Iraqi J Pharm Sci , Vol.18 (Suppl.), 2009                                                  Benfotiamine in liver injury 

                                  

 

 74 

Study of the Protective Effects of Benfotiamine Against CCl4-Induced 

Hepatotoxicity in Rats 
Tavga A. Aziz 

*
,   Zheen A. Ahmed 

**
,   Kasim M. Juma'a 

*** 

Munaf H. Abdulrazzaq 
****

    and   Saad A. Hussain
****,1

 
*
Department of Pharmacology, College of Pharmacy, University of Sulaimaniya, Kurdistan, Iraq 

**
 Department of Pharmacology, College of medicine, University of Sulaimaniya, Kurdistan, Iraq 

***
Department of Pharmacy, Baquba General Hospital, Diyala, Iraq 

****
Department of Pharmacology and Toxicology, College of Pharmacy,University of Baghdad, Baghdad, Iraq 

 

Abstract 
Liver is considered as the first target for the toxic effects of toxins and other xenobiotics, and this 

can be attributed to its role as a site which receive all absorbed xenobiotics from the gastrointestinal 

tract and its role as a major site for biotransformation of xenobiotics. The present study was designed to 

evaluate the possible hepatoprotective effect of benfotiamine against CCl4-induced hepatotoxicity in 

rats. The study was conducted on 48 male albino rats; the animals were allocated into 8 groups (6 rats 

in each group) and treated as follow: 4 groups treated with oral doses of either normal saline, 

benfotiamine (100 mg/kg), thiamine (100 mg/kg), N-acetylcystein (400 mg/kg) only without induction 

of hepatic damage. The other 4 groups were treated as indicated previously with induction of hepatic 

damage with CCl4; at the end of treatment period, rats were scarified, blood samples obtained and 

livers excised for the assessment of the oxidative stress parameters (MDA and GSH), cholesterol and 

triglycerides levels. Additionally, serum levels of total bilirubin, albumin, total protein and the 

activities of ALT, AST and ALP enzymes were evaluated before and after treatment with 

benfotiamine. Tissue sections were prepared for evaluation of histopathological changes. The results 

indicated that benfotiamine has the ability to protect hepatic tissue against the toxicity induced by CCl4, 

revealed through reduction of serum levels of TSB and liver enzymes, decrease in the hepatic tissue 

MDA levels and elevation of GSH there. Histological evaluation of tissue sections prepared for this 

purpose confirmed the previous finding. In conclusion, benfotiamine is capable to protect liver tissue 

against CCl4-induced toxicity in rats more than thiamine.  

Key words: Benfotiamine, CCl4, Hepatotoxicity 

 الخالصة
ٚعحبر انكبد انٓدف األٔل نهحأذٛر انطاو نهًركبات انًخحهفة نكَّٕ انًٕقع األٔل انذ٘ ٚطحهى شًٛعع انًركبعات انحعٙ ٚعحى ايحااععٓا  عٍ 

طرٚععا اناُععاه انٓ ععًٛة اكععا ة نكَٕععّ يركععس انحلععٕ ت األٚ ععٛة نٓععذِ انًركبععاتس جععى جاععًٛى ْععذِ اندعاضععة نحاٛععٛى انلًاٚععة انًحٕقعععة نًععا ه 

شعراا جبعٛح ثٛعد جعى جاطعًٛٓا انعٗ  7٤ٛايٍٛ كد انحطًى انًطحلدخ بربا ٙ كهٕعٚد انكربٌٕ  ٙ انصعرااٌس جشرٚعث اندعاضعة  هعٗ انبُفٕج

يلهععٕل يهلععٙ ٕاثععد يععٍ انًركبععات انحانٛععة  نبصععرع  ععٍ طرٚععا انفععى  يُٓععا شععرااٌ نكععم يصًٕ ععةال جععى  عع ز جعبعععة٦ذًاَٛععة يصًٕ ععات  

يهغى/كغعىس جيعا انًصًٕ عات األعبععة األفعرٖ  حًعث  7١١يهغى/كغعى، جضعٛحٛم ضطعحٍٛٛ  ٠١١ٍٛ يهغى/كغعى، ذٛعاي ٠١١يحٕازٌ، بُفٕجٛايٍٛ 

يعانصحٓا بُفص انطرٚاة انًذكٕعه آَفا يع اضحلداخ جهف كبد٘ بٕاضطة عبا ٙ كهٕعٚد انكربٌٕ، ٔ ٙ َٓاٚة  حره انع ز جى قحم انلٕٛاَعات 

ًانَٕعدا٘ اندٚٓاٚعدال، انكٕنٛطعحرٔل ٔانمعلٕو  عٙ نانحأكطعد  انكهٕجعاذٌٕٛ ٔا نهلإل  هٗ جكبا ْا ٔ ُٛات يٍ  يٓعاس جعى قٛعاش يععاٚٛر  عرط

ال  عٙ ياعم  AST ،ALT ،ALPف عة َطٛس انكبد باألكا ة انٗ قٛاش يطحٕٖ يا ه انافراء، انبعرٔجٍٛ ٔ عانٛعة األَسًٚعات انكبدٚعة  

س جى جل ٛر يااطع َطٛصٛة نهكبعد ٔيحابععة انحغٛٛعرات فرٖاندو ٔيااعَة يطحٕٖ ْذِ انًعاٚٛر قبم ٔبعد انع ز بانبُفٕجٛايٍٛ ٔانًركبات األ

انلاعهة بٕاضطة انًصٓرس جذبحث َحائس اندعاضة يادعه انبُفٕجٛايٍٛ  هٗ ثًاٚة َطٛس انكبد كد انحطًى انًطحلدخ بٕاضعطة عبعا ٙ كهٕعٚعد 

انٗ ففح يطحٕٚات انكٕنٛطحرٔل ٔانمعلٕو انكربٌٕ يٍ ف ل ففح يطحٕٚات يا ه انافراء ٔ عانٛة األَسًٚات انكبدٚة  ٙ اندو، جكا ة 

 ٔانًانَٕدا٘ اندٚٓاٚد  ٙ َطٛس انكبد يع ع ع يطحٕٖ انكهٕجاذٌٕٛ ُْاك، كًا جكدت َحائس انفلص انُطٛصٙ يااكر آَفعاس ًٚكعٍ األضعحُحاز بعأٌ

 رااٌ اكرر يٍ انرٛايٍٛس نهبُفٕجٛايٍٛ انًادعه  هٗ ٔقاٚة َطٛس انكبد كد ان رع انًطحلدخ بٕاضطة عبا ٙ كهٕعٚد انكربٌٕ  ٙ انص

 

Introduction 
The liver plays a central role in 

carbohydrate, protein and fat metabolism and 

allows the detoxification of various 

xenobiotics. Additionally, it regulates the 

synthesis and secretion of bile. 
(1)

 Toxic injury 

occurs in the liver more often than other 

organs, because all ingested substances that are 

absorbed are first presented to the liver and 

that the liver is responsible for the metabolism 

and elimination of many substances.
(1)

Many 

xenobiotics such as acetaminophen, CCl4 ,and 

yellow phosphorus produce liver damage in a 

predictable and dose-dependent manner; the 

most frequent mechanism of hepatocellular 

injury involves production of injurious 

metabolites by the cytochrome P450 system. 
(2,3)

 Preventive care can significantly reduce the 

progression of liver disease. 
(4)

  

 
1 
Corresponding author E-mail :  saad_alzaidi@yahoo.com 

Received   : 3/1/2009 

Accepted   : 6/6/2009  

mailto:saad_alzaidi@yahoo.com


Iraqi J Pharm Sci , Vol.18 (Suppl.), 2009                                                  Benfotiamine in liver injury 

                                  

 

 7٤ 

One of the drugs that used for this purpose 

is N-Acetylcysteine (NAC) which has been 

used clinically for the treatment of 

Acetaminophen poisoning 
(5)

. Oral 

supplementation with (NAC) provides an 

alternate means of boosting intracellular 

glutathione via elevated intracellular cysteine,
 

and this can scavenge peroxynitrite and 

hydroxyl radicals as well as convert hydrogen 

peroxide to water.
 (6)

 Benfotiamine (S-benzoyl 

thiamine-O-monophosphate) has been shown 

to reduce the formation of advanced glycation 

end products (AGE) byactivating 

transketolase. 
(7, 8)

 Benfotiamine has been 

noted to possess clinical efficacy in the 

treatment of diabetic cardiomyopathy, 
(9)

 

diabetic nephropathy 
(10)

 and diabetic 

neuropathy. 
(11, 12)

 Moreover, benfotiamine has 

been shown to reduce the oxidative stress 

through a mechanism unrelated to AGE 

formation. 
(13)

 Activation of Akt (protein 

kinase B) has been demonstrated to stimulate 

eNOS activity, increase the bioavailability of 

NO and reduce the generation of ROS. 
(14)

 

Benfotiamine has been reported to improve the 

function of endothelium by activating Akt and 

subsequently stimulating eNOS and inhibiting 

the generation of ROS.
 (15,16,17)

 Benfotiamine is 

thought to act by at least three different 

mechanisms. First, activation of the 

hexosamine pathway with subsequent decrease 

in the accumulation of deleterious glucose 

metabolites seems to be involved, second, 

normalization of PKC activity along with 

prevention of nuclear factor kappa B (NF-κB) 

activation has been found in retinas, and third, 

correction of imbalances in the polyol pathway 

by decreasing aldose reductase activity, 

sorbitol concentrations and intracellular 

glucose levels seems to play a role. 
(7)

 

Absorption of benfotiamine was better than 

thiamine itself, and levels of thiamine and 

thiamine pyrophosphate (TPP) remain higher 

for longer period of time. 
(18)

 Absorption of 

thiamine in the form of benfotiamine is found 

to be five times greater than the absorption of 

the conventional thiamine supplements, and 

because of greatest intracellular access of 

benfotiamine, its tissue availability and effects 

are more impressive, especially in the brain 

and muscles. 
(19)

 The present study was 

designed to evaluate the possible protective 

effect of benfotiamine against CCL4-induced 

hepatotoxicity in rats. 

 

Materials and Methods  
Forty eight Sprague-Dawley rats of both 

sexes weighing 180-220 g were obtained from 

the animal house in the College of Pharmacy, 

University of Baghdad and used in the study. 

The animals were housed in the animal house 

of Collage of Pharmacy, University of 

Sulaimaniya under conditions of controlled 

temperature and allowed free access to water 

and food. The study protocol was approved by 

the Committee for Medical Research, College 

of Medicine, University of Sulaimaniya.   The 

animals were allocated into eight groups, each 

contain 6 rats and treated as follow: Group I, 

received single oral daily dose of normal saline 

for 7 days. The group served as control; group 

II received single oral daily dose of normal 

saline for 7 days, at day 8 the animals received 

single dose of CCl4 (2 ml of a mixture of 1:1 

v/v in a corn oil /kg/day) orally by oral needle 

to induce liver damage. 
(20)

 The animals were 

sacrificed 24 hr after CCl4 administration; 
 (21)

 

group III received single oral daily dose of 

thiamine (70 mg/kg/day) for 7 days; group IV 

received single daily oral dose of thiamine (70 

mg/kg/day) started 7 days prior to treatment 

with CCl4 at day 7; group V received single 

oral daily dose of benfotiamine (70 mg/kg/day) 

for 7 days; group VI received single daily oral 

dose of benfotiamine (70 mg/kg/day) started 7 

days prior to treatment with CCl4 at day 7;  

Group VII received single oral daily dose of 

N-acetylcysteine (400 mg/kg/day) for 7 days; 

group VIII received single daily oral dose of 

N-acetylcysteine (400 mg/kg/day) started 7 

days prior to treatment with CCl4 at day 7. All 

animals were sacrificed on the day 8. After 

killing the animals by over dose of thiopental 

(Panpharma S.A. France) (100mg/kg), livers 

were obtained and utilized for preparation of 

tissue homogenate. The organ was quickly 

existed and placed in a chilled phosphate 

buffer solution (pH 7.4), blotted with filter 

paper, and 10% homogenate was prepared in 

the same solution utilizing metal head tissue 

homogenizer at 4
o
C. All preparations were 

frozen (-18 
0
C) unless worked immediately. 

After killing the animals, blood was collected 

by intracardiac puncture. The clot was 

dispersed with glass rod and then centrifuged 

for 15-20 minutes at 2000 rpm and the 

supernatant was used for the estimation of 

ALT and AST, 
(22)

 ALP, 
(23)

 albumin and total 

protein 
(24)

 as parameters of liver function tests 

and total serum bilirubin 
(25)

 as excretory 

function test. Samples of liver tissue 

homogenates were used for determination the 

malondialdehyde (MDA), 
(26)

 reduced 

glutathione (GSH), 
(27)

 cholesterols and 

triglycerides levels. 
(28,29)

 Tissue sections were 

prepared for histological examination 

according to the method of Bauer, 
 (15)

 using 

paraffin sections technique. The significance 

of differences between the mean values was 

calculated using unpaired Student's t -test. P-



Iraqi J Pharm Sci , Vol.18 (Suppl.), 2009                                                  Benfotiamine in liver injury 

                                  

 

 74 

Values less than 0.05 were considered 

significant for all data shown in the study. 

 

Results 
Table 1 showed that serum activities of 

alanine-aminotransferase (ALT) and aspartate 

aminotransferase (AST) were significantly 

elevated in CCl4-intoxicated animals (847.5% 

and 2381.6% respectively) compared to 

control group (P>0.05). Pre-treatment of rats 

with single oral doses of Thiamine, 

Benfotiamine or N-Acetylcysteine for 7 days 

prior to intoxication with CCl4 showed a 

marked decline in the serum ALT and AST 

activities (36.6%, 32.8% and 88.6% 

respectively for ALT and 39.75%, 77.97% and 

94.2% respectively for AST) compared to 

CCl4-treated animals (P>0.05). Serum alkaline 

phosphatase (ALP) activity was significantly 

elevated in CCl4-treated animals (53.4%) 

compared to control group (P>0.05, table 1), 

and pre-treatment with single oral doses of 

Thiamine, Benfotiamine or N-Acetylcysteine 

for 7 days, prior to induction of liver toxicity 

with CCl4, showed a marked decline in the 

level of serum ALP activity (17%, 37.9% and 

32.2% respectively) compared to CCl4-treated 

animals.

 

Table 1. Effects of Benfotiamine on serum levels of ALT, AST and ALP activities in experimental 

animal model of CCl4-induced liver toxicity. 

Type of Treatment Serum ALT U/L Serum AST U/L Serum ALP U/L 

Saline treated only 8.0 ± 1.06  6.0 ± 0.6 79.3 ± 18.3 

Saline + CCl4    75.8 ± 11.5 * 
a
    148.9 ± 17.5 * 

a
 121.7 ±26.6*

 a
 

Thiamine only 10.2 ± 1.5 
b
     6.5 ± 0.71 

b
 90.5 ± 19.1*

 b
 

Thiamine + CCl4   48.0 ± 9.5 * 
c
      89.7 ± 16.1 * 

c
 101.0 ±20.2* 

b
 

Benfotiamine only 9.6 ± 1.2 
b
   7.1 ± 0.9 

b
 80.9 ± 8.1 

c
 

Benfotiamine + CCl4  50.9 ± 8.2 * 
c
     32.8 ± 6.1 * 

d
 75.5 ± 10.6 

c
 

N-Acetylcysteine only 9.0 ± 1.2 
b
 7.2 ± 0.9 

e
 72.0 ± 9.0 

c
 

N-Acetylcysteine + CCl4 8.6 ± 1.4 
b
    8.6 ± 1.2 * 

e
 82.5 ± 10.2

 c
 

Each value represents mean ± SD; * significantly different compared to saline only treated group 

(P<0.05); values with non-identical superscripts (a,b,c,d,e) are considered significantly different within 

the same parameter (P<0.05). 

 

Table 2 showed that animals intoxicated 

with CCl4 presented with a highly significant 

increase in MDA (95.6%) and decrease in 

GSH (72.2%) contents of liver tissue 

homogenate  compared to control (saline 

treated) group (P>0.05, table 2). Pre-treatment 

of rats orally with single doses of Thiamine, 

Benfotiamine or N-Acetylcysteine for 7-days 

before induction of hepatotoxicity with orally 

administered CCl4 resulted in 32%, 31.1% and 

51% decrease in hepatic MDA contents 

respectively, and significant increase in GSH 

(340%, 237% and 246% respectively) 

compared to CCl4- treated only animals  

(P>0.05, table 2). 

 

Table 2. Effect of Benfotiamine on liver tissue malondialdehyde (MDA) and reduced glutathione 

(GSH) levels in experimental animal model of CCl4-induced liver toxicity 

Type of Treatment 

 

Liver tissue GSH 

µmol/g tissue 

Liver tissue MDA 

nmol/g tissue 

Saline 27.0 ± 6.75 230.0 ± 46.0 

Saline + CCl4 7.5 ± 0.81*
a
 450.0 ± 48.0*

a
 

Thiamine 21.0 ± 4.2 
b
 246.0 ± 36.0 

b
 

Thiamine + CCl4 33.0 ± 6.0*
c
 306.0 ± 14.0*

c
 

Benfotiamine 25.5 ± 5.2 
b
 218.0 ± 35.0 

d
 

Benfotiamine + CCl4 25.3 ± 2.6 
b
 310.0 ± 22.0*

c
 

N-Acetylcysteine 31.2 ± 4.5 
c
 221.0 ± 26.0 

d
 

N-Acetylcysteine + CCl4 26.0 ± 5.3 
b
 220.0 ± 18.0 

d
 

Each value represents mean ± SD; * significantly different compared to saline only treated group 

(P<0.05); values with non-identical superscripts (a,b,c,d) are considered significantly different within 

the same parameter (P<0.05). 

 

 



Iraqi J Pharm Sci , Vol.18 (Suppl.), 2009                                                  Benfotiamine in liver injury 

                                  

 

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In table 3, total serum bilirubin (TSB) 

levels were significantly elevated in CCl4-

treated animals (85.7%) compared to control 

group (P>0.05, table 3), and pre-treatment 

with single oral doses of Thiamine, 

Benfotiamine or N-Acetylcysteine for 7 days, 

prior to induction of liver toxicity with CCl4, 

showed a marked decline in the level of serum 

TSB (33.8%, 41.5% and 30.7% respectively) 

compared to CCl4-treated animals. Serum 

albumin and total protein were shown to be 

significantly decreased in CCl4-treated animals 

(31.8%) compared to control group (P<0.05), 

while total protein levels showed a non 

significant differences in CCl4-treated animals 

(1.3%) compared to control group (Table 3). 

Pre-treatment of rats with single oral doses of 

Thiamine, Benfotiamine or N-Acetylcysteine 

for 7 days, prior to intoxication with CCl4, 

showed increase in  the level of serum albumin 

(45.4%, 36.36% and 13.6% respectively) 

compared to CCl4-treated animals (P>0.05, 

table 3), while total protein levels were not 

significantly changed. However, rats treated 

with single oral dose of Thiamine or 

Benfotiamine for 7 days showed non-

significant differences on both serum albumin 

and total protein (P < 0.05) compared to 

control group. Meanwhile, single oral dose of 

N-Acetylcysteine for 7 days showed a 

significant increase in both serum albumin and 

total protein levels (P > 0.05) compared to 

control group (Table 3; figure 4).Serum 

Triglyceride and Cholesterol levels in liver 

tissue homogenate were shown to be 

significantly increased in CCl4-intoxicated 

animals (333% and 212.5% respectively) 

compared to control group (P> 0.05, table 3). 

However, Pre-treatment of rats with single oral 

doses of Thiamine, Benfotiamine or N-

Acetylcysteine for 7 days, prior to induction of 

hepatotoxicity with CCl4, showed a marked 

decline in the levels of triglycerides (20%, 

57% and 58% respectively) and 

cholesterol(24.5%, 44.9% and 42.2% 

respectively) compared to CCl4-treated 

animals (P>0.05, table 3 ). Histological 

examination of liver tissues of rats exposed to 

toxic dose of CCl4 showed a marked hepatic 

damage revealed as zonal necrosis, extensive 

diffuse vacuolar degeneration of the 

hepatocytes, together with ballooning and fatty 

changes, dilatation and congestion of the 

central vein; the later showed focal 

hemorrhage with variable degree of 

inflammatory cell reaction were seen also 

(figure 1). Treatment of rats with 

Benfotiamine, thiamine or N-acetylcysteine 

before induction of toxicity clearly 

demonstrated protective effects against CCl4-

induced toxicity, where Benfotiamine showed 

the greatest level of protection compared to 

that produced by N-acetylcysteine and 

thiamine respectively (figures 2, 3 and 4). 

 

Table 3. Effect of Benfotiamine on serum levels of total serum bilirubin, albumin, total protein 

and Liver tissue Triglycerides and Cholesterol in experimental animal model of CCl4-induced 

liver toxicity 

Type of Treatment 

 

Total Serum 

Bilirubin 

mg/dl 

 

Serum 

albumin 

mg/dl 

 

Liver tissue 

Triglycerides 

µmol/g tissue 

 

Liver tissue 

Cholesterol 

µmol/g tissue 

 

Serum total 

protein 

mg/dl 

 

Saline treated only 0.35 ± 0.01 2.9 ± 0.08 12.0 ± 2.4 8.8 ± 0.3 6.17 ± 0.4 

Saline + CCl4 0.65 ± 0.14* 
a
 2.2 ± 0.1* 

a
  52.0 ± 7.2 *

a
  18.7 ± 2.1*

a
 6.3 ± 0.6 

a
 

Thiamine only 0.31 ± 0.02 
b
 3.2 ± 0.2 *

b
 11.5 ± 2.1

b
 9.3 ± 1.8

b
 5.9 ± 0.38

 a
 

Thiamine + CCl4 0.43 ± 0.1* 
c
 3.2 ± 0.1* 

b
 41.5 ± 3.6 *

a
 14.1 ± 3.6*

c
 5.8 ± 0.3

 a
 

Benfotiamine only 0.29 ± 0.03 
b
 2.9 ± 0.1 

b
 10.2 ± 1.3 

b
   8.5 ± 0.8 

b
 5.3 ± 0.7

 a
 

Benfotiamine + CCl4 0.38 ± 0.04 
d
 3.0 ± 0.1 

b
 22.3 ± 3.4 *

c
   10.3 ± 0.9 *

d
 5.9 ± 0.5

 a
 

N-Acetylcysteine only  0.39±0.02 
d
 4.5 ± 0.4 * 

c
 9.3 ± 1.6

 b
 8.3 ± 0.6 

b
 7.2 ± 1.2* 

b
 

N-Acetylcysteine + CCl4   0.45 ± 0.02*
c
 2.5 ± 0.2 

b
 21.8 ± 2.6 *

c
   10.8 ± 0.5 *

d
 6.0 ± 1.1

a
 

Each value represents mean ± SD; * significantly different compared to saline only treated group 

(P<0.05); values with non-identical superscripts (a,b,c,d) are considered significantly different within 

the same parameter (P<0.05). 

 
 
 
 
 
 
 



Iraqi J Pharm Sci , Vol.18 (Suppl.), 2009                                                  Benfotiamine in liver injury 

                                  

 

 0٠ 

 
Figure 1. Section from liver tissue after 

CCl4 intoxication. (X400, H and E stain) 

 

 
Figure 2. Section of liver tissue after 

intoxication with CCl4 and protection with 

orally administered N-acetylcysteine. 

(X400; H and E stain) 

 

 
 

Figure 3. Section from liver tissue after 

CCl4 intoxication and protection with orally 

administered thiamine (X 10; H and E 

stain) 

 

 
Figure 4. Section of liver tissue after 

intoxication with CCl4 and protection with 

orally administered Benfotiamine. (X400; H 

and E stain) 

 
Discussion 

In animals acutely exposed to CCl4 orally, 

the liver appears to be the primary target 

organ.
(21)

 Numerous studies showed that 

metabolism of (CCl4) is required for its 

toxicity;
 (30)

 it is known to be rapidly 

transformed by cytochrome P450-2E1 of the 

hepatocyte endoplasmic reticulum to CCl3
•
 

which is converted into CClOO
• 

in the 

presence of oxygen.
(31)

 Peroxidative 

degradation of membrane lipids of 

endoplasmic reticulum rich in polyunsaturated 

fatty acids leads to the formation of lipid 

peroxides, these in turn give products like 

MDA that cause damage to the membrane
 (32)

 

and alter cellular function.
(33)

 The reduction in 

GSH is due to consumption for conjugation of 

metabolites, and then redox potential of the 

tissue will be impaired.
(34)

 GSH depletion also 

results in lipid peroxidation and impaired 

antioxidant enzyme activities.
(35)

 The data 

presented in this study clearly demonstrated 

the state of oxidative stress induced in hepatic 

tissues by CCl4 treatment manifested by 

elevation of MDA content in tissue 

homogenate, which is associated with 

depletion of GSH content, these results are 

compatible with those obtained by others.
(36)

 

Administration of Benfotiamine results in 

increased intracellular thiamine diphosphate 

levels, a cofactor of transketolase enzyme; 

activation of this enzyme by thiamine may 

reduce superoxide overproduction through 

directing advanced glycation and lipoxidation 

end products substrates to the pentose 

phosphate pathway.
(7,8)

 The data presented in 

this work showed a significant decline in 

MDA level and increase in GSH level in 

animals treated with Benfotiamine and 

thiamine prior to CCl4 administration, this 

might be attributed to the up-regulation of 

transketolase but could be a positive side effect 

of Benfotiamine, which show an intrinsic 

antioxidative activity by itself.
(37)

 It is 

important to mention that the effect of 

Benfotiamine was much better than thiamine 

concerning GSH levels. Lipid peroxidation and 

oxidative stress were attenuated by NAC 

administration to the rats prior to CCl4, which 

may be due to enhancement of intracellular 

GSH biosynthesis. Lipid peroxidation has been 

proposed to disrupt cellular membrane, 

resulting in loss of membrane integrity,
 (38)

 and 

may lead to leakage of ALT and AST and 

increasing their activities in the plasma. The 

plasma transaminases (ALT and AST) are 

known to be increased significantly in rats 

after exposure to toxic doses of CCl4.
(39)

 The 

data presented in table 1, clearly support the 



Iraqi J Pharm Sci , Vol.18 (Suppl.), 2009                                                  Benfotiamine in liver injury 

                                  

 

 05 

above explanation and seems to be consistent 

with those obtained by others. 
(40)

 The sharp 

elevation in serum activities of the enzymes 

that localized in bile ducts is a useful 

biochemical index for bile duct damage, 

particularly alkaline phosphatase (ALP). In the 

present work, serum activity of alkaline 

phosphatase that is present in the lining 

membrane of the hepatocytes was also 

increased in the CCl4-treated rats compared to 

normal control animals, and these results are 

consistent with that reported by other 

investigators.
(41)

 The major finding of the 

present study is that treatment of animals with 

Benfotiamine prior to CCl4 elicits beneficial 

effects on the structure and functions of the 

liver; regarding the enzymatic activities, there 

is a significant decrease in AST level 

compared with that intoxicated with CCl4, the 

reduction was three times more than that 

obtained through the use of thiamine prior to 

CCl4 administration, but, still the protection 

was less than that observed in the animal group 

treated with NAC prior to CCl4. The data also 

revealed attenuation of ALT activity induced 

by CCl4 due to the use of Benfotiamine, which 

was approximately the same as that noted with 

thiamine; however, the protection was much 

less than that observed due to the use of NAC. 

Concerning serum ALP, the serum activity 

level of this enzyme almost normalized by 

Benfotiamine, again the protection was better 

than thiamine even it precede that of NAC. 

The data presented in this study regarding the 

effect of NAC was consistent with those 

observed by others.
(38)

 The suggested 

mechanisms behind these effects of 

Benfotiamine may be due to indirect action of 

the lipophylic pro-drug through decreasing 

lipid peroxidation, which is the main factor 

affecting membrane permeability and integrity, 

and consequently preventing further leakage of 

cytosolic enzymes;
 (38)

 another possible 

explanation may be related to its negative 

action on protein kinase C activities.
(7,8)

 It is 

important to mention that the impact of 

Benfotiamine was better than that of both 

thiamine and NAC when each one 

administered alone prior to CCl4 in this 

respect. In conclusion, Benfotiamine protects 

hepatic tissue of rats against CCl4-induced 

damage. 

 

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