الفهرس 
INTRODUCTIONOnly 14 pages are availabe for public view
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Abstract
Hepatocellular carcinoma is the most common type of primary liver cancer, with
the highest incidence and mortality rate globally. Many risk factors contribute to the
development and progression of HCC, such as chronic HCV, HBS, chronic untreated
inflammation of liver, oxidative stress and fatty liver disease. In Egypt, HCC ranks the
fourth most common cancer and is the second cause of cancer mortality in both sexes and
HCV is the most important risk factor for HCC development in Egypt.
Several protocols are used in the treatment of HCC but they are also associated
with numerous side effects. Many natural products are helpful in the prevention and cotreatment of HCC through many ways as: protecting against liver carcinogens, enhancing
effects of chemotherapeutic drugs, suppression of oxidative stress, chronic inflammation,
and inhibiting tumor cell growth and metastasis.
This study was designed to investigate the phytochemical content of the Egyptian
carob pod aqueous extract (CPAE) and its possible biological activities. Moreover, the
study aimed to investigate the biochemical, Hematological, molecular, and histological
changes in DEN/CCl4 induced HCC in rats. Also, investigating the potential therapeutic
action of CPAE, cisplatin or their combination against DEN/CCl4 induced HCC in rats.
The most important biological activities of CPAE that were measured are
antioxidants, antimicrobial, and antidiabetics. In addition, the phytochemical compounds
were characterized chemically and by using HPLC.
To achieve the aim of investigating the therapeutic action of CPAE, we conducted a
study on 42 male albino Wistar rats (weight range150-170 g and age 2-3 months). Rats
were divided into two groups
A- Control non-HCC group which assigned into 3 groups
1- group 1: Normal control group (n=6): which fed with a normal basal diet
for 36 weeks.
2- group 2: Control treated group with CPAE (n=6): rats of these group were
fed with a normal basal diet for 32 weeks and after that administrated 200
mg/Kg/day CPAE orally via gavage tube for 4 weeks.
3- group 3: Vehicle group (n=6): Which are injected intraperitoneal (IP) with
a single dose 100 mL/kg 0.9 NaCl after two weeks with Olive oil 0.25mL/Kg
for 36 week.B- HCC group which assigned into 4 groups: the animal model of HCC was
induced by using a single IP dose of Diethylnitrosamine (DEN) 100 mg/kg body
weight, two weeks later rats injected IP with 0.25mL/kg carbon tetrachloride
dissolved in olive oil twice for 32 weeks
1- group 1: Control HCC untreated group (n=6)
2- group 2: HCC treated group orally with 200mg/kg of CPAE daily for 4
weeks (n=6).
3- group 3: HCC treated group IP with 0.5 mg/kg of cisplatin as a reference
of chemotherapy drug twice a week for 4 weeks (n=6).
4- group 4: HCC treated group with a combination therapy of CPAE 200
mg/kg daily plus 0.5 mg/kg of cisplatin twice a week for 4 weeks (n=6).
After the end of treatment, all the rat groups were sacrificed to obtain blood for the
assessment of biochemical and hematological investigations. The liver was obtained for
histological investigations, assessment of oxidative stress and antioxidant parameters,
assessment of hepatic gene expression of AMPK, PGC-1α, SIRT-1, TFAM, and iNOS
using RT-PCR, and assessment hepatic protein of pP53, NF-κB, IKK, and SREBP-2
using western blot technique.
The HCC rats showed significant histological alterations. Also, it showed a
significant increment in the percentage of inflammatory cells, necrotic cells, deposition
of fats,collagen fiber deposition, severe hemorrhage, and marked dilation of the blood
vessel. These histological changes were associated with marked elevation in the serum
activities of ALT, AST, and ALP, levels of AFP, bilirubin, urea, and creatinine, and a
marked decline in the AST/ALT ratio, total protein, and albumin. Moreover, the HCC
rats showed significant metabolic alteration which was indicated by the elevation of
fasting blood glucose and triglyceride. The hematological parameters of the HCC rats
showed a significant decline in HB, RBCs, WBCs, and platelets. In addition, the HCC
rats showed a significant upregulation in the hepatic gene of SIRT1, SREBP-2, iNOS,
NF-κB, and IKK. On the other hand, the level of AMPK, PGC-1α, TFAM, and p53 were
significantly downregulated.The treatment of HCC rats with CPAE, cisplatin, and or their combination showed
significant amelioration of all parameters (Biochemical, hematological, histological, and
molecular). For most of the studied parameters, the best effects were observed in the
HCC rats treated with combination therapyfrom the results of the present study we can conclude that:
1- This study revealed that CPAE contained flavonoid, phenolic, alkaloid, amino
acids, and carbohydrates.
2- The HPLC analysis indicated that CPAE contained a high content of gallic acid,
catechin, and protocatechuic acid which make it a good source of antioxidants.
3- CPAE has a significant antioxidant and antihemolytic activity.
4- CPAE has an anti-diabetic effect as it inhibited the digestive enzymes (maltase,
sucrase, lactase, and amylase).
5- CPAE has a strong antimicrobial activity against Staphylococcus aureus,
Escherichia coli, Bacillus subtilis, Candida albicans, and HSV1.
6- The hepatocarcinogenic animal model of DEN/CCl4 was easily established and
mimicked the pathophysiology of HCC in humans.
7- The used model of HCC is associated with impairment of carbohydrate and lipid
metabolism.
8- There is an important crosstalk between AMPK, PGC-1α, and mitochondrial
functions (metabolism, biogenesis, and redox homeostasis). That may put AMPK
and PGC-1α as a cornerstone in treating and preventing the development and
progression of HCC.
9- There is an important crosstalk between NF-κB and iNOS in the progression of
inflammation in HCC rats. Which put NF-κB and iNOS as an important
molecular therapeutic targets for HCC treatment.
10-The treatment of HCC rats with CPAE, cisplatin, or their combination showed
significant correction at nearly all levels of hepatic derangement and tumor
marker AFP.
11-The treatment of HCC rats with CPAE showed significant amelioration of kidney
function impairment while cisplatin showed significant elevation of kidney
function, which indicates a state of nephrotoxicity.
12-The treatment of HCC rats with CPAE, cisplatin, or their combination showed
significant correction of redox status parameters.
13-The treatment of HCC rats with CPAE, cisplatin, or their combination showed
significant correction of metabolic alteration by decreasing the level of blood
glucose, cholesterol, TG, and SREBP-2.14-The treatment of HCC rats with CPAE, cisplatin, or their combination showed a
significant increase in mitochondrial biogenesis as observed by the upregulation
of AMPK, PGC-1α, and TFAM gene expression.
15-The treatment of HCC rats with CPAE, cisplatin, or their combination showed a
significant anti-inflammatory action as observed by the downregulation of iNOS,
NF-κB, and IKK.