2. Detoxifying and antioxidant
effect of ellagic acid
nano particles in rats intoxicated
with sodium nitrites
Amira Ragab EL Barky, Tarek Mostafa Mohamed and Ehab Mostafa
Mohamed Ali.
The Korean Society for Applied Biological Chemistry(KSABC)
3. Introduction
• Sodium nitrite is used to stabilize meat colour, reduce rancidity.
• Nitrites are used in medical field such as vasodilator, bronchodilator,
detoxifying of cyanide poisoning.
• Use of nitrite leads to human exposure to many disease.
• Sodium nitrite is also a carcinogen.
• Ellagic acid found in pomegranate is a powerful anti-oxidant,
antibacterial, antiviral and anti inflammatory compound and can
upregulate tumor suppressor and apoptotic genes.
4. Materials and method
Extraction of ellagic acid
• Ellagic acid was extracted from the pomegranate peel powder.
Synthesis of EA nanoparticles coated with chitosan (EANP@CS)
• EANP@CS was synthesized using sodium triphosphate as a gelatin
agent with some modification.
5. Identification of ellagic acid and EANP Fourier transforms Infrared
spectroscopy(FT-IR)
• To identify the functional groups of the active compound, an infrared
spectroscopy was performed.
7. X ray diffraction
• X ray diffraction patterns for EA and EANP@CS were recorded.
8.
9. Experimental animals
50 adult white male albino wister rats 3-4 months (180-200g)
Normal group
Sodium
nitrite group
Control rats – no drugs
Receiving EANP@CS
(20 mg/kg)
Untreated nitrite group
(2g/L nitrite for 8 weeks)
Sodium nitrite + EANP@CS
(Treated with EANP@CS for
4 weeks)
EANP@CS treated 4 days/
week after stopping
sodium nitrite
10. • By the end of 8 weeks rats were sacrificed
• Serum was used for evaluation for various biochemical parameters.
• The liver kidney and spleen was separated and used in two parts one
part washed and homogenized. The second part fixed and stained
then used in pathological study.
11. Statistical analysis
• Excel 2013, Origin 6 and SPSS 19 were used to perform statistical
analysis.
• One-way ANOVA with post hoc Tukey’s tests were carried out.
• Ryan-Einot-Gabriel-Welsch F test was performed to separate means by
using SPSS program.
12. Determination of Hemoglobin; urea level, ALT and AST activities
• Serum urea – biomarker for kidney disease
• ALT and AST – for liver disease
• Liver and kidney protein were measured
13.
14. Determination of L-MDA; GSH, total protein thiol(PTH) and non protein
thiol (NPTH) levels; Gpx; GST and catalase activities
• MDA level calculated according to Draper and Hadley method.
• GSH measured in liver and kidney
• Total protein thiol (PTH) and non protein thiol (NPTH) Performed in
the liver and kidney.
• Gpx activity was determined in liver and kidney by Ellman method
15.
16.
17. Determination of serum, liver and kidney NO, liver NOS, liver and
kidney arginase activities and DNA fragmentation
• Total nitrite was determined by Griess’s reagents
• The level of nitric oxide(NO) (nitrite and nitrate) was determined in
blood serum and liver homogeneity
• DNA fragmentation was performed according to Gercel-Taylor
method.
18.
19.
20. Preparation of samples for histopathology
• Parts of liver, kidneys and spleen were processed for histopathological
studies
21.
22.
23.
24. Conclusion
• Ellagic acid contains powerful antioxidants
• EANP@CS can be used as potent drug against harmful activities of
carcinogen like sodium nitrites.
