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THE PROTECTIVE EFFICACY OF VITAMINS (C AND E), SELENIUM AND

SILYMARIN SUPPLEMENTS AGAINST ALCOHOL TOXICITY

Shalan M.G.*, Abd Ali W. Dh., Shalan A.G.

 

*Al-Arish Fac. of Education, Suez Canal Univ., Biological and Geological Sciences Dpt., North Sinai, Egypt.

 

High Health Institute, Egdabia, Libya.

 

Port Said Fac. of Education, Suez Canal Univ., Biological and Geological Sciences Dpt., Port Said, Egypt

ABSTRACT: This study aimed at investigating the efficacy of vitamins (C and E), selenium and silymarin (an

 

antioxidant complex from Silybum marianum) supplementation in reducing toxic effects of ethanol on liver

 

 

 

weight and some blood parameters. Sixty male rabbits, individually housed in steel cages, were randomly

 

divided into three groups. The first was a control group, the second received balanced diet and daily 20% (v/

 

v) ethyl alcohol in their drinking water, the third received the same diet and 20% (v/v) ethanol in their drinking

 

water and treated with vitamin C (1 mg/100 g body weight, BW), vitamin E (1 mg/100 g BW), selenium (0.01 mg/

 

100 g BW) and silymarin (1 mg/100 g body weight) by gastric tube daily. Five animals per group were slaughtered

 

every two weeks and liver and blood samples were taken after 2, 4, 6 and 8 weeks of treatment. Ethanol

 

decreased body weight of rabbits and induced hepatomegally and apoptotic DNA fragmentation in hepatocytes.

 

Chronic alcohol consumption induced significant increases in serum glucose, triglycerides and cholesterol

 

levels whereas serum total protein content decreased. Significant increases in serum ALT, AST, ALP and LDH

 

activities were observed in ethanol-treated rabbits. The treatment of alcohol-abused animals with vitamins (C

 

and E), selenium and silymarin enhanced significant improvement in the biochemical, physiological and molecular

 

aspects indicating their protective effects against alcohol toxicity.

 

 

Key words: Rabbits; alcohol toxicity; vitamin C; vitamin E; selenium, silymarin.

 

 

W O R L D

 

RA B B I T

 

SCIENCE

 

 

 

INTRODUCTION

 

 

 

Alcohol consumption represents a large problem all over the world (Kumar and Clark, 2002). Alcohol

 

cannot be stored and obligatory oxidation must take place predominantly in the liver via alcohol

 

dehydrogenase. The production of potentially toxic acetaldehyde is enhanced and conversion to

 

acetate reduced (Sherlock and Dooley, 2002). The hydrogen produced replaces fatty acid as a fuel so

 

that fatty acids accumulate with consequent ketosis, triglyceridaemia, fatty liver and hyperlipidaemia

 

(Lieber, 1990). The conversion of alcohol to acetaldehyde also leads to inhibition of protein synthesis

 

 

(Bernal et al., 1992) and alter its metabolism (Apte et al., 2004). Ethanol was also reported to cause

 

 

 

many alterations in the activities of several enzymes under different nutritional conditions which

 

appeared to play a role in modulating this effect (Shalan, 1996). Ethanol was recorded to alter also

 

 

carbohydrate (Martin et al., 2004) and lipid (Ruf, 2004) metabolism.

 

Koyuturk et al. (2004) showed the protective effect of combination therapy with vitamins C and E,

 

and selenium on ethanol-induced duodenal mucosal injury. Marino et al. (2004) showed that vitamin

 

 

 

E protects against alcohol induced oxidative stress. The presence of selenium in combination with

 

 

vitamin E enhanced its activity in removing free radicals and prevented their formation (Saito et al.,

 

 

 

2003).

 

 

THE PROTECTIVE EFFICACY OF VITAMINS (C AND E), SELENIUM AND

 

SILYMARIN SUPPLEMENTS AGAINST ALCOHOL TOXICITY

 

 

Shalan M.G.*, Abd Ali W. Dh., Shalan A.G.

 

*Al-Arish Fac. of Education, Suez Canal Univ., Biological and Geological Sciences Dpt., North Sinai, Egypt.

 

High Health Institute, Egdabia, Libya.

 

Port Said Fac. of Education, Suez Canal Univ., Biological and Geological Sciences Dpt., Port Said, Egypt.

 

 

 

104

 

 

SHALAN et al.

 

Silymarin is an antioxidant flavonoid complex derived from the herb milk thistle (Silybum marianum).

 

 

 

It was proved to have a protective effect against experimental hepatotoxicity by regulating the

 

actions of the ultrastructures of the liver cells and improving the activities of hepato-cellular enzymes

 

 

and bile production (Hagymasi et al., 2002).

 

 

 

Thus the main purpose of our study was to investigate the protective effect of combined

 

supplementation with vitamins (C and E), selenium, and silymarin against alcohol toxicity.

 

 

MATERIAL AND METHODS

 

 

Animals and experimental treatments

 

 

 

Sixty male laboratory New Zealand White rabbits 8 weeks old (Oryctolagus cuniculus) weighing

 

 

 

(1000 ± 100 g) were housed individually in steel cages where laboratory balanced diet and water were

 

 

initially provided under standard controlled conditions (25 ± 2oC and relative humidity of 25 ± 5%).

 

 

 

Animals were randomly divided into three groups. The first group was normal controls (20 animals).

 

The second was ethanol group (20 animals). Each animal received 30 ml 20% (v/v) ethanol/day as

 

drinking water (absolute ethanol purchased from Al-Gomhoria Chemical Co., Egypt) and fed with

 

balanced diet. The third one was alcohol + antioxidant group (20 animals). Each animal received 30 ml

 

20% (v/v) ethanol/day as drinking water, fed with balanced diet and supplemented with 1 mg vitamin

 

C/100g body weight (BW), 1 mg vitamin E/100 g BW, 1 mg silymarin/100 g BW and 0.01 mg selenium/

 

100g BW by gastric tube daily.

 

 

Vitamin E (DL-a-tocopherol) and selenium (sodium selenite) were obtained from Merk (Darmstadt,

 

 

 

Germany). Silymarin was commercially available from Sedico Pharmaceutical Co. (Cairo, Egypt).

 

 

Sample collection and biochemical analyses

 

 

 

Animals were weighed at the beginning of the experiment and before each sample collection. Every

 

2 weeks 5 animals per group were anaesthetized and rapidly dissected. Livers were weighed

 

immediately after dissection.

 

Samples were collected after 2, 4, 6 and 8 weeks of alcohol consumption. Blood samples were collected

 

from the inferior vena cava in glass centrifuge tubes, then centrifuged for 15 min at 1000 g in a cooled

 

 

centrifuge (4oC). Sera were separated and stored at –30oC in deep freezer till further biochemical

 

 

 

measurements. Serum total protein, glucose, triglycerides, cholesterol, ALT (alanine aminotransferase),

 

AST (aspartate aminotransferase), ALP (alkaline phosphatase) and LDH (lactate dehydrogenase)

 

concentrations were determined automatically using Integra 800 auto-analyzer (Liver Institute,

 

Menoufiya University, Egypt).

 

 

Preparation of tissues, gel preparation and electrophoresis of lysate tissue

 

 

 

After dissection, liver was removed, blotted on filter paper and weighed. Portions of 10 mg were

 

 

taken immediately for gel examinations and the remaining portions were stored at –30oC.

 

 

 

Gels were prepared with 1.8 % electophoretic grade agarose (BRL). The agarose was boiled in Trisborate

 

 

EDTA buffer (1 × TBE buffer; 89 mM tris, 89 mM boric acid, 2 mM EDTA, pH 8.3). The 0.5 μg/

 

ml ethidium bromide was added to gel at 40oC. Gels were poured and allowed to solidify at room

 

 

 

temperature for 1h before samples were loaded. The 10 mg hepatic tissue was squeezed and lysed in

 

 

200 μl lysing buffer (50 mM NaCl, 1mM Na2 EDTA, 0.5% SDS, pH 8.3) for at least 30 min. For

 

electophoretic pattern of nucleic acids of tissue lysate, 20 μl of lysate hepatic cells was loaded in

 

well, 5 μl 6 × loading buffer was added on the lysing tissue.

 

 

 

105

 

 

DIETARY SUPPLEMENTATION AGAINST ALCOHOL TOXICITY

 

Electrophoresis was performed for 2 hours at 50 V in gel buffer (1 × TBE buffer). Gel was photographed

 

 

 

using a Polaroid camera while the DNA and RNA was visualized using a 312 nm UV transilluminator.

 

 

Nucleic acids extraction and molecular assessment for apoptosis

 

 

 

Nucleic acids extraction was based on salting out extraction method (Aljanabi and Martinez, 1997).

 

For apoptosis, the extracted DNA was gently resuspended with TE buffer supplemented with 5%

 

 

glycerol, gently pipetting, then the samples were mixed with 6 × loading buffer and loaded directly on

 

the gel (Hassab El-Nabi, 2004). The remained DNA was kept at –20oC for another loading. Apoptotic

 

 

 

bands appeared and located at 180, 360 and 540 bp.

 

 

Statistical analysis:

 

 

 

Data are presented as means ± s.d. in tables. Data were statistically analyzed by one-way analysis of

 

 

variance (Anova-Tukey test) using SPSS 10.1 software pakage. The P values < 0.05 were considered

 

 

 

significant.

 

 

RESULTS AND DISCUSSION

 

 

Effect of alcohol intake

 

 

 

The impact of prolonged alcohol consumption on the growth of experimental animals has been used

 

as a means to determine the overall toxicity of this compound (Wartburg and Popenberg, 1970). The

 

results of present study demonstrated significant decreases in body weight which reached its maximum

 

 

value (–20%) by the end of the 2nd week of alcohol intake (Table 1). The magnitude of depression in

 

 

 

body weight was attenuated thereafter by prolonged ethanol consumption (–15% after 8 weeks of

 

treatment). It is relevant in this respect to mention that chronic treatment of pregnant rats with

 

ethanol depressed feed and water consumption and weight gain (Abel, 1978). This depression in

 

weight gain of the alcohol-treated animals probably reflects the complication of the depressant

 

effects of ethanol on feed intake and impaired feed efficiency (Abel and Dintcheff, 1978).

 

The present data demonstrate that alcohol toxicity leads to not significant increase in the liver

 

weight and hepatosomatic index (Table 1). This alteration is accompanied with significant decreases

 

in total serum protein (Table 2). Such results lead to the suggestion that proteins are accumulated in

 

 

Table 1: Body weight, liver weight and hepatosomatic index during the trial

 

 

 

Control group Alcohol group Alcohol+antioxidant group

 

Rabbits, no. 5 5 5

 

 

Body weight (g): 2 week 1110±102b 890±143a 1010±82ab

 

 

 

4 week 1150±206 1046±177 1125±95

 

6 week 1270±186 1050±100 1150±98

 

 

8 week 1300±216b 1100±99a 1200±141ab

 

 

 

Liver weight (g): 2 week 59±14 60±12 60±14

 

4 week 60±15 64±14 61±10

 

6 week 61±15 68±16 61±11

 

8 week 61±16 72±16 62±14

 

 

Hepatosomatic index1 (%) 2 week 5.0±0.5 6.3±1.0 5.2±0.6

 

 

 

4 week 4.9±0.7 6.1±1.2 5.0±0.8

 

6 week 4.7±0.5 6.0±0.9 4.8±0.9

 

8 week 4.6±0.6 5.9±0.9 4.6±1.7

 

 

1 Hepatosomatic index: liver weight (g) / body weight (g) x 100). a, b: P<0.05

 

 

 

106

 

 

SHALAN et al.

 

 

 

the liver instead of being transported to blood, in response to toxic effects of alcohol. It is of

 

significant importance, in this context, to mention the results of Shalan (1995) showing that hepatic

 

protein content markedly increased after alcohol intoxication.

 

Moreover, ethanol was reported to decrease the number of hepatic macrotubules which are the key

 

promoting secretion and intracellular transport of proteins and accordingly protein retention occurs

 

accompanied by accumulation of lipids due to the increase in fatty acid binding protein leading to

 

 

fatty liver and hepatomegally (Pignon et al., 1987), whereas other authors attributed hepatomegally,

 

 

 

produced in response to chronic alcohol consumption, to the increase in the size of the hepatic cells

 

 

and not to the increase of hepatocyte number (Israel et al., 1979). Cunnane et al., (1985) showed that

 

 

 

increased liver weights resulted from hepatic triglyceride accumulation after chronic alcohol abuse.

 

The decreased serum protein content might be interpreted in the light of the fact that alcohol inhibits

 

 

secretion of the newly synthesized glycoprotein and albumin by hepatocytes (Lakshman et al.,

 

 

 

1989).

 

Serum glucose was increased significantly at 6 and 8 weeks of alcohol consumption compared with

 

 

normal controls (Table 2). It was shown that alcohol induced hyperglycemia (Forsander et al., 1958)

 

 

 

that resulted from release of glucose from hepatic glycogen stores (Ammon and Estler, 1968). At the

 

same time decreased peripheral utilization of glucose with alcohol intake helps in rising blood glucose

 

 

level (Lochner et al., 1967); this may be associated with alcohol effect that decreases blood insulin

 

 

 

and rises glucagon level (Bucher and Weir, 1976).

 

 

Table 2: Serum total protein and glucose concentration.

 

 

 

Control group Alcohol group Alcohol+antioxidant group

 

Rabbits, no. 5 5 5

 

Total protein (g/l): 2 week 59.2±5.2 53.8±5.1 54.6±5.7

 

 

4 week 62.2±8.9b 50.9±5.2a 57.2±5.3ab

 

6 week 60.0±7.7b 44.8±4.7a 53.4±5.8b

 

8 week 63.6±5.4b 43.0±4.5a 56.4±6.1b

 

 

 

Glucose (mmol/l): 2 week 4.92±0.51 5.28±0.54 5.02±0.52

 

4 week 4.67±0.48 5.50±0.60 4.87±0.54

 

 

6 week 4.66±0.47b 5.61±0.64a 4.78±0.49b

 

8 week 5.02±0.52b 6.77±1.04a 5.89±0.69ab

 

a, b: P<0.05

 

Table 3: Serum triglycerides and cholesterol concentration

 

 

 

Control group Alcohol group Alcohol+antioxidant group

 

Rabbits, no. 5 5 5

 

 

Triglycerides (mmol/l): 2 week 1.07±0.16b 1.58±0.17a 1.44±0.15a

 

4 week 1.11±0.18b 1.61±0.18a 1.45±0.14a

 

6 week 1.16±0.14b 1.66±0.23a 1.54±0.16a

 

8 week 1.21±0.19b 1.64±0.20a 1.43±0.16ab

 

 

 

Triglycerides (mmol/l): 2 week 2.58±0.30 3.03±0.42 2.70±0.29

 

 

4 week 2.48±0.32b 3.07±0.30a 2.62±0.36ab

 

 

 

6 week 2.41±0.44 2.77±0.39 2.59±0.31

 

8 week 2.45±0.73 2.70±0.46 2.53±0.55

 

 

a, b: P<0.05

 

 

 

107

 

 

DIETARY SUPPLEMENTATION AGAINST ALCOHOL TOXICITY

 

 

 

Ethanol induced significant increase in triglycerides concentrations at 2, 4, 6 and 8 weeks of treatment

 

compared with normal controls (Table 3). A significant increase in serum cholesterol content was

 

reported only at 4 weeks of treatment of rabbits with ethanol compared with normal controls (Table

 

 

3). Fatty liver is an important feature of alcohol abuse (Bernal et al., 1992). Alcohol decreased fatty

 

 

 

acids oxidation levels in the liver (Lieber, 1991) and that resulted in hepatic triglycerides accumulation

 

 

(Lamb et al., 1994). Baraona et al. (1973) indicated that rising serum triglycerides level related to

 

 

 

increased triglycerides synthesis resulted from increased fatty acids and alpha-glycerophosphate

 

availability during alcohol metabolism, and seemed that alcohol enhanced lipogenesis through

 

microsomes enhancement (Jenkins, 1984).

 

Results showed increased AST, ALT, ALP and LDH activities in response to alcohol administration

 

(Table 4). It was documented that alcohol causes modifications in the fluidity of membranes (Freund,

 

 

1979), permeability of these membranes (Ross, 1977), and their lipid composition (Hoek et al., 1988).

 

 

 

Therefore, alcohol may exert its effect through alteration of synthesis in the endoplasmic reticulum,

 

intracellular translocation and/or possibility of solubilization at the site of plasma membrane, hence

 

increasing the level of serum enzymes especially membrane-bound enzymes, like ALP, and cytosolic

 

enzymes, such as LDH and transaminases (ALT and AST). Elevated serum levels of ALP and LDH

 

 

were also observed by Yokoyame et al. (1993). Meanwhile, it was showed in previous studies that

 

 

 

chronic alcohol intoxication depresses hepatic enzyme activities, suggesting that elevated serum

 

enzyme activities might be induced as a result of enhanced release of hepatic enzymes into blood

 

stream due to liver cell injury (Singer and Kaplan, 1978).

 

Ethanol induced apoptotic DNA fragmentation of hepatocytes (Figure 1). Studies in mice and rats

 

revealed that both acute and chronic alcohol administration resulted in significant increases in

 

 

hepatocyte apoptosis (Goldin et al., 1993, Yacoub et al., 1995). Potential mechanisms of acute ethanolinduced

 

 

 

liver apoptosis include increased cytokine activity, Fas ligand (FasL) expression, and/or

 

 

Table 4: Serum ALT, AST, ALP and LDH activities (U/L)

 

 

 

Control group Alcohol group Alcohol+antioxidant group

 

Rabbits, no. 5 5 5

 

 

ALT activity (U/l): 2 week 62±9b 165±9a 62±7b

 

4 week 61±9b 170±10a 64±9b

 

6 week 61±6b 172±8a 65±8b

 

8 week 61±10b 170±6a 67±8b

 

AST activity (U/l): 2 week 183±20b 235±23a 187±16b

 

4 week 182±16b 233±20a 183±19b

 

6 week 181±20b 249±22a 182±16b

 

8 week 185±10b 243±17a 186±13b

 

ALP activity (U/l): 2 week 162±17b 200±24a 166±19b

 

4 week 163±11b 200±22a 164±13b

 

6 week 150±16b 202±19a 162±24b

 

8 week 162±13b 207±19a 165±13b

 

 

 

LDH activity (U/l): 2 week 5.12±0.63 6.76±0.95 6.11±0.58

 

 

4 week 5.18±0.56b 7.98±0.84a 6.05±0.64b

 

6 week 5.21±0.50b 9.17±0.94a 5.36±0.63b

 

8 week 5.24±0.b 10.96±0.94a 5.26±0.59b

 

a, b: P<0.05

 

 

 

108

 

 

SHALAN et al.

 

oxidative stress (Kurose et al., 1997, Neuman et al., 2001). Ethanol-induced liver apoptosis involves

 

the activation of cysteine proteases or caspases (Deaciuc et al., 1999, Zhou et al., 2001).

 

 

 

Endonucleases and DNA fragmentation factors are activated during apoptosis, resulting in degradation

 

 

of chromatin DNA into internucleosomal units (Cohen and Duke, 1984, Liu et al., 1997).

 

 

 

 

Effect of antioxidant treatment

 

 

 

Supplementations with vitamins (C, E), selenium and silymarin reduced the effect of alcohol intake

 

on body weight (Table 1). There were significant differences in serum total protein and glucose

 

concentrations between alcohol and alcohol + antioxidants groups at 6 weeks of treatment only

 

(Table 2). Ethanol + antioxidants group showed increased triglycerides content significantly at 2, 4

 

and 6 weeks of treatment compared with normal controls, however not significant differences was

 

reported at 8 weeks (Table 3). Supplementations with antioxidants significantly reduced the effect of

 

alcohol intake on serum ALT, AST, ALP and LDH activities (Table 4).

 

Overall results indicated the highly protective effects of vitamins (C, E), selenium and silymarin

 

supplements against alcohol intoxication. Tawfik (1998) reported that vitamin E protects

 

polyunsaturated fatty acids from oxygen effects, and inhibits lipid peroxidation enhanced by ethanol

 

 

(Situnayake et al., 1990) by acting as a free radical scavenger. Vitamin E stabilizes biomembranes and

 

 

 

prevents lysis of phospholipids (Koning and Drijver, 1979). It was shown that vitamin E prevents

 

 

alterations in ionic permeability of cellular membrane occurred after alcohol intake (Aono et al., 1978,

 

 

 

Littleton, 1980).

 

Vitamin C acts as a free radical scavenger and reduces alcohol capacity for interacting with critical

 

 

molecules (Davidson, 1998). Blankenship et al. (1997) indicated that vitamin C protects cells from

 

undergoing apoptosis. Upasani et al. (2001) showed that the preventive activity of vitamins C and E

 

 

 

may related to their antioxidant efficacy that inhibits lipid peroxidation.

 

 

Figure 1: Apoptotic DNA fragmentation in liver of rabbits treated with ethanol and the protective role of

 

 

 

vitamins (C, E), selenium and silymarin supplements. Lanes 1 and 2: ethanol treated with antioxidant

 

supplements for 8 weeks; Lanes 3, 4 and 5: ethanol only intake for 8 weeks; Lanes 6 and 7: controls; M: 1 kp

 

ladder.

 

 

109

 

 

DIETARY SUPPLEMENTATION AGAINST ALCOHOL TOXICITY

 

 

 

Silymarin is a natural mixture of antioxidants acting as free radical scavenger and preventing lipid

 

 

peroxidation (Soto et al., 1998). It was reported that silymarin improves liver function tests related to

 

hepatocellular necrosis and/or increases membrane permeability (Buzzelli et al., 1993). Ramadan et

 

al. (2002) reported that the protective effect of silymarin was attributed to its antioxidant and free

 

 

 

radical scavenging properties. It was suggested that silymarin modulates the cellular immunoresponse

 

 

and restores impaired liver function through its antioxidant capacity (Horvath et al., 2001). Feeding

 

 

 

of animals on silymarin-phospholipid complex normalized lipid metabolism and inhibited

 

atherosclerosis (Bialecka, 1997). The protective effect of silymarin may be attributed to its ability to

 

 

scavenge oxygen free radicals and to inhibite liver microsome lipid peroxidation (Mira et al., 1994).

 

 

 

It was recorded that alcohol intake may result in a decreased intake of other nutrients, maldigestion

 

and malnutrition (Lieber, 1988). Supplementation with antioxidants may repair the nutritional factors

 

that may be affected by ethanol toxicity. The combined antioxidant supplementation may alter ethanol

 

metabolism through their antioxidant capacities and thereby decreasing its toxic effects. Supplements

 

may exert their effects through rapid elimination of ethanol via bile or decreasing ethanol intestinal

 

absorption. Such suggestions need further studies to ensure the beneficial role of combined treatment,

 

through measuring blood ethanol concentrations and serum alcohol dehydrogenase concentrations.

 

In conclusion, the present study showed that treatment of alcoholic abused animals with vitamins (C

 

and E), selenium and silymarin supplements reduced toxic effects of ethanol.

 

 

REFERENCES

 

 

 

Abel E.L., 1978. Effects of ethanol on pregnant rats and their

 

 

offspring. Psychopharmacology, 57, 5-11.

 

 

 

Abel E.L., Dintcheff B.A., 1978. Effects of prenatal alcohol exposure

 

 

on growth and development in rats. J. Pharmacol. Exp. Therap.,

 

207, 916-921.

 

 

 

Aljanabi S.M., Martinez I., 1997. Universal and rapid saltextraction

 

of high quality genomic DNA for PCR-based

 

 

techniques. Nucl. Acids Res., 25, 4692-4693.

 

 

 

Ammon A.P., Estler C.J., 1968. Inhibition of ethanol induced

 

glycogenolysis in brain and liver by adrenergic beta –

 

 

blockade. J. Pharm. Pharmacol., 20, 164–165.

 

 

 

Aono K., Michio Y., Sosuke I., Kozo U.O., 1978. Radiation

 

 

protection of vitamin E. Okayama Igakkai. Zasshi., 90, 1297-

 

1308.

 

 

 

Apte U.M., McRee R., Ramaiah S.K., 2004. Hepatocyte proliferation

 

is the possible mechanism for the transient decrease in liver

 

 

injury during steatosis stage of alcoholic liver disease. Toxicol.

 

Pathol., 32, 567-576.

 

 

 

Baraona E., Pirda R.C., Lieber C.S., 1973. Pathogensis of

 

postprandial hyperlipemia in rats fed ethanol containing diets.

 

 

J. Clin. Inverst., 52, 296-303.

 

 

 

Bernal C.A., Vasquez Z.J.A., Adibi S., 1992. Liver triglyceride

 

concentration and body protein metabolism in ethanol treated

 

rats: Effect of energy and nutrient supplementation.

 

 

Gastroenterology, 103, 289-295.

 

 

 

Bialecka M., 1997. The effect of bioflavonoids and lecithin on the

 

 

course of experimental atherosclerosis in rabbits. Ann. Acad.

 

Med. Stetin., 43, 41-56.

 

 

 

Blankenship L.J., Caliste D.L., Wise J.P., Orenstein J.M., Dye L.E.,

 

Patierno S.R., 1997. Induction of apoptotic cell death by

 

particulate lead chromate: differential effects of vitamin C and E

 

 

on genotoxicity and survival. Toxicol. Appl. Pharmacol., 146,

 

 

 

270-280.

 

Bucher N.L.R., Weir G.C., 1976. Insulin, glucagone, liver

 

 

regeneration and DNA synthesis. Metabolism, 25, 1423-1425.

 

 

 

Buzzelli G., Mosarella S., Giusti A., Duchini A., Morena C.,

 

Lampertieo M., 1993. A pilot study on the liver: protective

 

effect of silybin-phosphatidyl choline complex (IDB 1016) in

 

 

chronic active hepatitis. Int. J. Clin. Pharmacol. Ther. Toxicol.,

 

31, 456-460.

 

 

 

Cohen J.J., Duke R.C., 1984. Glucocorticoid activation of a calciumdependent

 

endonuclease in thymocyte nuclei leads to cell death.

 

 

J. Immunol., 132, 38-42.

 

 

 

Cunnane S.C., Maku M.S., Horrobin D.F., 1985. Effect of ethanol on

 

liver triglycerides and fatty acid composition in the Golden

 

 

Syrian Hamster. Ann. Natr. Metab., 29, 246-252.

 

Davidson V., 1998. Vitamins and minerals. In: Davidson V.L.,

 

 

Sittman D.B.,(ed). 3rd edition biochemistry. Harwal

 

publishing, Baltimore, USA, 305-323.

 

 

 

Deaciuc I.V., Fortunato F., D’Souza N.B., Hill D.B., Schmidt J., Lee

 

E.Y., McClain C.J., 1999. Modulation of caspase-3 activity and

 

Fas ligand mRNA expression in rat liver cells in vivo by alcohol

 

 

and lipopolysaccharide. Alcohol Clin. Exp. Res., 23, 349-356.

 

 

 

Forsander O.A., Fartia K.O., Krusius F.E., 1958. Alcohol induced

 

 

hyperglycemia in man. Med. Exp. Fenn., 36, 1-8.

 

 

 

Freund G., 1979. Possible relationships of alcohol in membrane to

 

 

cancer. Cancer Res., 39, 2899-2901.

 

 

 

Goldin R.D., Hunt N.C., Clark J., Wickramasinghe S.N., 1993.

 

Apoptotic bodies in a murine model of alcoholic liver disease:

 

 

reversibility of ethanol-induced changes. J. Pathol., 171, 73-

 

 

 

76.

 

Hagymasi K., Koscsis I., Lugasi A., Fesher J., Blazovics A., 2002.

 

Extrahepatic biliary obstruction: Can silymarin protect liver

 

 

function. Phytother. Res., 16, 78-80.

 

 

 

Hassab El-Nabi S.E., 2004. Molecular and cytogenetic studies on

 

the antimutagenic potential of eugenol in human lymphocytes

 

 

culture treated with depakine and apetryl drugs. J. Egypt Ger.

 

Soc. Zool., 43, 171-196.

 

 

 

Hoek J.B., Taraschi T.F., and Robin A., 1988. Functional implications

 

of the interaction of ethanol with biologic membranes: Actions

 

 

of ethanol on hormonal single transduction systems. Seminars

 

in Liver Disease, 8, 36-46.

 

 

 

Horvath M.E., Gonzalez C.R., Blazovics A., Van der looij M., Barta

 

I., Muzes G., Gergely P., Feher J., 2001. Effect of silibinin and

 

 

110

 

 

SHALAN et al.

 

 

 

vitamin E on retardation of cellular immune response after partial

 

 

hepatoctomy. J. Ethanopharmacol., 77, 227-232.

 

 

 

Israel Y., Khanna J.M., Orrego H., Rachamin G., Wahid S., Britton

 

R., Macdonald A., Kalant H., 1979. Studies on metabolic

 

tolerance to alcohol, hepatomegally and alcoholic liver disease.

 

 

Drug Alcohol Depend., 4, 109-116.

 

Jenkins W., 1984. Liver disorders in alcoholism. In: Rosalki S.B.

 

 

 

 

(ed). Clinical biochemistry of alcoholism. Churchill

 

 

 

Livingstone press, Edinburgh, Germany. 262.

 

 

 

Konings A.W.T., Drijver E.B., 1979. Radiation effects in membranes.

 

 

I. Vit. E deficiency and lipid peroxidation. Rad. Res., 80, 494-

 

 

 

501.

 

Koyuturk M., Bolkent S., Ozdil S., Arbak S., Yanardag R., 2004.

 

The protective effect of vitamin C, vitamin E and selenium

 

combination therapy on ehtanol-induced duodenal mucosal

 

 

injury. Hum. Exp. Toxicol., 23, 391-398.

 

Kumar P., Clark M., 2002. Alcohol. In: Kumar and Clark clinical

 

medicine., W.B. Saunders, London, UK, 250-251.

 

 

 

Kurose I., Higuchi H., Miura S., Saito H., Watanabe N., Hokari R.,

 

Hirokawa M., Takaishi M., Zeki S., Nakamura T., Ebinuma H.,

 

Kato S., Ishii H., 1997. Oxidative stress-mediated apoptosis of

 

 

hepatocytes exposed to acute ethanol intoxication. Hepatology,

 

 

 

25, 368-378.

 

Lakshman M.R., Chirtel S.J., Chambers L.C., Cambell B.S., 1989.

 

Hepatic synthesis of apoproteins of very low density and high

 

density lipoproteins in perfused rat liver: influence of chronic

 

 

heavy and moderate doses of ethanol. Alcohol. Clin. Exp. Res.,

 

13, 554-559.

 

 

 

Lamb R.G.G., Koch J.C., Snyder J.W., Hudand S.M., Bush S.R., 1994.

 

 

A model of ethanol dependent liver injury. Hepatology, 19,

 

174-182.

 

 

 

Lieber C.S., 1988. The influence of alcohol on nutrional status.

 

 

Nutr. Rev., 46, 241-254.

 

 

 

Lieber C.S., 1990. Mechanism of ethanol induced hepatic injury.

 

 

Pharmacol. Therap., 41, 1-41.

 

Lieber C.S., 1991. Alcohol and the liver.In: Palmer T.N. (ed). The

 

 

 

 

molecular pathology of alcoholism. Oxford University Press,

 

 

 

New York, USA, 60-129.

 

 

 

Littleton J.M., 1980. The effects of alcohol on the cell membrane: A

 

 

possible basis for tolerance and dependence. In: Richter D. (ed).

 

 

 

 

Addiction and brain damage. Oxford University Press, New

 

 

 

York, USA, 46-76.

 

 

 

Liu X., Zou H., Slaughter C., Wang X., 1997. DFF, a heterodimeric

 

protein that functions downstream of caspase-3 to trigger DNA

 

 

fragmentation during apoptosis. Cell, 89, 175-184.

 

 

 

Lochner A., Wulff J., Madison L.L., 1967. Alcohol and peripheral

 

 

utilization of glucose. Metabolism, 16, 1-12.

 

 

 

Marino M.D., Aksenov M.Y., Kelly S.J., 2004. Vitamin E protects

 

against alcohol-induced cell loss and oxidative stress in the

 

 

neonatal rat hippocampus. Int. J. Dev. Neurosci., 22, 363-377.

 

 

 

Martin J.V., Nolan B., Wagner G.C., Fisher H., 2004. Effect of dietary

 

caffeine and alcohol on liver carbohydrate and fat metabolism

 

 

in rats. Med. Sci. Monit., 10, 455-461.

 

 

 

Mira I., Silva M., and Mauso C.F., 1994. Scavenging of reactive

 

 

oxygen species by silibinin dihemisuccinate. Biochem.

 

Pharmacol., 48, 733-739.

 

 

 

Neuman M.G., Grenner D.A., Rehermann B., Taieb J., Chollet-Martin

 

S., Chhard M., Garaus, J.J., Poynard T., Katz G.G., Cameron R.G.,

 

Shear N.H., Gao B., Takamatsu M., Yamauchi M., Ohata M., Saito

 

S., Maeyama S., Uchikoshi T., Toda G., Kumagi T., Akbar S.M.,

 

Abe M., Michitaka K., Horiike N., Onji M., 2001. Mechanisms

 

 

of alcoholic liver disease: cytokines. Alcohol Clin. Exp. Res.,

 

25(Suppl), 251-253.

 

 

 

Pignon J.P., Bailey N.C., Baraona E., Lieber C.S., 1987. Fatty acidbinding

 

protein: a major contibutor to ethanol induced increase

 

 

in liver cytosolic proteins in the rat. Hepatology, 7, 865-871.

 

 

 

Ramadan L.A., Roushdy H.M., Abu Senna G.M., Amin N.E., El-

 

Deshw O.A., 2002. Radioprotective effect of silymarin against

 

 

radiation induced hepatotoxicity. Pharmacol. Res., 45, 447-

 

454.

 

 

 

Ross D.H., 1977. Adaptive changes in Ca++ membrane interactions

 

 

following chronic ethanol exposure. Adv. Exp. Med. Biol., 85,

 

 

 

459-471.

 

 

Ruf J.C., 2004. Alcohol, wine and platelet function. Biol. Res., 37,

 

 

 

209-215.

 

Saito Y., Yoshida Y., Akazawa T., Takahashi K., Niki E., 2003. Cell

 

death caused by selenium deficiency and protective effect of

 

 

antioxidants. J. Biol. Chem., 278, 39428-39434.

 

 

 

Shalan A.G., 1995. Physiological and biochemical studies of effects

 

 

of ethyl alcohol on liver and testes of the growing rat. M. Sc.

 

 

 

 

Thesis, Menoufia University, Egypt.

 

 

 

Shalan M.G., 1996. Biochemical studies of effects of alcohol

 

consumption on fat and carbohydrate metabolism in rats fed

 

 

different levels of proteins. M. Sc. Thesis, Faculty of Science,

 

 

 

 

Menoufia University, Egypt.

 

 

 

Sherlock S., Dooley J., 2002. Alcohol and the liver. In: Diseases of

 

 

 

 

the liver and biliary system. Blackwell Science publishing Ltd.,

 

 

 

Eleventh ed., Milan, Italy, 381-398.

 

 

 

Singer J.S., Kaplan M.M., 1978. Ethanol depresses rat liver gammaglutamyl

 

 

transpeptidase. Gastroenterology, 74, A 1095.

 

 

 

Situnayake R.D., Crump B.J. and Thurnhan D.I., 1990. Lipid

 

peroxidation and hepatic antioxidants in alcoholic liver

 

 

disease. Gut, 31, 1311.

 

 

 

Soto C.P., Perez B.L., Favari L.P., and Reyes J.L., 1998. Prevention

 

of alloxan-induced diabetes mellitus in the rat by silymarin.

 

 

Comp. Biochem. Physiol. Pharmacol. Toxicol. Endocrinol., 119,

 

 

 

125-129.

 

Tawfik S.S.M., 1998. Radio-protective role of antioxidant vitamins

 

 

in irradiated albino-mice. M. Sc. Thesis, The Institute of

 

 

 

 

Environmental studies and Research, Ain Shams University,

 

Egypt.

 

 

 

Upasani C.D., Khera A., Balaraman R., 2001. Effect of lead with

 

vitamins E, C, or spirulina on malondialdehyde conjugated

 

 

dines and hydroperoxides in rats. Indian J. Exp. Biol., 39, 70-

 

 

 

74.

 

Wartburg J.P., Popenberg J., 1970. Biochemical and enzymatic

 

 

changes induced by chronic ethanol intake. In: International

 

 

 

 

Encyclopedia of Pharmacology and Therapeutics. Vol. 11,

 

 

 

Pergamon Press, Oxford, USA, 301-343.

 

 

 

Yacoub L.K., Fogt F., Nanji A.A., 1995. Apoptosis and Bcl-2

 

expression in experimental alcoholic liver disease in the rat.

 

 

Alcohol Clin. Exp. Res., 19, 854-859.

 

 

 

Yokoyame H., Ishii H., Nagata S., Kato S., Kamegaya K., Tsuchiya

 

M., 1993. Experimental hepatitis induced by ethanol after

 

 

immunization with acetaldehyde adducts. Hepatology, 17, 14-

 

 

 

 

19.

 

 

 

Zhou Z., Sun X., Kang Y.J., 2001. Ethanol induced apoptosis in

 

mouse liver: Fas and cytochrome c-mediated caspase-3

 

 

activation pathway. Am. J. Pathol., 159, 329-338

 

 

 

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