
enhanced, compared to control group, in chronically
ethanol-treated rats (59, 55, and 71 %, respectively).
When animals were treated with H. scoparia extracts
(DE and ME), the activities of aminotransferases were
practically restored and reached the normal values of
control rats. Particularly, the beneficial effect of the
ME treatment on the biochemical parameters of liver
injury was highly significant (P<0.001).
Evaluation of lipid peroxidation and antioxidant
enzymes (CAT, SOD, and GPx)
Liver TBARS levels, a lipid peroxidation marker,
increased significantly after alcohol treatment by a
factor of 2.8 compared with controls (Fig. 2a). This
effect was completely inhibited in the presence of H.
scoparia extracts, especially in ME extract enriched in
flavonoids (Table 1).
To evaluate the ability of ME to prevent ROS-
induced oxidative stress, SOD, CAT, and GPx activi-
ties were meas ured in liver homogenates. Figure 2b–d
shows that rats treated with ethanol exhibited a
marked decrease in the activities of SOD, CAT, and
GPx. Treatment with H. scoparia extracts (HE, DE,
and ME) could not repair the reduction in SOD and
CAT activities, while GPx activity was restored after
H. scoparia methanolic extract treatment.
Histological examination showed a protective effect
of ME
The liver sections of ethanol-intoxicated rats proved
massive ballooning degeneration and cytoplasmic
vacuolization (Fig. 3b) compared to a normal histolo-
gy in control livers (Fig. 3a). The hepatocellular dam-
age was slightly reduced in ethanol-fed rats treated
with HE or DE (Fig. 3c, d). Nevertheless, the histo-
logical architecture of liver sections of the rats treated
with methanolic extract demonstrated prominent re-
covery in the form of maintained hepatic histoarchi-
tecture (Fig. 3e), such as reduced cytoplasmic
vacuolization and ballooning degeneration.
Glycogen synthase kinase
Immunoblot analysis was used to examine the conse-
quences of chronic ethanol consumption on total
GSK-3β in liver h omogenate. The data presented in
Fig. 4 show that GSK-3β was overexpressed after
ethanol treatment. However, ethanol was not able to
induce such effect in rats r eceiving H. scoparia
extracts (HE, DE, and ME). We suggest that ethanol
activates GSK-3β, which was demonstrated using the
anti-phospho Ser
9
GSK-3β anti body (Fig. 4).
Discussion
The liver is the major site of xenobiotic metabolism
and excretion. This organ accounts for 90 % of alcohol
metabolism. Indeed, it is the most adversely affected
organ after an excessive consumption of alcohol. The
alcohol absorbed from the intestinal tract gains access
first and foremost to the liver, resulting in a variety of
liver ailments. Thus, liver diseases remain one of the
serious health problems due to alcohol abuse [6].
In the present study, the hepatoprote ctive effect of
hexane, dichlorom ethane, and methanolic extracts of
H. scoparia leaves was evaluated in alcoholic rat
model a nd find out the therapeutically efficacious
extract. An attempt was made to find out the correlation
Table 2 The total phenolic content and antioxidant activities in
vitro (2,2-Diphenyl-1-picrylhydrazyl and β-carotene bleaching
method)
Extract TPC (mg of gallic
acid equivalents/g)
HE (hexane extract) 9.76±0.030
DE (dichloromethane extract) 34.75±0.085
ME (methanol extract) 58.82±0.082
Total polyphenolic content in the extracts of H. scoparia leaves
expressed in terms of gallic acid
Table 3 The total phenolic content and antioxidant activities in
vitro (2,2-Diphenyl-1-picrylhydrazyl and β-carotene bleaching
method)
Extract IC
50
(μg/ml) AAC
HE (hexan extract) 23.6 485
DE (dicloromethan extract) 6.3 606
ME (methanolic extract) 2.6 666
α-Tocopherol (antioxidant reference) 5.5 805
Ascorbic acid (antioxidant reference) 3.5 –
BHT (antioxidant reference) – 753
DPPH-scavenging activity of plant extracts (IC
50
) and antioxi-
dant assay using the β-carotene bleaching method (AAC anti-
oxidant activity coefficient)
232 E. Bourogaa et al.