Efficiency of cape gooseberry in attenuating some biochemical disorders and oxidative stress...

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62 [email protected]

Efficiency of cape gooseberry in attenuating some

biochemical disorders and oxidative stress associated

with hepatocellular carcinoma

Hanaa M. Serag*, Hanaa A. Hassan, Makwan S. Qadir

Physiology Division, Zoology Department, Faculty of Science, Mansoura University, Mansoura, Egypt

* E-mail: [email protected]

Abstract

Hepatocellular carcinoma (HCC), also called malignant hepatoma is the most common type of liver

cancer. The aim of the present study is to elucidate the role of naturoceutical agent such as cape

gooseberry (CG) (Physalis peruviana) as a chemo-sensitizer for adriamycine (ADR) treatment the

hepatocellular carcinoma rats model. The present data recorded that HCC rats has a significant

increase in serum and hepatic lipids profile (TL, TC, TG, LDL-C and VLDL-C) accompanied with a

significant decrease in HDL-C level. Also, total protein (TP) and zinc (Zn) and copper (Cu) levels

were decreased. Moreover, the data illustrated a marked increase in total bilirubin, AFP level as well as

enzymes (AST, ALT, ALP, γ-GT) activity, however hepatic ALT and AST activity was significantly

decreased in HCC rats. In addition HCC rats has a significant increase in hepatic oxidative stress

markers (MDA & NO) and free radicals enzymes (AO & XO), but hepatic antioxidant status including

GSH, TAC, SOD, catalase, and GSH-PX was significantly reduced in HCC rats. On the other hand

HCC rats received ADR or CG showed an improvement in all the above parameters through the

occurred amelioration of the obtained alterations of lipids profile, liver function enzymes, oxidative

stress and antioxidant defense system. Furthermore the data evidenced that CG is more effective than

ADR and it act as a chemo-sensitizer for ADR treatment of hepatocellular carcinoma.

Keywords: Hepatocellular carcinoma, Cape gooseberry (Physalis peruviana), Liver functions,

Oxidative stress, Antioxidant defense mechanism

List of abbreviations:

TL, Total lipids; TC, Total cholesterol; TG, Triglyceride; HDL-C, High density lipoprotein- cholesterol; LDL-C, Low density

lipoprotein- cholesterol; VLDL-C, Very Low density lipoprotein- cholesterol; AST, Aspartate aminotransferase; ALT, Alanine

aminotransferase; ALP, Alkaline phosphatase; γ-GT, Gama-Glutamyle Ttransferase; AFP, Alpha –Fetoprotein; MDA,

Malondialdehyde; NO, Nitric oxide; AO, Aldehyde oxidase; XO, Xanthine oxidase; SOD, Superoxide dismutase; GSH,

Glutathione; TAC, Total antioxidant ; capacity; GSH PX, Glutathione peroxidase; CG, Cape gooseberry

Vol 22, No. 11;Nov 2015


1. Introduction

Hepatocellular carcinoma (HCC) represents the most common primary malignancy of the liver.

According to epidemiological survey, the prevalence of HCC ranks the sixth among all cancers. HCC

accounts for approximately 700,000 cancer-related deaths per year, which ranks thethird in global

cancer statistics (Jemal et al., 2011). Most cases of HCC are secondary to either a viral hepatitis

infection (hepatitis B or C) or cirrhosis (alcoholism being the most common cause of hepatic cirrhosis).

According to recent reports, the incidence of HCC has increased sharply in the last decade, especially

in Egypt, where there has been a doubling of the incidence rate during the last 10 years. This sharp rise

has been attributed to several factors such as biological including hepatitis B and C virus infection,

endemic infections in the community as schistosomiasis as well as environmental factors including

aflatoxin (Arzumanyan et al., 2013). Other factors such as cigarette smoking, occupational exposure to

chemicals as pesticides may also contribute to the etiology or progression of the disease (Anwar et al.,

2008). Liver carcinogenesis may also develop through the progressive accumulation of different

mutations (genetic) and/or gene products (protein), which eventually lead to malignant transformation

(MacPhee, 1998; Benzie, 2003 and Seufi et al., 2009).

Although, the diagnosis and treatment of HCC have significantly been improved in recent years, the

prognosis remains poor (El-Serag, 2011). Systemic chemotherapies have been evaluated for many

years. Anticancer drugs are widely used against variety of human cancer. However, while they generate

acceptable outcome in chemotherapy of some cancers, they also exhibit severe toxicity and undesirable

side effects (Minami et al., 2010). Adriamycin (an antibiotic isolated from Streptomyces peucetiusvar)

is considered one antitumor drug commonly used as single and in combination with other

chemotherapeutic agents, for treatment of wide range of human malignancies (Stewart and Ratain,

2001), such as multiple myeloma (Alberts and Salmon, 1975), osteogenic sarcoma (Wang et al., 1971),

lymphocytic leukemia (DiMarco et al., 1969), HCC patients (Carr and Nagalla, 2010) and stomach

cancer (Ogawa, 1985). Due to the increasing awareness of the side effects of conventional medicine,

the use of natural products as an alternative to conventional treatment in the healing and treatment of

various diseases, including cancer, has been increasing in the last few decades (Kaefer and Milner,

2008). Several herbal drugs have been evaluated for their potential to protect the liver against

hepatotoxicity (Ramakrishnan et al., 2006).The medicinal value of plants have assumed an important

dimension in the past few decades owing largely due to the discovery as a rich source of antioxidants

that combat oxidative stress through their redox active secondary metabolites (Hassan et al., 2014).

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Among these plants, cape gooseberry (Physalis peruviana), cape gooseberry (CG) is an annual,

herbaceous plant which belongs to Solanaceae family, its fruit, also known as golden berry, ground

cherry and in Egypt called harankash (Figure 1). The fruit of cape goosberry is highly nutritious,

having high levels of essential minerals, magnesium, calcium, potassium, sodium and phosphorus

which are classified as macronutrients, while the Iron and Zinc are considered as micronutrients

(Szefer and Nriagu., 2007);vitamins A, B and C. The main active components of vitamin A in fruits are

α-carotene, β-carotene and β cryptoxanthin. Moreover it contains biologically active components e.g.

physalins, withanolides, phytosterolsand polyunsaturated fatty acids e.g. linoleic acid andoleic acid. Its

bioactive compounds have nutritional value, medicinal properties as well as strong antioxidant

property that can prevent peroxidative damage of liver microsomes and hepatocytes (Dinan et al., 1997

and Wang et al., 1999). Cape gooseberry fruit and other aerial parts are used in the treatment of

intestinal and digestive problems and used as mutagenic, anticoagulant, antispasmodic, antileucemis

agents (Shariff et al., 2006 and Helvaci et al., 2010). Epidemiological studies from other parts of the

world indicate that increased consumption of fruits and vegetables are associated with lower risk of

cancer, leukemia, hepatitis and various chronic degenerative diseases (Reddy et al., 2010; Zhang et al.,

2013).

So, the aim of the present study is to elucidate the potential role of cape gooseberry (Physalis

peruviana) as a chemo-sensitizer for adriamycine treatement of experimental hepatocellular carcinoma

rats model.

2. Materials and methods

2.1. Animals

The healthy male albino rats (Rattus rattus), 8 weeks old, weighing 150-170g were used in this study.

Rats housed in a stainless steel cages in a windowless room with automatically regulated temperature

(22-25°C). They were kept under good ventilation under a photoperiod of 12 h light: 12 h darkness

schedule with lights-on from 06.00 am to 18.00 am. They were received a standard laboratory diet

composed of 60% ground corn meal, 15% ground beans, 10% bran, 10% corn oil, 3% casein, 1%

mineral mixture and 1% vitamins mixture, purchased from Meladco Feed Company (Aubor City,

Cairo, Egypt) and supplied with water ad libitum throughout the experimental period. Animals

received human care and the present study complies with the instruction’s guidelines. The local

committee approved the design of the experiments, and the protocol conforms to the guidelines of the

National Institutes of Health (NIH).

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2.2. Chemicals and drugs

Diethylnitrosamine (DENA) and CCl4were purchased from Sigma Chemical Co. (St. Louis, MO,

USA). DENA was freshly dissolved in sterile 0.9 % saline and given to rats at a single dose of 200

mg/kg body weight (Sarkar et al., 1997). CCL4 was given to rats at a dose of 3ml/kg/ body

weight/week (Subramanian et al., 2007; Singha et al., 2009). Adriamycin (ADR) was purchased from

Tarshouby Pharmacy, Mansoura , Egypt and subjected at a dose of 2 mg/kg body weight once per

week (Sakr et al., 2011).

2.3. Preparation of cape gooseberry juice

Cape gooseberry (Physalis peruviana) was purchased from local markets, Mansoura, Egypt. Fruits

were first washed thoroughly to remove impurities. After washing the fruits, they were cut into small

pieces, freshly prepared juice in the blender (500g Cg juice up to 500ml distled water, where each 1 ml

juice contains 1g Cg). The juice of cape gooseberry (Cg) (1 ml/kg body weight) was shaken well just

before oral administration by gavage (Arun and Asha, 2007).

2.4. Experimental design

After two weeks of acclimatization, the rats were divided into five groups 6 rats each group. Group

1(Control): the rats of this group did not receive any treatments. Group 2 (HCC): the rats treated with a

single intraperitoneal injection of freshly DENA (200 mg/kg body weight). Two weeks later, they

received a subcutaneous injection of CCl4 (3 ml/kg/week) for 10 weeks to promote the carcinogenic

effect of DENA. Group 3: (HCC+ADR): HCC rats were injected intraperitonealy with ADR at a dose

of 2 mg/kg body weight, once per week for 10 weeks. Group 4: (HCC+CG): HCC rats were daily

administered cape gooseberry (Cg) juice at a dose of 1 ml/kg/bw. Group 5: (HCC+CG +ADR): HCC

rats were injected intraperitonealy with ADR at a dose level of 2 mg/kg body weight, once per week

and were administered with cape gooseberry (CG) at a dose of 1 ml/kg/bw.

2.5. Blood and liver sampling

At the end of the experimental period (12 week), fasted overnight rats were sacrificed by cervical

dislocation and blood samples were collected into clean centrifuge tube and subjected to centrifugation

at 860 Xg for 20 min at 4OC. The separated sera were frozen at – 20

OC for further analysis. The rats

were dissected and the livers were immediately excised, rinsed with ice-cold saline, blotted dry and

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accurately weighed. They were minced and homogenized in ice cold buffered saline (10% w/v). The

homogenates were centrifuged at 860 Xg for 10 min at 4 °C. Finally the supernatants were subjected to

the biochemical analysis.

2.6. Biochemical analysis

Total lipids level was estimated according to the technique of Zollner and Kirsch (1962). Total

cholesterol and triglycerides levels were estimated using kits (Biodiagnostic for laboratory services,

Egypt.) according to Allain et al. (1974) and Fassati and prencipe (1982), respectively. High-density

lipoprotein cholesterol (HDL-C) was determined by using enzymatic colorimetric method of

Lopez,(1977). Low-density lipoprotein choleseterol (LDL-C) was calculated according to the

following equation: LDL-C = TC– HDL-C – TG/5, as described by Friedewald et al. (1972). Very

low-density lipoprotein (VLDL-C) was calculated according to the following equation:

VLDL-C=TG/5, as described by Satheesh and Pari (2008). The levels of total protein , total bilirubin,

Zn and Cu were estimated using instructions of kits supplied by Bio-diagnostic Co., Mansoura, Egypt

as described by Henry (1964); Jendrassik and Grof (1938); Hayakawa and Jap (1961) and Ventura and

king, (1951), respectively.

AFP level was estimated by immunoenzymatic colorimetric method according to Acosta et al. (1983).

Aspartate transaminase (AST) alanine transaminase activity (ALT) and Alkaline phosphatase (ALP)

activity was measured using colorimetric Kits purchased by ABC diagnostic kit, Cairo, Egypt as

described by Reitman and Frankel (1957) as well as Belfield and Goldberg (1971),respectively.

Gamma-glutamyl transpeptidase (γ-GT) activity was estimated according to the method of Szasz (1969)

using γ-GT-kit purchased from SGM Diagnostic division, Roma, Italy. The content of

malondialdehyde (MDA) was estimated according to the colorimetric method of Ohkawa et al. (1982).

Nitric oxide (NO) level was determined by colorimetric method of Montgomery and Dymock (1961).

Hepatic aldehyde oxidase (AO) xanthine oxidase (XO) activity was assayed as described by the

method of Johnson et al. (1984) and Stripe and Della Corte (1969), respectively. Glutathione content

(GSH) was adopted by Beutler et al. (1963).Total antioxidant capacity was determined according to

the technique of Koracevic et al. (2001) using colorimetric Biodiagnostic Com. Kit (29 Tahreer St.,

Dokki,Giza, Egypt). Superoxide dismutase (SOD) activity was assayed by the procedure of Nishikimi

et al. (1972) .Catalase activity was determined by the colorimetric method of Aebi (1984). Glutathione

peroxidase (GPX) activity was measured by a colorimetric method of Paglia and Valentine (1967).

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2.7. Statistical analysis

Results were expressed as means ± SE. Statistical significance was calculated using one- way analysis

of variance (ANOVA) followed by Duncan′s multiple range tests (Waller and Duncan, 1969). All the

statistical analysis was carried out with the use of SPSS 12.00 software. Differences were considered

significant at P ≤0.05.

3. Results

The present data showed that HCC rats have a significant increase in serum and hepatic lipid profiles

(TL, TC, TG, LDL-C and VLDL-C) levels but only serum HDL-C level was significantly decreased if

comparing to control rats group (table 1). Also, total protein content as well as Zn and Cu level was

significantly decreased in HCC rats group compared to the control rats. Additionally, an increase was

recorded in serum total bilirubin, AFP, AST, ALT, ALP, γ-GT, hepatic ALP and γ-GT in HCC rats if

comparing to control rats group. However hepatic ALT and AST was significantly decreased in HCC

rats comparing to control rats group (table 2). Moreover, the present data indicated that HCC rats have

a significant increase in hepatic MDA and NO level as well as AO and XO enzymes activity associated

with a marked depletion in hepatic GSH content and TAC as well as SOD, catalase, and GSH-PX

enzymes activity if comparing to control rats group (table 3). Moreover, the data presented in table 1, 2,

3 illustrated that ADR treatment showed little amelioration of the obtained alterations of the liver

indicative biomarkers. On the other hand rats that received CG in single or in combination with ADR

showed marked improvement in all the above parameters as evidenced by upgrade the adverse changes

of lipid profiles, liver function enzymes, oxidative stress and antioxidant defense system.

4. Discussion

Hepatocellular carcinoma is the most frequent primary malignancy of the liver, and it accounts for as

many as one million deaths worldwide in a year. In some parts of the world it is the most common

form of internal malignancy and the most common cause of death from cancer (Jemal et al., 2005).The

main obstacles for systemic chemotherapy in HCC include chemoresistance of HCC cells, and

intolerance to the cytotoxic drugs in cirrhotic patients (Wang et al., 2010). Adriamycin is the most used

single effective agent in HCC (Lai et al., 1988). Traditional medicine, especially the herbal medicine

plays a vital role in the management of various liver disorders. Epidemiological studies have shown

that fruits, vegetables, beverages, spices, tea and medicinal herbs rich in antioxidants and other

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micronutrients protect against diverse forms of chemically-induced hepatic damage, carcinogenesis,

mutagenesis, DNA-damage and lipid peroxidation (Wattenberg, 1990 ; Hassan and Abdel-Aziz, 2010).

Liver is a major organ, regulates many important metabolic functions, and any injury causes distortion

of these metabolic functions. The liver is responsible for the metabolism of drugs and toxic chemicals,

and therefore is the primary target organ for nearly all toxic chemicals (Wolf, 1999). Most

apolipoproteins endogeneous lipids, and lipoproteins are synthesized in the liver, and depend on the

integrity of the cellular functions of this organ. Under normal physiological conditions, the liver

ensures homeostasis of lipid and lipoprotein metabolism. Hepatocellular carcinoma impairs this

process leading to alterations in the lipid and lipoprotein patterns (Jiang et al., 2006). A marker is

synthesized by the tumor and released into the circulation, but it also may be produced by normal

tissues in response to an invasion by cancer cells (Thangaraju et al., 1998). Alteration in lipid profile

manifested by a significant increase in total lipid (TL), triglycerides (TG), total cholesterol (TC), and

low density lipoprotein cholesterol (LDL-C), and a significant decrease in high density lipoprotein

cholesterol (HDL-C), was observed in HCC rats. The present increase of LDL-C, VLDL-C and

decrease of HDL-C in HCC rats model may be due to DENA induced abnormal lipid synthesis or

defective degradation of lipids are implicated in the pathological condition like cancer. Peroxidation of

lipids in biomembranes and tissues causes the leakage of these lipids into circulation and consequently

leads to hyperlipidemia. Hyperlipidemia has been shown to increase the risk of metastasis in several

cancers (Sako et al., 2004). Hepatoma is usually associated with hyperlipidemia as well as a notable

decrease in the high-density lipoprotein (HDL) fraction and an enormous increase in the VLDL and

LDL fractions (Kawasaki et al., 2004). Furthermore, the major metabolic vital organ, liver, plays a

key role in cholesterol metabolism in mammals. Reports reveal hypercholestrolemia associated with

hepatomas. The LDL cholesterol is more susceptible to oxidation in various pathologic conditions

resulting in higher lipid peroxidation (LPO) during oxidative stress (Regnstrom et al., 1992). The

occurred DENA-induced hepatic dysfunction may be attributed to its interaction with mitochondria

electron transport and the energy required for protein synthesis by the hepatocytes depending partially

on the catabolic action of glucocorticoids, which could explain the present decreased total protein

content (Dakshayani et al., 2005). Liver dysfunction is reflected by elevation in serum bilirubin

concentration. Bilirubin is a metabolic breakdown product of hem derived from senescent RBCs and

the increase in its level can be indicative to liver dysfunction (Olubunmi et al. 2011). Thus, the present

alterations in the level of TP and bilirubin may indicate liver dysfunction, which may in turn contribute

to metabolic alterations. Also, the viewed data showed increased AFP in HCC rats. This finding

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confirmed by previous study that suggested an elevated serum level of AFP in the adult animals which

are exposed to hepatocarcinogens (Sell and Beck,1978). It was reported that elevated serum protein

can be observed due to rats exposed to hepatocarcinogens and are frequently associated with HCC. Its

serum concentration can be used to confirm hepatocarcinoma and for the diagnosis of tumor response

to therapy. More than 90% of patients with hepatic cancer have increased serum AFP levels (Maideen

et al., 2012). AFP is a glycoprotein which is normally produced by the fetal liver, yolk sac, and the

gastrointestinal tract. AFP is the most commonly used tumor marker for HCC in clinical practice. It is

easily obtainable and relatively inexpensive (Bruix and Sherman.,2005). Although it is most commonly

elevated in HCC, elevations in serum AFP can be seen in various malignancies including testicular,

bile duct, pancreatic, stomach and colon cancer. Elevated AFP can also seen with non-malignant

conditions including hepatitis and cirrhosis (Di Bisceglie et al., 2005).

Moreover, the observed elevation of liver enzymes activity including AST, ALT, ALP and γ-GT in

HCC rats was supported by Shaarawy et al, (2009). These findings may be due to DENA induced

hepatic damage, hepatic dysfunction and subsequent leakage of these enzymes from the neoplastic cell

into circulation (Dakshayani et al., 2005) or may be due to the release of enzymes from normal tissue

by tumor or may be due to possible effect of tumor on remote tissue leading to the loss of its enzyme

and release into the blood (Schwartz and Bodansky, 1965). Also Rocchi et al., (1997) reported that

there is an increase in the levels of these transaminases activity in serum of HCC patients. In

concurrence with the above findings an elevated serum aminotransferase activity was observed in

animals bearing HCC. ALT, which is mainly produced in the hepatocytes, is more specific for liver

injury (Thomson, 1998). It has been reported that ALT is generally increased in situations where there

is damage to the liver cell membrane (Schumann et al 2002). Thus, when the liver is injured, the levels

of ALT in plasma usually rise (Patel et al 1994). Alkaline phosphatase (ALP) is used as a specific

tumor marker during diagnosis in the early detection of cancer (Kobayashi and Kawakubo 1994). It is

involved in transport of metabolites across cell membrane, protein synthesis, secretory activities and

glycogen metabolism. It is a membrane bound enzyme and its alteration is likely to affect the

membrane permeability and produce derangement in the transport of metabolites (Patel et al., 1994).

The present increased of ALP in HCC rats may be due to the disturbance in secretory activity or due to

altered gene expression under these conditions. The development of a tumor results in tissue damage

that lead to the release of ALP into circulation, these result confirmed by Iqbal et al. (2004). Elevation

of alkaline phosphatase is one of the signs, suggesting space-occupying lesions in the liver (Iqbal et al.,

2004). The rise in the activity of ALP in cancer bearing animals may be due to the disturbance in the

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secretory activity or in transport of metabolites, or may be due to altered synthesis of certain enzymes

in these conditions. Also γ-GT t is over expressed in tumor cells (Bailey et al 2001). It is a

membrane-bound enzyme that exhibits a tissue-specific expression and is influenced by various

physiological and pathological conditions, including fetal liver development and hepatic

carcinogenesis (Yao et al., 2004; Tang et al., 1999). γ-GT has been noted to be useful as a specific

HCC marker, and its sensitivity has been reported to be 74% in detecting any size of HCC and 43.8%

in detecting small HCC (Cui et al., 2004). It is involved in the transport of amino acids and peptides

into cells (Hanigan and Pitot, 1985).γ-GT has been shown to play an important role in the metabolism

of foreign substances, and during cell growth and differentiation (Thusu et al., 1991). The observed

increased of γ-GT in HCC rat may be due γ-GT was strikingly activated during the course of

hepatocarcinogenesis induced by several hepatocarcinogens in animals, these result confirmed by Fiala

and Fiala (1973) who suggested that chemical carcinogens may initiate some systematic effects that

induce γ-GT synthesis (Vanisree and Shyamaladevi, 1998; Farombi et al., 2009).

Other related studies indicated that HCC is associated with oxidative stress, as reflected by increased

lipid peroxides with decreases in the antioxidants. Oxidative stress is known to be associated with

increased lipid peroxidation and reduced antioxidant activity (Manimaran and Rajneesh, 2009). An

elevation of both MDA and No level in HCC rats may be due to oxidative conversion of cellular

poly-unsaturated fatty acids to toxic products known as malondialdhyde (MDA) or lipid peroxides

(Dewa et al., 2009). MDA owing to its cytotoxicity and inhibitory action on cellular protective

enzymes is suggested to act as a tumor promoter and a carcinogenic agent (Manimaran and Rajneesh,

2009). MDA is a major end product of lipid peroxidation which can crosslink with DNA and other

protein molecules, thereby it promotes tumoriogenesis (Luczaj and Skrzydlewska, 2003(. In the

present investigation, an increase in MDA formation was presumably associated with increased ROS,

consistent with the observation that these free radicals reduce the activity of hepatic SOD. These result

confirmed by Robak and Glyglewsi (1988). In addition, the occurred increased of NO in HCC rats may

be due to hepatocytes themselves in response to tissue damage and inflammation induced by various

xenobiotics including CCl4. In addition, its role in oxidative stress cannot be neglected, since high

levels of NO have been associated with oxidative injury via lipid peroxide. (Breikaa et al., 2013). NO

plays crucial roles in inflammation and liver injury (Leung et al., 2011). It is produced in large

quantities by Kupffer cells, endothelial cells, and the (Breikaa et al., 2013). NO is a potent cellular

signal, used in a variety of regulatory physiological pathways. However, increased generation of NO is

considered cytotoxicand can lead to tissue damage (Cooper and Magwere, 2008).

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The observed oxidative stress in HCC associated with the recorded elevation of free radicals enzymes

such as AO and XO. The changes in the activity of aldehydes produced by lipid peroxidation have also

been reported in a variety of tumor cells is a metabolizing enzyme, located in the cytosolic

compartment of tissues in many organisms. AO catalyzes the oxidation of aldehydes into carboxylic

acid, and in addition, catalyzes the hydrozylation of some heterocycles (Gordon et al., 1940). Also XO

produce oxidative stress by generating ROS. XO is the most important cellular source of enzymatic

radicals, XO leads to the formation of oxygen radicals and hydrogen peroxide during the 2 steps of

hypoxanthine and xanthine utilization these enzymes catalyze. The oxidation of hypoxanthine

to xanthine and can further catalyze the oxidation of xanthine to uric acid. These enzymes play an

important role in the catabolism of purines in some species, including humans. Xanthine oxidase is an

enzyme involved in several pathways. Some recent studies had noticed that xanthine oxydase

expression is augmented in milk and lower in colostrums; this fact can be involved in the transition

from colostrum to milk production (Hille, 2005). Free radicals react with body tissues and generate

lipid peroxidation, DNA lesions and enzyme inactivation, thus leading to the alteration and impairment

of function of all cellular components leading to apoptosis (Ciriolo, 2005). In the present study

changes in the activity of catalase (CAT), Glutathione peroxidase (GPx), and Superoxide dismutase

(SOD) glutathione reductase (GRD) were investigated. Antioxidants are substances that either directly

or indirectly protects cells against adverse effects of xenobiotics, drugs, carcinogens and toxic radical

reactions (Sen, 1995). The observed decrease in SOD activity suggests inactivation of the enzyme

possibly due to increased superoxide radical production or an inhibition by the H2O2 as a result of

corresponding decrease in the activity of catalase which selectively degrades H2O2 (El shahat, 2013).

The occurred decreased GSH, SOD and CAT in HCC rat groups may be due to accumulation of lipid

peroxidation that emphasized to increase during carcinogenesis with accompanying reduction in

activity of SOD, CAT and depletion of GSH content, suggesting induction of oxidative stress. SOD is

considered the first line of defense against deleterious effects of oxygen free radicals in the cells by

catalyzing dismutation of superoxide radicals (O2¯) to H2O2 and molecular oxygen. CAT is

responsible for detoxification of H2O2, which is an effective inhibitor of SOD (De Duve and

Baudhhuin, 1996). Kregel and Zhang, (2007) attributed the significant decrease in the activity of SOD

and CAT might be due to the excess of ROS, which interacts with the enzyme molecules causing their

denaturation and partial inactivation. The reduction in the activity of CAT may be due to reflect

inability of tissues to eliminate H2O2. SOD enzyme requires zinc and copper for its antioxidant role

(Roughead et al., 1999). The significant changes in total SOD (MnSOD and CuZnSOD) can be

supported by the present deficiency of serum level of Zn and Cu (being essential for regulating cellular

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redox state) which may be related to the reduction of SOD activity in liver of HCC rats exerting

mitochondrial impairments and dysfunction confirming the above explanations. Zinc is thought to have

its own antioxidant property. It thus induces endogenous antioxidants, such as the metallothionines

(DiSilvestro, 2000). Zinc salts have also been reported to exert radical scavenging properties in vitro

(Bagchi et al., 1997). It seems possible that the increased mean serum Zn levels found with the

supplemental intakes of oats may thus improve the antioxidant status by possibly synthesizing zinc-

containing proteins (Bagchi et al., 1997). Moreover, catalase protects SOD against inactivation by

H2O2, while SOD protects catalase against inhibition by (O2¯). Thus, the balance of this enzyme

system may be essentialto get rid of ROS generated in the tissues. In this concern, GSH represents an

important defense mechanism in protecting cells against ROS (Linder, 1995). The decreased of GSH

in HCC rat may be due to the diminished activity of glutathione reductase (GR) (Pulpanova et al.,

1982).The decrease in GSH content in circulation has been reported in malignant states which may

contribute to increased susceptibility to lipid peroxidation (Pasupathi et al., 2009). Synthesized GSH

can be translocated to enter blood circulation and to be taken by organs possessing γ-GT that helps to

metabolize GSH. It has shown that reduction in activity of γ-GT leads to inability to maintain the

constituent amino acids of GSH, resulting in its loss (Griffith and Meister, 1980). The occurred

decreased GPx in HCC rat may be due to of GPx was owed by an enhanced free radical production

during DENA and CCl4 metabolism (Prince., 2004).

On the other hand, the treatment of HCC has suggested in recent years via chemotherapies. Anticancer

drugs are widely used against variety of human cancer. However, while they also exhibit severe

toxicity and undesirable side effects (Minami et al., 2010). Adriamycin is one of anticancer drug. Two

main mechanisms are proposed to explain the adriamycin cytotoxicity. One suggests that the drug is

bio activated by NADPH cytochrome-P450-reductase and interacts with DNA leading to the inhibition

of both replication and transcription of DNA (Sawamura et al., 1996).Also it can serve as an electron

acceptor from microsomal and nuclear flavoproteins. The other mechanism suggests that adriamycin

induces an oxygen free radical formation causing an oxidative stress in cells and hence nucleic acid

cleavage (Feinstein et al., 1993; Stewart and Ratain, 2001).

There are a number of evidences indicating that natural substances from edible and medicinal plants

exhibited strong antioxidant activity that could act against hepatic toxicity caused by various toxicants

(Hassan et al., 2014 ; Othman et al., 2014). One of those candidate plants is cape gooseberry that has

various bioactive compounds (withanolides and phenolics) (Fang et al., 2012). Some of these

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compounds have a strong antioxidant property and prevent peroxidative damage to liver microsomes

and hepatocytes (Wang, et al., 1999; Hassan and Abdel-Aziz, 2010). Natural antioxidants could

prevent the deleterious effects of toxic agents by scavenging free radicals and other reactive oxygen

species or by modulation of the inflammatory response. Liver-protective herbal drugs contain a variety

of chemical constituents like phenols, coumarins, lignans, essential oil, monoterpenes, carotinoids,

glycosides, flavanoids, organic acids, lipids, alkaloids and xanthones derivatives (Qiu et al., 2007and

Sindhu et al., 2012). The present decreased of TG, LDL-c and VLDL-c in HCC rats received cape

gooseberry may be due to effects of cape gooseberry on lipid profile and HDL-c could be related to

antioxidant activity which might attribute to those identified compounds of cape gooseberry,

flavones, alkaloids, (Osho et al., 2010 ; Osman et al., 2013). Various evidence indicates that many

medicinal plants have been found to be useful to successfully manage hyperlipidemia (Lin et al., 2005;

Rashwan, 2012).This is probably due to the wealth of phytosterols in the cape gooseberry fruit which

induce a decrease in lipoprotein cholesterol levels in total plasma (Ramadan et al., 2011). The

observed decreased of cholesterol in HCC+ cape gooseberry rats may be due to due to the

hypocholesterolemic effects of cape gooseberry are mainly due to the lycopene existing in the plant

which is a strong antioxidant which inhibits the production of LDL and presumably increases the

escritoires through releasing cholesterol; therefore, it reduces blood cholesterol level and controls

cholesterol synthesis (Zarei et al., 2011; Ramadan, 2012). A significant an increase of total protein at

cape gooseberry supplementation may be due to cape gooseberry rich in phenolic compounds and

flavonoids that are widely used as antioxidant (Abd El-Ghany and Nanees, 2010), or may be due to

cape gooseberry contains zeaxanthin and beta cryptoxanthin esters or carotenoid esters which can be

used as food additives or nutraceuticals (Pintea et al., 2005). Also, the decreased of total bilirubin

(Tb) in HCC+ cape gooseberry rats may be due to the prevention of the leakage of intracellular

enzymes by its membrane stabilizing activity according to Chang et al. (2008 ;Osman et al., 2013).

An amelioration of AFP after supplementation of cape gooseberry juice to HCC rats may be attributed

to CG antioxidant activity. Additionally, the observed improvement of the occurred alterations in liver

enzymes including ALT, AST, ALP, and γ-GT in HCC+ cape gooseberry rat groups may be due to

alterations by cape gooseberry juice is a clear indication of the improvement of the functional status of

hepatocytes with preservation of cellular architecture leakage of intracellular enzymes by its

membrane stabilizing activity (Chang et al., 2008; Tatiya et al., 2012; Al-Olayan et al., 2014). The

successful of CG supplementation to HCC rats model in amelioration of oxidative stress and

improvement of antioxidants defense mechanism is consider a good indicator for its anti-lipid

peroxidative property and antioxidant activity through its high levels of antioxidants compounds such

Vol 22, No. 11;Nov 2015


as polyphenols and other like flavonoids ( Abdel Moneim and El-Deib, 2012 ; Hassan and Ghoneim,

2013). Also the observed decreased of MDA and NO in HCC rats received cape gooseberry may be

due to free radicals scavengers, potential mechanism by which cape gooseberry juice can act as

anti-inflammatory as well as the inhibition of the induction of inducible nitric oxide synthase (iNOS)

protein/enzyme and thus protect the liver. Therefore, dietary consumption of cape gooseberry extract

may be used as best potential antioxidant and hepatoprotective in liver toxicity (Rashwan, 2012;

El-Gengaihi et al., 2013). NO plays crucial roles in inflammation and liver injury (Leung et al., 2011).

It is produced in large quantities by Kupffer cells, endothelial cells, and the hepatocytes themselves in

response to tissue damage and inflammation induced by various xenobiotics including CCl4. In

addition, its role in oxidative stress cannot be neglected, since high levels of NO have been associated

with oxidative injury via lipid peroxide (Breikaa et al., 2013; Al-Olayan et al., 2014). Also the

decreased of AO and XO in cape gooseberry administered rat may be due to the presence of the

active compounds in CG that have biological significance in the elimination of reactive free radicals,

aldehydes are highly reactive molecules that may have a variety of effects on biological systems. They

can be generated from a virtually limitless number of endogenous and exogenous sources. Although

some aldehyde-mediated effects such as vision are beneficial, many effects are deleterious, including

cytotoxicity, mutagenicity, and carcinogenicity. A variety of enzymes have evolved to metabolize

aldehydes to less reactive forms. Changes in aldehydes activity have been observed during

experimental liver carcinogenesis and in a number of human tumors, including some liver, colon, and

mammary cancers. Changes in aldehydes define at least one population of preneoplastic cells having a

high probability of progressing to overt neoplasms (Wu et al., 2005; Saada et al., 2010; Osman et al.,

2013). Cape gooseberry supplementation showed a significant antioxidant status as manifested by

elevation of GSH, TAC, SOD, CAT and GSH-PX ase. Many plant secondary metabolites act as potent

antioxidants have shown that free radical scavenger/antioxidant such as superoxide dismutase (SOD),

catalase (CAT), reduced glutathione (GSH) and glutathione peroxidase (GPX), reduce and prevent

the tissue damage induced by different hepatotoxins (Hassan et al., 2014). The first line of defense

against superoxide anion (O2), H2O2 and (OH -)are the major ROS, which induce, cell degeneration by

increasing LPO of cell membrane lipids are SOD, CAT GPX are the family of metallo enzymes that

convert O2- to H2O2. The toxic end products of peroxidation induce damage of the structural and

functional integrity of cell membranes, break DNA strands and denature cellular proteins. The natural

cellular antioxidant enzymes include SOD, is an important enzyme as because it is found virtually in

all aerobic organisms. O2- is the only known substrate for SOD and it is considered to be a stress

protein, which is synthesized in response to oxidative stress. It scavenges superoxide radicals by

Vol 22, No. 11;Nov 2015


speeding up their dismutation (Kanimozhi and Prasad, 2009; Vásquez-Garzón et al., 2009).

In conclusion, the present data indicated the efficacy of CG juice supplementation as an

anti-hepatocellular carcinoma in addition to its ability as a chemosensitizer for ADR treatment. This is

mediated by intracellular pathways, involving improvement the alterations in liver functions as well as

other aspects of HCC, the suppression of oxidative stress and modulation of antioxidant defense

mechanism. Thus, supplementation with edible CG may help in safe application of cancer technology

in medicine as well as in many other aspects of nowadays life. Fractionation guided evaluation could

help in the development of ideal anticancer in the near future.

References

Abd El-Ghany M A and Nanees Y E M (2010) Effect of marjoram leaves on injured liver in

experimental rats. Report Opinion 2(12): 181-191.

Abdel Moneim A E and El-Deib K M (2012) The Possible protective effects of Physalis peruviana on

carbon tetrachloride-induced nephrotoxicity in male albino rats. Life Sci J 9(3): 1038-1052.

Acosta A A M D (1983) Directimmunoenzymatic determination of AFP in serum or plasma. J Clin

immunoassays 6: 41.

Aebi H (1984) colorimetric method for determination of catalase. Methods Enzymol 105:121-126.

Alberts D S and Salmon S E (1975) Adriamycin (NSC 123127) in the treatment of alkylator -resistant

multiple myeloma: a pilot study. Cancer Chemother Rep 59: 345-350.

Allain C C Poon L S Chan C S Richmond W Fu P C (1974) Enzymatic determination of total serum

cholesterol. Clin Chem 20: 470-475.

Al-Olayan E El-Khadragy M FOthman MS Aref A Kassab R and Abdel Moneim AE (2014) The

potential protective effect of Physalis peruviana L against carbon tetrachloride induced hepatotoxicity in

rats is mediated by suppression of oxidative stress and down regulation of MMP-9 expression. Oxidative

Medici Cellular Longevity Article ID 381413, 12 pageshttp://dx.doi.org/10.1155/381413

Anwar W A Khaled H M Amra H A El-Nezami H and Loffredo C A (2008) Changing pattern of

hepatocellular carcinoma (HCC) and its risk factors in Egypt possibilities for prevention. Mutat Res

659:176-184.

Vol 22, No. 11;Nov 2015


Arun M and Asha V V (2007) Preliminary studies on anti-hepatotoxic effect of Physalis peruviana

(Solanaceae) against carbon tetrachloride induced acute liver injury in rats. J Ethnopharmacol 111:

110-114.

ArzumanyanA Reis H M G PV and FeitelsonM A (2013) Pathogenic mechanisms in HBV and HCV

associated hepatocellular carcinoma. Nature Reviews Cancer vol. 13: pp 123–135.

Bagchi D Bagchi M and Stohs SJ (1997) Comparative in vitro oxygen radical scavenging ability of zinc

methionine and selected zinc salts and antioxidants. Gen Pharm 28:85–91.

Bailey T Berg Pand Sandy C (2001)The effect of high performance work practices on employee

earnings in the steel apparel and medical electronics and imaging industries. Industrial Labor Relations

Review54(2A): 525–543.

Belfield A and Goldberg DM (1971) Normal ranges and diagnostic value of serum 5,nucleotidase and

alkaline phosphatase activities in infancy. Arch Dis Child 46: 842– 846.

Benzie I F F (2003) Evaluation of dietary antioxidants. Comp Bio Chem Physiol 136: 113-126.

Beutler E Duron O and Kelly M B (1963) Colorimetric method for determination of GSH. J Lab Clin

Med 61: 882–888.

Breikaa R M Algandaby M M El-Demerdash E Dhanapakiam P Joseph J M Pamaswany V K and

Abdel-NaimA B (2013) Biochanin protects against acute CCl included hepatoxicity in rats. 4 Bioscoi

Biotechnol Biochem 77: 909-916.

Bruix J and Sherman M (2005) Management of hepatocellular carcinoma. An update Hepatology 42:

1208-1236.

Carr B I and Nagalla S (2010) In hepatocellular carcinoma. Diagnosis and treatment, Brain Carr (ED)

Humana Press New York USApp 527–568.

Chang J C Lin CC Wu SJ Lin DL Wang SS Miaw CL and Ng LT (2008)Antioxidative and

hepatoprotective effects of Physalis peruviana extract against acetaminophen induced liver injury in

rats. Pharm Bio 46(10-11): 724-731.

Ciriolo M R (2005) Redox control of apoptosis. Antioxid Redox Signal 7: 432-435.

Vol 22, No. 11;Nov 2015


Cooper RG and Magwere T (2008) Chloroquine novel uses and manifestations Indian. J Med Res 127:

305–316.

Cui WW Saint-Amant L and Kuwada JY (2004) Shocked gene is required for the function of a premotor

network in the zebrafish CNS. J Neurophysiol 92: 2898-2908.

Dakshayani K B Subramanian P and Manivasagam T (2005) Imbalance melatonin modulates the

oxidantantioxidant during N-nitrosodiethy-lamine induced hepatocarcinogenesis in rats. J Pharm Sci 8:

316-321.

De Duve C and Baudhuin P (1996) Peroxisomes (microbodies and related particles).Physiol Rev 46:

323–357.

Dewa Y Nishimura J Muguruma M Jin M Kawai M Saegusa Y Okamura T Umemura T and Mitsumori

K (2009) Involvement of oxidative stress in hepatocellular tumor-promoting activity of oxfendazole in

rats. Arch Toxicol 83: 503–511.

Di Bisceglie A M Sterling R K Chung RT Everhart J E Dienstag J L Bonkovsky H L Wright E C Ever-

son GT Lindsay K LLok A S Lee W M Morgan T R Ghany M G and Gretch D R(2005) Serum

alpha-fetoprotein levels in patients with advanced hepatitis C Results from the HALT-C Trial. J

Hepatology 43: 434-441.

DiMarco A Gaetani M and Scarpinato B (1969) Adriamycin (NSC-123,127) a new antibiotic with

antitumouractivity. Cancer Chemother Rep 53(1): 33-37.

Dinan L Sarker S and Sik V (1997) 28-Hydroxywithanolide e from Physalis peruviana. Photochemistry

44: 509−512.

DiSilvestro RA (2000) Zinc in relation to diabetes and oxidative disease. J Nutr130:S1509 –S1511.

El shahat A N (2013) Efficiency of broccoli in attenuating of some biochemical disorders in rats

exposed to γ-irradiation. Arab J Nuc Sci Applic 46 (4): 260-267.

El-Gengaihi S E Hamed M AKhalaf-Allah R and Mohammed M A (2013) Golden berry juice attenuates

the severity of hepatorenal injury. J Dietary Supplements 10:(4): 357–369.

El-Serag HB (2011) Hepatocellular carcinoma. N Engl J Med 365(12): 1118-1127.

Vol 22, No. 11;Nov 2015


Fang ST Liu JK and Li B (2012) Ten new withanolides from Physalis peruviana Steroids. 77: (1-2):

36–44.

Farombi E O Shrotriya S and Surh Y J (2009) Kolaviron inhibits dimethyl nitrosamine-induced liver

injury by suppressing COX-2 and iNOS expression via NF-κB and AP-1. Life Sci 84: 149–155.

Fassati P and Prencipe L (1982) Serum triglycerides determined colorimetrically with an enzyme that

produces hydrogen peroxide. Clin Chem 28(10):2077-2080.

Feinstein E Canaani E and Weiner L M (1993) Dependence of nucleic acid degradation on in situ

free-radical production by Adriamycin. Biochemistry 44: 210–215.

FialaS andFiala E S (1973)Activation by chemical carcinogens of gamma-glutamyl transpeptidase in rat

and mouse liver. J Natr Cancer Inst Jul 51 (1):151–158.

Friedewald W T Levy R I and Fredrickson D S (1972) Estimation of the concentration of low density

lipoprotein cholesterol in plasma, without use of the preparative ultracentrifuge. Clin Chem 18: 499-502.

Gordon A H Green D E and Subrahmanyan V (1940) Liver aldehydeoxidase. Biochem J 34 (5):

764–774.

Griffith O W Meister A (1980) Excretion of cysteine and gamma-glutamylcysteine moieties in human

and experimental animal gamma-glutamyl transpeptidase deficiency. ProcNatlAcadSci U S A Jun

77(6): 3384–3387.

Hannigan M H and Pitot H C (1985) Gamma-glutamyltranspeptidase and mdashIts role in

epatocarcinogenesis. Carcinogenesis 6: 165-172.

Hassan HA Edress G M El-Gamal E El-SayedE A (2014) Amelioration of cisplatin-induced

nephrotoxicity by grape seed proanthocynidin extract and fish oil is mediated by lowering oxidative

stress and DNA damage. Cytotechnology 66: 419–429.

Hassan A I and Ghoneim M A M (2013) A possible Inhibitory effect of (Physalispubescens) on diabetes

in male rats. World Appl Sci J 21 (5): 681-688.

Hassan H A Abdel-Aziz A F (2010) Evaluation of free radical- scavenging and anti-oxidant properties of

black berry against fluoride toxicity in rats. Food Chem. Toxicol 48: 1999-2004.

Vol 22, No. 11;Nov 2015


Hayakawa R and Jap J (1961) Estimation of zinc. Toxicol Environ Health 8: 14-18.

Helvaci S Kökdil G Kawai M Duran N Duran G and Güvenc A (2010) Antimicrobial activity of the

extracts and physalin D from Physalisalkekengi and evaluation of antioxidant potential of Physalin D.

Pharm Biol 48(2):142-150.

Henry R J (1964) Determination of total protein by colorimetric method.Clinical Chemistry Harper

and Row publishers New York.P181.

Hille R (2005) Molybdenum-containing hydroxylases. Arch BiochemBiophys 433 (1): 107–116.

Jemal A Murray T Ward E Samuels A TiwariR CGhafoor A Feuer E J Thun M J Cancer J and Clin

C A (2005) Cancer statistics. 55(1):10-30.

JemalA Bray F Center M MFerlay J Ward E and Forman D (2011) Global cancer statisticsCA Cancer. J

Clinicians61:(2): 69–90.

Jendrassik L and Grof N (1938) In-vitro determination of total and direct bilirubin in serum. J Biochem

299: 81- 88.

Jiang H E Li X Zhao Y X Ferguson D K HueberF Bera S Wang Y F Zhao L C Liu C J and Li C S

(2006) A new insight into Cannabis sativa (Cannabaceae) utilization from 2500-year-old Yanghai

Tombs Xinjiang China. J Ethnopharmacology 108: 414-422.

Johnson C StubleyBeedham C and Stell J G P (1984) Elevation of molybdenum hydroxylase levels in

rabbit liver after ingestion of phthalazine or its hydroxylatedmetabolite. BiochemPharmacol 33:

3699-3705.

Kaefer C M and Milner J A (2008)The role of herbs and spices in cancer prevention. J NutrBiochem

19:347–361.

Kanimozhi P and Prasad R N (2009) Antioxidant potential of sesamol and its role on radiationinduced

DNA damage in whole-body irradiated Swiss albino mice. Environ ToxicolPharmacol 28(2): 192–197.

Kawasaki N Hamamoto Y Nakajima T Irie K Ozawa H and Periodontal S (2004) Regeneration of

transplanted rat molars after cryopreservation. Arch Oral Biol 49: 59-69.

Vol 22, No. 11;Nov 2015


Kobayashi T and Kawakubo T (1994) Prospective investigation of tumor markers and risk assessment in

early cancer screening. Cancer 73: 1946-19S3.

Koracevic D Koracevic G Djordjevic V Andrejevic S and Cosic V (2001) Method for the measurement

of antioxidant activity in human fluids.J ClinPathol 54:356 – 361.

Kregel K C and Zhang H J (2007)An integrated view of oxidative stress in aging basic mechanisms

functional effects and pathological considerations. Am J PhysiolRegulIntegr Comp Physiol 292: 18–36.

Lai C L Lok A S F Wu P C Chan G C B and Lin H J (1988)Doxorubicin versus no antitumor therapy in

inoperable hepatocellular carcinoma a prospective randomized trial. Cancer 62: 479-483.

Leung T M Fung M L LiongE CLau T Y H Nanji A A and Tipoe G L (2011) Role of nitric oxide in

the regulation of fibrogenic factors in experimental liver fibrosis in mice. Histology and Histopathology

26(2): 201–211.

Linder S (1995)Foliar analysis for detecting and correcting nutrient imbalances in norway spruce.

Ecological Bulletins (Copenhagen) 44: 178-190.

Lin C C Li T C and Lai M M (2005) Efficacy and safety of monascuspurpureus Went rice in subjects

with hyperlipidemia.Eur J Endocrinol 153(5): 679-686.

Lopez–Virella M F (1977) Enzymatic colorimetric method for determination of HDL .Clin chem 23:

882.

Luczaj W and Skrzydlewska E (2003) DNA damage caused by lipid peroxidation product. Cellular and

molecular Bio 8(2): 391-413.

MacPhee D G (1998) Time-dependent mutagenesis and cancer a new role for antimutagenesis in cancer

prevention. Mutat Res 402:29–39.

Maideen N M Velayutham R and Manavalan G (2012) Activity of prosopis cineraria against

N-nitrosodiethylamine induced liver tumors by regulating the levels of tumor marker lipid peroxidation

and antioxidants. AJPLS 2: 1-9.

Manimaran A and Rajneesh C P (2009) Activites of antioxidant enzyme and lipid peroxidation in

ovarian cancer patients. Academic J Res 2(2): 68-72.

Vol 22, No. 11;Nov 2015


Minami M Matsumoto S and Horiu chi H (2010) Cardiovascular side-effects of modern cancer therapy.

Circ J 74(9): 1779-1786.

Montgomery H A C and Dymock J F (1961) The determination of nitrate in water. Analyst 86: 414-416.

Niskikimi M Rao N A and Yog K (1972)Colormetric determination of superoxide dismutase activity.

BiochemBiophys Res Commun 46: 849-851.

Ogawa T (1985) Studies on endoscopic local injection of an anticancer agent into experimental canine

stomach cancer. Kitakant Med J 35: 463-476.

Ohkawa H Wakatsuki A and Kaneda C (1982) Assay for lipid peroxides in animal tissues by

thiobarbaturic acid reaction. Anal Biochem 95: 351-358.

Olubunmi BO Adewumi TA Oladayo AJ (2011) Antihepatotoxic effects of aqueous extract of the

leaves of MorindalucidaBenth on ethanol-induced hepatotoxicity in rabbit. Austr J Basic ApplSci 5:

285-291.

Oshoa Adetunji T Fayemi S O and Moronkola D P (2010) Antimicrobial activity of essential oils

of Physalis angulate L.J Home 7: 303-306.

Osman N N AL-seeni M N Alkhatib M H and Al-shreef H A (2013) Modulation of radiation injury by

Physalis peruviana.Life Sci J 10(4):3403-3410.

Othman M S Safwat G Aboulkhair M and AbdelMoneim A E (2014) The potential effect of berberine in

mercury-induced hepatorenal toxicity in albino rats. Food Chemical Toxicology 69: 175-181.

Paglia D E and Valentine W N (1967) UV method for determination of glutathione peroxidase. J Lab

Clin Med 70: 158-169.

Pasupathi M McLean K C and Weeks T (2009) To tell or not to tell disclosure and the narrative self. J

Pers 77: 1–3510.

Patel N H Condron B G and Zinn K (1994) Pair-rule expression patterns of even-skipped are found in

both short- and long-germ beetles nature. 367: 429-434.

Vol 22, No. 11;Nov 2015


Pintea A Varga A Stepnowski P Socaciu C Culea Mand Diehl HA (2005) Chromatographic analysis

ofcarotenol fatty acid esters in Physalisalkekengi andHippophaerhamnoides.Phytochem Anal 16:

188-195.

Prince J T (2004) The need for a public proteomics repository. NatrBiotechnol22: 471–472 .

Pulpanova J Kovarova H and Ledvina M (1982) Changes of the reduced glutathione concentration and

the activity of glutathione reductase in some tissue of rats gamma irradiated after the administration of

cystamine. Radiobiol 30 (3): 413.

Qiu L Jiang Z H Liu H X Chen L X Qu G X and Qiu F (2007) Flavonoid glycosides of the Calyx

Physalis. J Shenyang Pharm Uni 24(12): 744-747.

Ramadan M F (2012) Physalis peruviana pomace suppresses high-cholesterol diet-induced

hypercholesterolemia in rats. Grasasy Aceites 63 (4): 411-422.

RamadanM F Zayed R Abozid M and Asker M M S (2011) Apricot and pumpkin oils reduce plasma

cholesterol and triacylglycerol concentrations in rats fed a high fat diet. Grasas Aceites 62: 443-452.

Ramakrishnan G Raghavendran H R Vinodhkumar R and Devaki T (2006) Suppression of

N-nitrosodiethylamine induced hepatocarcinogenesis by silymarin in rats. Chem Biol Interact 161:

104–114.

Rashwan N M (2012) Effect of Physalis and choline on lipid profile and antioxidant activity in hepatic.

Austr J Basic ApplSci 6(8): 654-660.

Reddy C V K Sreeramulu D and Raghunath M (2010) Antioxidant activity of fresh and dry fruits

commonly consumed in India. Food Res Inte 43: 285−288.

Regnstrom J Nilsson J Tornvall PLandou C and Hamsten A (1992) Susceptibility to low-density

lipoprotein oxidation and coronary atherosclerosis in man. Lancet 339: 1183—1186 .

Reitman S and Frankel S (1957) A colorimetric method for the determination of serum glutamic

oxaloacetic and glutamic pyruvic transaminase. J Clin Pathol 28: 56-63.

Robak J and Glyglewsi R J (1988) Flavonoids are scavengers of superoxide anions. Biochem. Pharmacol

37: 837–841.

Vol 22, No. 11;Nov 2015


Rocchi C A Bizzarri R and Cannistraro Ì (1997) Water residence times around copper plastocyanin: A

molecular dynamics simulation approach. Chemical Physics 214: 261-276.

Roughead ZK Johnson LK and Hunt JR (1999) Dietary copper primarily affects antioxidant capacity and

dietary iron mainly affects iron status in a surface response study of female rats fed varying

concentrations of iron zincand copper. J Nutr129:1368 –1376.

Saada H N Rezk R G and Eltahawy N A (2010) Lycopene protects the structure of the small intestine

against gamma-radiation-induced oxidative stress. Phytother Res 24: S204–S208.

Sako Y Hosoi-Tanabe S and Uchida D (2004) Fluorescence in situ hybridization using rRNA-targeted

probes for simple and rapid identification of the toxic dinofla-gellates Alexandriumtamarense and A.

catenella J Phycol 40: 598–605.

Sakr S A Mahran H A and Lamfon H A (2011) Protective effect of ginger (Zingiberofficinale) on

adriamycin - induced hepatotoxicity in albino rats. J Medic Plants Res 5(1): 133-140.

Sarkar V A Basak R Bishayee ABasak J and Chatterjee M (1997) B-carotene inhibits rat liver

chromosomal aberrations and DNA chain break after a single injection of diethylnitrosamine. Br J

Cancer 76: 855-861.

Satheesh M and Pari L (2008) Effect of pterostilbene on lipids and lipid profiles in

streptozotocin–nicotinamide induced type 2 diabetes mellitus. J Appl Biomed 6: 31–37.

Sawamura A O Aoyama TTamakoshiK Mizuno K Suganuma N Kikkawa F and Tomoda Y

(1996) Transfection of humancytochrome P-450 reductasec DNA and its effect on the sensitivity to

toxins. Oncology 53: 406–411.

Schumann G Bonora R Ceriotti F Férard G Ferrero C A Franck P F H Gella F J Hoelzel W Jørgensen P

J Kanno T Kessner A Klauke R Kristiansen N Lessinger J M Linsinger T P J Misaki H Panteghini M

Pauwels J Schiele F and Schimmel H G (2002) 725 IFCC primaryreference procedures for the

measurement of catalytic activity concentrations of enzymes at 37oC part 5 Clin. Chem Lab Med 40:

725-733.

Schwartz M K and Bodansky 0 (1965) Serum 5-nucleotidase in patients with cancer. Cancer 18:

886-892.

Vol 22, No. 11;Nov 2015


Sell S and Becker F F (1978) Alpha-Fetoprotein. J Natl Cancer Inst 60: 19–26.

Sen A (1995)`Rationality and social choice'. AmEconom Review 85: 1-24.

Seufi A M Ibrahim S SElmaghraby T K and Hafez E E (2009)Preventive effect of the

flavonoidquercetin, on hepatic cancer in rats via oxidant antioxidant activity molecular and histological

evidences. J Exp Clin Cancer Res 28: 80.

Shaarawy S MTohamyA AElgendy S M Elmageed Z Y A Bahnasy A Mohamed M S Kandil E

andMatrougui K (2009) Protective effects of garlic and silymarin on NDEA-induced rat hepatotoxicity.

Int J Bio Sci 5 (6): 549-557.

Shariff N Sudarshana M S Umesha S and Hariprasad P (2006) Antimicrobial activity of

rauvolfiatetraphylla and Physalisminima leaf and callus extracts. African J Biotech 5: 946-950.

Sindhu G Susithra M and Vijayalakshmi N (2012) Evaluation of hepatoprotective effect of berberin in

paracetamol induced experimiental hepatotoxicity in wistar rats. Int J Recent Sci Res 3(7): 569-573.

Singha B N Braj R Singh B R Sarma B K and Singha H B (2009) Potential chemoprevention of

N-nitrosodiethylamine-induced hepatocarcinogenesis by polyphenolics from acacia niloticabark.Chem

Bio Interact 181: 20–28.

Stewart C F and Ratain M J (2001)Chapter 19: Pharmacology of cancer chemotherapy, 19 6

topoisomerase interactive agents. In DeVitta V Jr Hellman S Rosenberg S A (Eds.) Cancer Principles

and Practice of Oncology on CD-ROM 6th edLippincott Williams and Wilkins Philadelphia pp 1–16.

Stripe F and Della Corte E (1969) The regulation of rat liver xanthine oxidase. J BiolChem 244:

3855-3863.

Subramanian P Mirunalini S Dakshayani K B Pandi-Perumal S R Trakht I and Cardinali D P (2007)

Prevention by melatonin of hepatocarcinogenesis in rats injected with N-nitrosodiethylamine. J Pineal

Res 43: 305–312.

Szasz G A (1969) kinetic photometric method for serum 13 γ-Glutamyltranspeptidase.ClinChem 15:

124 - 136.

Szefer P and Nriagu J (2007) Mineral components in foods .New York CRC P 480.

Vol 22, No. 11;Nov 2015


Tang Q Y Yao D F and Lu J X (1999)Expression and alterations of different molecular form

gammaglutamyltransferase and total RNA concentration during the carcinogenesis of rat hepatoma.

World J Gastroenterol 5: 356–358.

Tatiya A U Surana S J Sutar M P and Gamit N H (2012)Hepatoprotective effect of poly herbal

formulation against various hepatotoxic agents in rats Pharmacognosy. Res 4)1:(50–56.

Thangaraju M Rameshbabu H Vasavi S Ilanchezian RVinitha and Sachanandam P (1998) The

salubrious effect of tamoxifen on serum marker enzymes glycoproteinslysosomal enzyme level in breast

cancer women. Mol Cell Biochem 185: 85-94.

Thomson C D (1998) Selenium speciation in human body fluids. Analyst 123: 827-831.

Thusu N Raina P N and Johri R K (1991) γ-Glutamyltranspeptidse activity in mice Age dependent

changes and effect of cortisol. Ind J ExpBiol 29: 1124-1126.

Vanisree A J and C S Shyamaladevi (1998) Effect of therapeutic strategy established by N-acetyl

cysteine and vitamin C on the activities of tumor marker enzymes in vitro. Indian J Pharmacol 31:

275-278.

Vásquez-Garzón V R Arellanes-Robledo R García-Román D I Aparicio-Bautista S and Villa-Treviño J

(2009) Inhibition of reactive oxygen species and pre-neoplastic lesions by quercetin through an

antioxidant defense mechanism. Free Radic Res 28: 37-43.

Ventura S and king E J (1951) Determination of serum copper by sodium diethyldithiocarbamate

method. Biochem J 48: 66.

Waller R A and Duncan D B (1969) Bayes rule for the symmetric multiple comparision problems. Am J

Stat Assoc 64: 1484-1503.

Wang I K Lin-Shiau S Y and Lin J K (1999) Induction of apoptosis by apigenin and related flavonoids

through cytochrome c release and activation of caspase-9 and caspase-3 in leukaremia HL-60 cells.

Eurpean J Cancer 35: 1517−1525.

Wang J J Cortes E Sink L F and Holland J F (1971) Therapeutic effect and toxicity of adriamycin in

patients with nepotistic diseases. Cancer 28(4): 837-843.

Vol 22, No. 11;Nov 2015


Wang X Q Ongkeko W M Chen L Yang Z F Lu P Chen K K Lopez J P Poon R T and Fan S T

(2010)Octamer 4 (Oct4) mediates chemotherapeutic drug resistance in liver cancer cells through a

potential Oct4- AKT-ATP-binding cassette G2 pathway. Hepatology 52: 528-539.

Wattenberg L W (1990) Chemoprevention of cancer by naturally occurring and synthetic compounds.

Proc Assoc Cancer Res 32: 461-463.

Wolf P L (1999) Biochemical diagnosis of liver diseases. Ind J Clin Biochem 14: 59-90.

Wu S J Ng L T Huang Y M Lin D L Wang S S Huang S N and Lin C C (2005) Antioxidant activities of

Physalis peruviana. Biol Pharm Bull 28: 963–966.

Yao D F Dong Z Z and Yao D B (2004) Abnormal expression of hepatoma-derived gammaglutamyl

transferase subtyping and its early alteration for carcinogenesis of hepatocytes. Hepatobiliary Pancreat

Dis Int 3: 564–570.

Zarei O Irajian G R and Zarnani A H (2011) Peptide-based polyclonal antibody production against P110

protein of Mycoplasma genitalium. Avicenna J Med Biotech 3: 79-86.

Zhang Y JDeng G F Xu X R Wu S Li S and Li H B (2013) Chemical components and bioactivities of

cape gooseberry (Physalis peruviana). Int J Food Nutr Safety 3(1): 15-24.

Zollner N and Kirsch K (1962) Uber die quantitative bestimmung von lipoiden (Mikromethode)

mittels de vielennatiirlichen Lipoiden (allenbekannten Plasmalipoiden) gemeinsamen sulphophospho-

vanillinreaktion Z. Gesexp Med 135: 545-561.

Vol 22, No. 11;Nov 2015


Table 1: Serum and hepatic biochemical parameters in control and different treated rat groups.

Results are presented as means ±E (n=6 for each group).

C: Control, HCC: hepatocellular carcinoma, HCC+ADR: hepatocellular carcinoma+ Adriamycin,

HCC+CG: hepatocellular carcinoma+ Cape gooseberry, HCC+ADR+CG: hepatocellular carcinoma+

Adriamycin + Cape gooseberry. Mean with different letters (a- e) are significantly difference. Mean with the same letters are non-significantly difference.

Animal groups

Serum

HCC+A+CG HCC+CG HCC+A HCC C Parameters

512.09

±0.52e

498.1

±0.55d

610.29

±0.72c

768.6

±0.65b

490.69

±0.35a

Mean

± SE

TL

mg/dl

118.52

±1.12e

110.96

±1.14d

126.61

±1.12c

136.0

±1.16b

102.45

±1.05a

Mean

± SE

TC

mg/dl

88.27

±0.69d

86.79

±0.78ad

96.1

±0.76c

126.6

±0.8b

84.31

±0.71a

Mean

± SE

TG

mg/dl

32.52

±0.81ac

33.91

±0.81ac

31.84

±0.79c

25.08

±0.43b

34.67

±0.98a

Mean

± SE

HDL

mg/dl

68.35

±1.16e

59.21

±1.12d

75.57

±1.39c

85.66

±1.44b

50.86

±1.08a

Mean

± SE

LDL

mg/dl

17.65

±0.41a

17.35

±0.43a

19.22

±0.58a

25.32

±0.66b

16.92

±0.49a

Mean

± SE

VLDLmg/dl

6.28

± 0.55ed

6.35

± 0.48d

4.83

± 0.3cb

4.45

±0.2b

7.3

± 0.7a

Mean

± SE

TP

(mg/dl)

1

± 0.03ab

0.95

±0.02a

1.1

± 0.03ab

1.23

±0.06b

0.93

±0.01a

Mean

± SE

T-bilirubin

mg/dl

95.55

± 0.88d

100.65

± 0.86d

80.09

± 0.62c

71.85

± 0.52b

119.78

± 0.93a

Mean

± SE

Zn

(µg/ml)

295.8

± 0.63ac

303.43

± 0.55a

270.81

± 0.51cb

250.84

± 0.25b

310.79

± 0.53a

Mean

± SE

Cu

(µg/ml)

30.27

±0.52d

28.46

±0.15d

35.82

±0.82c

55.08

±1.15b

24.03

±0.46a Mean± SE

TL mg/g wet

tissue

Liver 120.88

±0.59d

118.57

±0.36a

123.13

±0.36c

128.2

±0.49b

117.24

±0.59a

Mean

± SE

TC mg/g wet

tissue

52.19

±0.52ed

53.93

±0.53d

49.1

±0.38c

63.9

±0.68b

39.29

±0.38a

Mean

± SE

TG mg/g wet

tissue

1.29

±0.02dc

1.95

±0.05a

1.28

±0.03c

1.06

±0.01b

2.05

±0.06a

Mean

± SE

TP

g/g wet tissue

Vol 22, No. 11;Nov 2015


Table 2: AFP and liver function enzymes in control and different treated rat groups.

Animal groups Parameters

Serum

HCC+A+CG HCC+CG HCC+A HCC C

1.25

±0.66a

1

±0.94a

1.33

±0.7a

2.57

±0.28b

0.99

±0.1a

Mean

± SE

AFP

ng /ml

45

± 0.63d

41.83

±0.6ad

49.16

±0.7c

60.5

±0.99b

40.16

±0.62a

Mean

± SE

AST

units/ml

37.33

±1.05acd

33.16

±1.06ad

38

±1.51ac

50.5

±1.76b

34.66

±1.02a

Mean

± SE

ALT

U/ml

105.65

±0.38ec

97.56

±0.5ad

109.4

±0.56c

153.11

±1.68b

93.63

±0.38a

Mean

± SE

ALP

IU/L

28.97

±1.02dc

23.7

±0.41a

27.04

±0.67c

32.02

±1.07b

21.53

±0.5a

Mean

± SE

γ-GT

U/L

16.68

±0.31e

17.8

±0.28d

14.54

±0.21c

15.6

±0.23b

18.77

±0.34a

Mean

± SE

AST U/g wet

tissue

Liver

46.07

±0.54e

57.13

± 0.61d

42.74

±0.56cb

42.41

±0.51b

60.77

±0.67a

Mean

± SE

ALT U/g

wet tissue

40.16

±1.09ec

43.5

±1.1dc

42.5

±1.04c

54.9

±1.14b

33.03

±0.76a

Mean

± SE

ALP IU/g wet

tissue

27.72

±0.85dc

25.84

±0.79acd

26.72

±0.87cb

28.63

±0.89bd

23.82

±0.68a

Mean

± SE

γ-GT

U/g


C: Control, HCC: hepatocellular carcinoma, HCC+ADR: hepatocellular carcinoma+ Adriamycin, HCC+CG: hepatocellular

carcinoma+ Cape gooseberry, HCC+ADR+CG: hepatocellular carcinoma+ Adriamycin+ Cape gooseberry. Mean with different

letters (a- e) are significantly difference. Mean with the same letters are non-significantly difference.

Vol 22, No. 11;Nov 2015


Table 3: Hepatic oxidative stress and antioxidant biomarkers in control and different treated rat

groups.


C: Control, HCC: hepatocellular carcinoma, HCC+ADR: hepatocellular carcinoma+ Adriamycin, HCC+CG: hepatocellular

carcinoma+ Cape gooseberry, HCC+ADR+CG: hepatocellular carcinoma+ Adriamycin+ Cape gooseberry. Mean with different

letters (a- e) are significantly difference. Mean with the same letters are non-significantly difference.

Animal groups

Parameters

Liver

HCC+A+CG HCC+CG HCC+A HCC C

530.5 520.38 581.07 701.12 512.11 Mean

± SE

MDA

nmol/g wet tissue ±0.49a ±0.59a ±0.77c ±0.82b ±0.64a

626.04 611.3 652.81 724.41 590.16 Mean

± SE

NO

µmol/g ±0.81d ±0.78d ±0.89c ±0.88b ±0.72a

2.28

±0.042e

1.61

±0.03d

3.28

±0.04c

3.67

±0.06b

0.84

±0.02a

Mean

± SE

AO

umol/min

15.83

±0.47e

13.33

±0.45d

18.43

±0.66c

25.13

±0.95b

11

±0.28a

Mean

± SE

XO mmole/hour

/gm tissue

15.92 17.84 12.78 10.01 19.11 Mean

± SE

GSH mmol/g

wet issue ±0.41d ±0.44ad ±0.35cb ±0.26b ±0.53a

110.18 118.12 101.1 95.26 128.1 Mean

± SE

TAC

mM/g ±0.71d ±0.63d ±0.69cb ±0.47b ±0.78a

817.28 841.71 836.97 416.54 871.54 Mean

± SE

SOD

U/g wet tissue ±0.63a ± 0.79a ±0.73a ±0.42b ±0.88a

183.52 180.68 150.28 132.3 190.7 Mean

± SE

Catalase

U/g wet tissue ±0.91ed ±0.64d ±0.59c ±0.23b ±0.85a

781.44 797.08 760.95 745.04 801.94 Mean

± SE

GSH-PX

U/gt ±1.63e ±1.59d ±1.65c ±1.75b ±1.83a

Efficiency of cape gooseberry in attenuating some biochemical disorders and oxidative stress...

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