الفهرس 
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Abstract
The present work aimed to study the immunomodulatory effect of EPSs isolated from Lactic acid bacteria upon chemically-induced hepatocellular tumors in male rats’ model.
The bacterial exopolysaccharides were extracted, purified and identified. Extracted EPSs were characterized by using FTIR which showed different stretches of bonds at different peaks.
A total number of 50 male Swiss albino rats were randomly assorted into five groups (10 for each) as follows:
group 1 ”Normal Control”: Animals served as control and they received physiological saline daily orally by gastric gavage for eight constitutive weeks.
group 2 ”DEN”: Animals received DEN orally by gastric gavage (diluted in 0.9% normal saline), in a dose of 20 mg/kg b.w. daily five days per week for eight constitutive weeks according to the modified method of Darwish and El-Boghdady (2011).
group 3 ”EPSs”: Animals received EPSs at dose level of 100 mg /kg b.w. daily five days per week (Zahran et al., 2017) orally by gastric gavage for eight constitutive weeks.
group 4 ”EPSs+DEN”(prevention): Animals received DEN orally (20 mg/kg b.w.) daily five days per week for eight weeks, in parallel with administration of EPSs (100 mg EPSs/kg b.w.) daily five days per week orally by gastric cavage.
group 5 ”DEN+EPSs”(treatment): Animals received DEN (20 mg/kg b.w.) daily five days per week for eight constitutive weeks followed by administration of EPSs (100 mg EPSs/kg b.w.) daily five days per week for the next eight weeks.
At the end of the experiment, rats were sacrificed. After an overnight fast from the last doses of EPSs and DEN, the animals were anesthetized using diethyl ether. Blood samples from each rat were obtained via heart puncture and left to coagulate and serum was obtained after centrifuged for biochemical analysis.
Liver function tests as serum alanine aminotransferase (ALT) and serum gamma glutamyl transferase (γ-GT) activities as well as serum total proteins level were determined. Evaluation by determining the most important free radical scavengers of the antioxidant defense system in liver tissue homogenate including reduced glutathione (GSH) content, also marker of lipid peroxidation malondialdehyde (MDA) level was determined. Immunological parameters were determined in the liver tissue homogenate as interleukin 17 (IL-17), interleukin 10 (IL-10) and transforming growth factor beta-1 (TGF-β1) levels were determined by ELISA technique. Molecular investigation was carried out in the liver tissue homogenate as signal transducer and activator of expression level of hepatic transcription 3 protein (STAT3) and expression level of hepatic mitogen activated protein kinase p38 (p38MAPK) were determined by western blot technique.In addation to toll like receptor 2 (TLR-2) gene expression was determined by quantitative real time PCR technique. Histopathological examination of liver tissue was also performed.
The results revealed that, DEN induced a significant increase in serum alanine aminotransferase (ALT) and gamma glutamyl transferase (γ-GT) activities as well as malondialdehyde (MDA), interleukin 17 (IL-17), transforming growth factor beta-1 (TGF-β1), signal transducer and activator of transcription-3 protein (STAT3), mitogen activated protein kinase p38 (p38MAPK) levels in the liver tissue. The gene expression of liver toll like receptor 2 (TLR-2) gene was also increased. While, there was a significant decrease in glutathione (GSH), interleukin 10 (IL-10) contents in the liver tissue and serum total proteins level compared to normal control. However, prevention and treatment with EPSs ameliorated most of the investigated parameters.
The histopathological observations of liver tissues were in agreement with restored biochemical results, revealed that liver tissues with DEN treatment showed significant inflammatory cells infiltration in portal area, while the liver tissues which exposed to EPSs preventive and treatment groups showed significant improvement upon hepatocytes. The hepatopreventive and antiinflammatory effects against HCC liver injury have been directly certified by improved necrotic accumulation, inflammatory infiltration formation in the liver.
Conclusion:
from the aforementioned results, it is concluded that, prevention and treatment of EPSs results suppressive induction of cytotoxic effects on cancer cells. In conclusion, our findings showed that Lactobacillus acidophilus ATCC 4356 EPSs are efficacious control against hepatocellular carcinoma throughout the modulation of key signaling growth factors associated with inflammation via antioxidant mediated anti-inflammatory and anti-proliferative activities.Prevention effect of EPSs gave best reults than treatment effect of EPSs on HCC.