd wt of uranyl nitrate at different time intervals
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0 1 3 5 14 285
10
15
20
25
30
35
initial wtFinal wt
Days
Bod
y w
eigh
t (g
)
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Figure 2: Difference in body weight of Swiss albino mice treated with 4mg/kg bd wt of uranyl nitrate at different time intervals
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0 1 3 5 14 285
10
15
20
25
30
35
Initial bdwtFinal bdwt
Days
Bod
y w
eigh
t (g
)
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Control UN0
10
20
30
40
50
0
0.05
0.1
0.15
0.2
0.25Urea BUN Creatinine
Treatment groups
Ure
a an
d B
UN
(m
g/dL
)
Cre
atin
ine
(mg/
dL)
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Fig 3: Serum biochemical parameters (Urea, Creatinine and BUN) of 1 mg/kg bd wt uranyl nitrate treated groups in comparison to control. Statistical difference was not observed between the groups (p > 0.05). Vertical bars represent mean ± S.D., n = 5
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14
Control 1 3 5 14 280
0.4
0.8
1.2
1.6
2mg 4mg
Days
Ser
um c
reat
inin
e (m
g/dL
)
Fig 4: Serum creatinine levels of uranyl nitrate treated groups (2 mg/kg bdwt and 4 mg/kg bdwt ) in comparison to control. Statistical difference was observed between the groups (p < 0.05). Vertical bars represent mean ± S.D., n = 5
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.
control 1 3 5 14 280
100
200
3002mg 4mg
Days
Seru
m u
rea
mg/
dL
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Fig 5: Serum urea levels of uranyl nitrate treated groups (2 mg/kg bdwt and 4 mg/kg bdwt ) in comparison to control. Statistical difference was observed between the groups (p < 0.05). Vertical bars represent mean ± S.D., n = 5
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Figure 6: Histopathology of mice kidney treated with uranyl nitrate
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1mg/kg 1 day
2mg/kg 1 day 4mg/kg 1 day
Cortical region of kidney(H-E 400X)
Control
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Figure 7: Histopathology of mice kidney treated with Uranyl nitrate
17Cortex region of kidney. Arrows indicates the casts in tubular lumen
2mg,3 days4mg 3 days
4mg 5 days
2mg 3 days
2mg 5 days2mg 5 days
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Figure 8: Histopathology of mice kidney treated with Uranyl nitrate
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2mg 14 days 4mg 14 days
4mg 28 days
Cortex region of kidney. Arrow indicates damage in proximal convoluted tubule
2mg 28 days2mg 28 days
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Conclusions
• Pathological alternations in kidney cells and elevated levels of serum creatinine and
urea was shown at a dose of 4 mg/kg (ip)after 3 and 5 days of uranyl nitrate
• Completed recovery was showed after 14 days in mice treated with 2 mg/kg bdwt of
uranyl nitrate
• Mice treated with 4mg/kg Uranyl nitrate showed almost recovery after 28 days
• A dose of 4 mg/kg after 3 days to induce maximum nephrotoxicity in mice with
respect to both biochemical and histopathological analysis.
• Dose and time depended increase in toxicity and recovery was showed in animals
treated with uranyl nitrate 19
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References…
1. ATSDR 2013. Toxicological profile for Uranium.U.S. Department of Health and Human Services. Public Health Service.
Agency for Toxic Substances and Disease Registry. Atlanta, GA.
2. Brugge D and Buchner V. Health effects of uranium: new research findings. Rev Environ Health.2011;26(4):231–249.
3. Basile DP, Anderson MD, Sutton TA. Pathophysiology of acute kidney. Comprehensive Physiology. 2012;2(2):1303-1353
4. Hodge HC, Stannard JN, Hursh JB. A review of the many aspects of uranium toxicity from animal studies. New York:
Springer-Verlag; 1973; 165-96.
5. Gilman AP, Villeneuve DC, Secours VE, Yagminas AP, Tracy BL, Quinn JM, Valli VE, Willes RJ, Moss MA. Uranyl
nitrate: 28-day and 91-day toxicity studies in the Sprague-Dawley rat. Toxicol Sci.1998;41:117-128
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