_____________________________________________________________________________________________________ [600881]

_____________________________________________________________________________________________________

*Corresponding author: E
–
mail:
[anonimizat]
,
[anonimizat]
;

International Journal of Biochemistry Research

& Review

12
(
2
):
1
–
9
, 20
16
, Article no.IJBCRR.26029

ISSN: 2231
–
086X, NLM ID: 101654445

SCIENCEDOMAIN

international

www.sciencedomain.org

LIGHT and FGF
–
BP are Upregulated in Chronic Liver
Injury

Elsayed Gomaa Elsayed Elsakka
1*

1
Department of
Biochemistry,
Faculty
of
Pharmacy
, Alazhar University, Cairo, Egypt.

Author
’
s

contribution

The sole author
designed, analyzed and interpreted and prepared the manuscript.

Article Information

DOI: 10.9734/
IJBC
RR/201
6
/
26029

Editor(s):

(1)
Chunying Li, Department of Biochemistry and Molecular Biology Wayne State University School of Medicine,

Detroit, USA
.

Reviewers:

(1)
Kelvin K
.

Juma, Kenyatta University, Kenya
.

(2)

Anonymous, University of Alberta, Canada
.

Complete
Peer review History:

http://sciencedomain.org/review
–
history/14478

Received
30
th

March
20
16

Accepted
26
th

April

20
16

Published
5
th

May

20
16

ABSTRACT

Aims:
The aim of this study is to
explore the expression pattern of LIGHT and FGF
–
BP at the
different stages of liver injury including acute injury, fibrosis and cirrhosis
.

Study
Design
:

Controlled experiment
.

Place and Duration of Study:

Department of Biochemistry and Department of Pharmacology and
Toxicology, Faculty of pharmacy (boys) Al
–
Azhar University, between February 2015 and June
2015.

Methodology:

Four Sprague
–
Dawley rat groups were used for the exp
eriment.

Control group:

9
rats received corn oil; acute toxicity group: 10 rats were injected 50% CC
l
4
in corn oil (4
ml/kg/IP/single dose); 6

weeks group:

1
2

rats were injected with 50% CCl
4
in corn oil (4
ml/kg/IP/twice weekly/6

weeks); 11

weeks group: 1
0
rats were injected with 50% CCl
4

in corn oil (4
ml/kg/IP/twice weekly/6 w
eeks) followed by 5 weeks of CCl
4
treatment with the half previous dose
.

On the day after the last dose, rats were anesthetized with diethyl ether and blood samples were
collected
for measurement of blood chemistry. The animals were then euthanized, and tissue
samples from the livers harvested and divided into 2 parts; one part was processed for standard
histology and immunofluorescence techniques and the other was homogenized for o
xidative status
assessment.

Results:

LIGHT and FGF
–
BP were shown to upregulate in chronic liver injury; while LIGHT was
Original Research Article

Elsakka; IJBCRR,
12
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2
):
1
–
9
, 20
16
; Article no.
IJB
C
RR
.
26029

2

shown to upregulate in cirrhosis stage only, FGF
–
BP was shown to upregulate at fibrosis and
cirrhosis stages. LIGHT was shown to expres
s at the cytoplasm of the cells at cirrhotic nodule
border. FGF
–
BP localization was shown to occur mainly at the extracellular matrix.

Conclusion:

LIGHT and FGF
–
BP are of critical importance in pathogenesis of chronic liver injury
so they may be used as a
target for chronic liver injury therapy and/or candidate marker for
diagnosis and/or prognosis of chronic liver injury.

Keywords:

LIGHT
;

FGF
–
BP
;

fibrosis
;

cirrhosis
.

1.
INTRODUCTION

L
iver plays an unlimited role in the preservation
and body homeostasis regulation. It is included in
most biochemical pathways to growth, protection
against disease, nutrient fund, energy facility

and reproduction
[
1
]
.

Hepatotoxicity can be
generated by certain common causes including
therapeutic agents, natural chemicals, laboratory
and manufacturing agents and herbal therapies
[
2
]
.

Carbon tetra chloride (
CCl
4
) is one of the most
communal models for inducing hepatotoxicity. It

is transformed into

a toxic CCl
3
–

radical by
hepatic cytochrome P4502E1 (CYP2E1).Thus; it
brings an acute Centro
–
lobular necrosis which
starts a wound healing response (fibrosis)
[
3
]
.
However, Contact to these chemicals in humans
is rare and generally occurs in the manufacturing
during fa
brication and in places where these
chemicals is usually used
[
4
]
.

LIGHT (lymphotoxin
–
like, exhibits induci
ble
expression, and competes with HSV glycoprotein
D (gD) for HVEM, a receptor expressed by T
lymphocytes) It is a cytokine in the TNF
superfamily that is involved in innate and
adaptive immune responses as well as in
regulation of cell survival and prolif
eration

[
5
]
.
LIGHT was demonstrated to have a crucial role
in fibrous tissue precipitation and fibrosis
cascades in different tissues
[
5
–
8
]
.

Fibroblast growth factor binding proteins (FGF
–
BPs) comprise a family of three secreted proteins
that are of vital import
ance for fibroblast growth
factors (FGFs) action
[
9
]
. They are reported
to be
upregulated in tissue healing and repair
specifically those that include some sorts of
fibrosis cascades
[
10
,
11
]
.

The aim of this study was to explore the
expression pattern of LIGHT and FGF
–
BP at
different stages of liver injury. This was verified
by investigation of
the localization of these
proteins in different liver injury stages.

2.
METHODOLOGY

2.1
Animal
Model

Adult male Sprague
–
Dawley rats weighing 250
–
300 g were used in the current study. The
animals were obtained from the breeding colony
maintained at the animal house of the Nile
Company for pharmaceuticals, Cairo, Egypt.
They were housed in the animal facili
ty of Faculty
of Pharmacy, Al
–
Azhar University in 20 X 18 X 25
cm plastic cages with stainless steel wire lids and
mesh floor with 5 animals per cage. They were
kept at 23±1°C, at 55% relative humidity, with
12:12
–
h light: dark cycle, and maintained on a
s
tandard rodent chow (El
–
Nasr Company, Abou
–
Zaabal, Cairo, Egypt), diets and water
ad libitum
.
Animals were randomly divided into four groups:
control group: 9 rats: receiving corn oil

(2

ml/kg/twice weekly/IP) ; acute toxicity group
:
10 ra
ts were injected 50% CCl
4

in corn oil

(4 ml/kg/IP/single dose)

[
12
]
; 6

weeks group:

1
2

rats wer
e injected with 50% CCl
4

in corn oil (4
ml/kg/IP/twice weekly/6

weeks) and finally 11

weeks group: 10
rats were injected with 50%
CCl
4

in corn oil (4 ml/kg/IP/twice weekly/6 weeks)
followed by 5

weeks of CCl
4

treatment with the
half previous dose. On the d
ay after the last
dose, rats were anesthetized with diethyl ether
and blood samples were collected for
measurement of blood chemistry. The animals
were then euthanized, and tissue samples from
the livers were harvested and divided into 2
parts; one part wa
s processed by standard
histology and immunofluorescence techniques
and the other was homogenized in 0.15 M KCl
for MDA and SOD assay. All animal procedures
were performed in accordance with the
international guide for the care and use of
laboratory animal
s
[
13
]

2.2 Antibodies and chemicals

Rabbit polyclonal LIGHT and FGF
–
BP antibodies
were purchased from Santa Cruz Biotechnology
(CA, USA). Cy3
–
conjugated goat anti
–
rabbit
antibody was purchased from Jackson

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Immunoresearch (PA, USA). 4, 6
–

Diamid
ino
–
2
–

phenyl indole (DAPI). CCl
4

was purchased
from
Sigma
–
Aldrich (MO, USA).

2.3
Biochemical
Analysis

Serum enzymatic activities of transaminases
(ALT and AST) were estimated by kinetic method
according to the method of international
federation of clinical chemistry (IFCC)
(14;15)
.
Alkaline phosphatase (ALP) activity was assay
according to the method of IFCC

[
16
–
18
]

and
serum albumin concentration according to the
method described by Gendl
er
[
19
]
. The liver
homogenate was used for the determination of
the oxidative stress parameters.

The level of
thiobarbituric acid
–
reactive substances
(measured as malondialdehyde (MDA) was
determined as described by Mihara and
Uchiyama
[
20
]
. The activity of SOD was
determined using the method described by
Marklund

[
21
]
.

2.4
Immunofluorescence
Analysis

Slides containing liver tissues sections were
handled according to method described by
Abdel
–
Bakky and his associates
[
22
]
. The
primary LIGHT and FGF
–
BP antibodies were
diluted in blocking solution in the suitable dilution
(1:400) an
d left overnight in 4°C. Secondary
antibody (cyanine red conjugated) diluted in the
blocking solution was incubated for 30 min and
the nuclei were counterstained using DAPI.
Finally, all slides were mounted with the
fluoromount solution, covered by coverin
g slips,
and allowed to stand for detection by
immunofluorescence microscope (Leica DM
5500B).

2.5 Statistical
Analysis

Data were presented as the mean ±SE.
Statistical analysis was performed using one
–
way
analysis of variance (ANOVA) followed by Tukey
–
K
ramer as a post hoc test, according to the
number of groups. The 0.05 level of probability
was used as the criterion of significance using
GraphPad Prism software version 5 (GraphPad
Software Inc, CA, USA).

3.
RESULTS

3.1

Liver Functions and Oxidative
Status
Assessment

To assess liver functions; we have performed
liver function tests including serum
transaminases, ALP, and albumin. The oxidative
status of liver tissues was determined through
MD
A and SOD assay as indicated in

Table 1.

As indicated in Table 1; Administration of I.P.
CCl
4

significantly elevated serum transaminases
activities compared to control whatever the
duration of administration.

As shown in Table 1; ALP activity showed
significant elevation only in chronic
administ
ration of
CCl
4

with m
inimal change in
single dose CCl
4

treatment compared to control
group.

Serum albumin concentration showed significant
decrease only with 11 weeks CC
l
4

treatment
compared to control group as shown in Table 1.

As shown in Table 1; the

tissue MDA content
was significantly increased on CC
l
4

treated
groups whatever the duration of administration
compared to control group.

Table 1 shows that CCl
4

treated groups exhibited
significant decrease in SOD activities compared
to control group.

3.2
Expression and Localization of Light

Fig. 1
shows that LIGHT protein showed well
defined expression only liver tissue that showed
well defined cirrhosis; while there no any definite
protein expression in normal liver tissues, acute
toxicity model and
chronic models that didn’t
show cirrhosis. It was also noted that the
expression was located mainly in the cirrhotic
nodule borders; area that showed high density of
fibroblasts while areas that showed parenchymal
cells as interior of the cirrhotic nodule
didn’t show
any expression. It was noted that the expression
in
creased by about 38% 11

weeks CCl
4

treated
group compared to control group while there
weren’t significant expression change in other
groups as indicated in
Graph

1
.

3.3

Expression and Localization of
FGF
–
BP

Fig
.

2

and
Graph
2

show that FGF
–
BP showed
minimal expression in normal liver tissues and
al
so in acute toxicity model. The expression was
increased on conversion of toxicity from acute to
chronic with maximal expression in 6 and 11
weeks groups. It also can be noted that FGFBP
was located mainly in the extracellular matrix of
necrotic area in in
jured tissue.

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As indicated in
Graph

2
; quantitative analysis of
immunofluorescence staining expressed as
fluorescence intensity revealed that there was no
significant change in the expression between
control and singly treated gr
oup. On the other
hand; the expression increased by about
58%
and 32% in 6 and 11

weeks CCl
4

treated groups
respectively compared to control group.

3.4
Histopathological Findings

Effects of CCl
4

treatment on histopathological

findings of liver tissue are represented in

Table

and Fig. 3. The control group showed
normal hepatic architecture. On contrast, CCl
4

treated group
s showed pathological findings
changes according to the duration of treatment.

Table
1
.

Liver functions and oxidative status assessment tests

ALT(IU/L)

AST(IU/L)

ALP(IU/L)

Albumin

(g/dl)

MDA(nmol/g
tissue)

SOD(U/mg
tissue)

Control

46.6±2.85

54.1±2.65

312±21.9

3.81±0.08

4.73±0.318

526±25.2

CCl
4

single
I.P. dose

294±11.9
(a)(b)

419±11.0
(a)

395±11.1
(b)

3.8±0.06
(b)

23.7±2.35
(a)(b)

308±8.05
(a)(b)

CCl
4

6 weeks
I.P.

349±13.4
(a)

436±15.4
(a)

570±24.5
(a)

3.39±0.13

31.9±1.44
(a)

223±14
(a)

CCl
4
11
weeks I.P.

298±8.48
(a)

368±13.4
(a)(b)

790±24.9
(a)(b)

2.69±0.07
(a)

38.1±1.74
(a)

171±6.57
(a)

Data are expressed as mean ± SEM

(a) Significantly different from control group

(b) Significantly different from 6 weeks CC
l
4

group using one
–
way ANOVA followed by Tukey
–
Kramer test for multiple
comparison test at P
≤0.05

Fig.
1
.

Effect of CCl
4

administration on hepatic expression of LIGHT protein

Immunofluorescence staining of liver sections of CC
l
4

treated rats showing minimal LIGHT expression in control
group, single I.P. treated group and s
ix weeks CCl
4
treated group. The expression was increased significantly in
11 weeks group.it was noted that the expression located mainly in the cytoplasm of ce
lls located in fibrous septa
(white arrow) with minimal expression in the interior of

cirrhotic nodule (blue arrows)

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5

LIGHT
control
single I.P. CCL4
6 weeks I.P.CCL4
11 weeks I.P. CCL4.
0
50
100
150
a
Flourescence intensity
% of control
b
b
b

Graph
1
.

Effect of CCl
4

administration on
hepatic expression of LIGHT protein

Fluorescence intensity was obtained from 5 fields of
each section (minimally 2 rats of each group) using
ImageJ software.

Data are expressed as mean ± SEM

(a) Significantly different from control group

(b) Signific
antly different from 6 weeks CCl
4

group
one
–
way ANOVA followed by Tukey
–
Kramer test for
multiple comparison test at P
≤0.05

FGFBP
control
single I.P. CCL4
6 weeks I.P.CCL4
11 weeks I.P. CCL4.
0
50
100
150
200
a
a,b
b
Flourescence intensity
% of control

Graph
2
.

Effect of CCl
4
administration on
FGF
–
BP

Fluorescence intensity was obtained from 5 fields
of
each section (minimally 2 rats of each group) using
ImageJ software.

Data are expressed as mean ± SEM

(a) Significantly different from control group

(b) Signific
antly different from 6 weeks CCl
4

group
one
–
way ANOVA followed by Tukey
–
Kramer test for
mult
iple comparison test at P
≤0.05

Fig
.

2
.

Effect

of CCl
4

administration on hepatic expression and localizaion of FGF
–
BP

Single I.P. dose treated group showed average
portal tract (PT)
(yellow arrow) without interface
activity or underlying fibrosis. On the other hand;
it showed spotty necrosis (black arrow), and mild
micro
–
vesicular steatosis and dilated central vein.
Six weeks treatment caused expanded PT with
fibrous septa extending f
rom PT to PT (yellow
arrows), marked micro
–

and macro
Table
2
.

Histopathological findings of the studied groups

Control

CV

0

Steatosis

0

Hepatocyte

0

Spotty necrosis

0

Interface activity

0

PT

0

Fibrosis

0

Central vein (CV):

0: within normal +: dilated ++: markedly dilated

Steatosis, Spotty

0: no +: mild

Hepatocytes:

0: within normal

Portal tract (PT):

0: within normal +: expanded
Fibrosis:

0: no fibrosis

+: fibrosis confined to enlarged portal zones

++: fibrosis of peri

+++
: architectural distortion (septal or bridging fibrosis) without obvious cirrhosis

++++: probable or definite cirrhosis

Fig
.

3
: Histopathological mi
crograph of liver samples of CCl
Control
: liver tissue showing average PT (yellow arrows), average central vein (red arrow), and average
hepatocytes arranged in cords.
Single I.P.
necrosis (black arrow), and mild micro
–
with underlying fibrosis and fibrous septa extending from PT to PT (yellow arrows), marked micro
vesicular steatosis.
11 weeks I.P. CC
l
nodular formation, marked micro
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Single I.P. dose treated group showed average
(yellow arrow) without interface
activity or underlying fibrosis. On the other hand;
it showed spotty necrosis (black arrow), and mild
vesicular steatosis and dilated central vein.
Six weeks treatment caused expanded PT with
rom PT to PT (yellow
and macro
–
vesicular

steatosis. The hepatocytes exhibited some
aspects of single cell necrosis with dilated central
veins and mild interface activity.

Eleven weeks treatment showed markedly
ex
panded PT (yellow arrows) with definite
inflammatory infiltrate, complete nodular
formation (definite cirrhosis) and marked

Histopathological findings of the studied groups

Single I.P.
CCl
4

CCl
4

6 weeks

CCl
+

+

++
+

+++

+++
++

+

+

++

0

+

0

+

++
0

+

++
0

++

++++
Central vein (CV):

0: within normal +: dilated ++: markedly dilated

Steatosis, Spotty
necrosis, Interface activity:

0: no +: mild

++: moderate to marked

Hepatocytes:

0: within normal
+: single cell necrosis ++: confluent or diffuse necrosis
Portal tract (PT):

0: within normal +: expanded

++: expanded with inflammatory infiltrate
0: no fibrosis

+: fibrosis confined to enlarged portal zones

++: fibrosis of peri
–
portal or portal
–
portal septa with intact architecture

: architectural distortion (septal or bridging fibrosis) without obvious cirrhosis
++++: probable or definite cirrhosis

crograph of liver samples of CCl
4

treated
groups ×235 using H and
E stain

: liver tissue showing average PT (yellow arrows), average central vein (red arrow), and average
Single I.P.
CCl
4

dose
: liver tissue showing average PT (yellow arrow), spotty
–
vesi
cular steatosis.
6 weeks I.P. CCl
4
: liver tissue showing expanded PT
with underlying fibrosis and fibrous septa extending from PT to PT (yellow arrows), marked micro
l
4
: liver tissue showing markedly expanded P
T (yellow arrows), complete
nodular formation, marked micro
–

and macro
–
vesicular steatosis

; Article no.
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26029

steatosis. The hepatocytes exhibited some
aspects of single cell necrosis with dilated central
Eleven weeks treatment showed markedly
panded PT (yellow arrows) with definite
inflammatory infiltrate, complete nodular

formation (definite cirrhosis) and marked
4

11 weeks

++

+++

++

++

++++

+: single cell necrosis ++: confluent or diffuse necrosis

++: expanded with inflammatory infiltrate

: architectural distortion (septal or bridging fibrosis) without obvious cirrhosis

groups ×235 using H and
: liver tissue showing average PT (yellow arrows), average central vein (red arrow), and average
: liver tissue showing average PT (yellow arrow), spotty
: liver tissue showing expanded PT
with underlying fibrosis and fibrous septa extending from PT to PT (yellow arrows), marked micro
–

and macro
–
T (yellow arrows), complete

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7

micro
–

and macro
–
vesicular steatosis. The
hepatocytes
exhibited single cell necrosis with
mild spotty necrosis. Finally; the central veins
were
markedly dilated with marked interface
activity.

4.
DISCUSSION

Administration of CCl
4

either a single I.P. dose or
chronic I.P. doses for 6 or 11 weeks significantly
elevated serum transaminases enzymes
activities suggesting hepatocellular damage.
These results agree
d with previous reports that
CCl
4

significantly increases serum
transamina
ses
[
23
–
25
]
. This CCl
4
induced hepatic
damage was reported to be due to oxidative
stress
[
26
,
27
]
.

ALP activity after singl
e I.P. CCl
4

treatment
shows non
–
significant increase from control
group suggesting minimal biliary injury detected
upon that model. This result opposes what was
reported by Kālu’s team

[
24
]
. This difference
might be due difference in animal strain or route
of administration that has not been mentioned.
On the other hand; ALP elevation in chronic
models including 6 weeks, 11 weeks groups
showed signi
ficant increase compared to control
which is less than 3 times as control; a result that
might suggest hepatotoxicity and mild biliary
toxicity. This matches with that previously
reported by Posen and Doherty

[
28
]
.

Prolonged
CCl
4

for 11 weeks has showed significant
elevation in ALP
activity compared to 6 weeks
CCl
4

treatment suggesting that chronic
hepatocellular intoxication might lead to biliary
injury.

Serum albumin concentration showed only
significant decrease from control

at 11 weeks
CCl
4

treatment, a group that showed cirrhotic
nodule formation. This result agreed with
previous reports that indicated the decrease in
serum albumin concentration upon decreased
liver functioning tissue mass

[
29
]
.

Meanwhile; Free radicals production and
oxidative stress are main player
s in liver injury
especially CCl
4

induced liver injury
[
26
,
27
]
. In
our
study; administration of CCl
4

significantly
elevated tissue content of MDA and reduced
serum SOD activity. These results agreed with
the previous reports that demonstrated the great
role of oxidative stress in CC
l
4

induced liver
injury
[
30
–
32
]
.

Supporting the measured biochemical data: Our
findings demonstra
ted that single I.P. dose of
CCl
4

resulted in marked spotty necrosis and
confluent hepatocytes necrosis. On the other
hand

prolonged administration of CCl
4

to 6
weeks
produced fibrosis of peri
–
portal or portal
–
portal septa with intact
architecture while
prolonged CCl
4

to 11 weeks produced definite
cirrhosis with clear cirrhotic nodule. This is
similar to what has been reported that
histopathologica
l changes occur accord
ing to
CCl
4

administrat
ion period. A single dose of CCl
4

leads to centrizonal necrosis and steatosis
[
33
]
,
while continued administration leads to hepatic
fibrosis, cirrhosis, and HCC
[
34
]
.

Concerning inflammatory cytokines; it was shown
that LIGHT protein expression showed significant
u
pregulation in 11 weeks group only. This might
suggest that LIGHT was greatly involved in
critical cellular processes that control cirrhosis
progression. This was quite similar to previous
reports that have indicated the importance of
LIGHT in fibrous tiss
ue precipitation and fibrosis
cascades in different tissues

[
5
–
8
,
35
]

and might
be an attractive marker and a target for fibrosis
therapies relevant to humans

[
36
]
.

Concerning FGF
–
BP; our results demonstrated
that FGF
–
BP is upregulated significantly in
chronic liver injury g
roups compared to control
group; groups with fibrosis and cirrhosis pattern
that have been shown to involve cellular
processes of classic wound healing and repair of
other tissues

[
37
]
. These finding agreed with
what has been reported that FGF
–
BP is of critical
importance in tissues that involve some sorts of
tissue healing or repair
[
10
,
11
]
.

5.
CONCLUSION

In conclusion, our findings indicate, for the first
time, that LIGHT and FGF
–
BP are
upregulated in
chronic not acute liver injury specifically in stages
of fibroblast activity. LIGHT is upregulated only in
cirrhotic stages while stages before did not. FGF
–
BP is upregulated in chronic liver injury stages
involving fibrosis and cirrhosis. T
his may be a
guide for usage of LIGHT and/or FGF
–
BP as a
candidate marker for diagnosis and/or prognosis
of chronic liver diseases and their usage as a
target for drug therapy in treatment of liver
diseases
.

COMPETING INTERESTS

Author has declared that

no competing interests
exist.

Elsakka; IJBCRR,
12
(
2
):
1
–
9
, 20
16
; Article no.
IJB
C
RR
.
26029

8

REFERENCES

1.

Sharma A, Chakraborti
KK
, Handa
SS
.
Antihepatotoxic
activity of some indian
herbal formulations as compared to
silymarin
. Fitoterapia
.

1991;62:229
–
35.

2.

Ostapowicz G, Fontana
RJ
, Schiodt
FV
,
Larson A, Davern
TJ
, Han
SH
,
et a
l.
Results
of a prospective study of acute
liver failure at 17 tertiary care centers in
the
United States. Ann Intern Med
.

2002;137(12):947
–
54.

3.

Domenicali M, Caraceni P, Giannone F,
Baldassarre M, Lucchetti G, Quarta C,
et
al.

A
novel model o
f
Ccl4
–
induced cirrhosis
with ascites in the mouse
. J Hepatol
.

2009;51(6):991
–
9.

4.

Mormone E, George J, Nieto N. Molecular
pathogenesis of hepatic fibrosis and
current therapeutic approaches
. Chem Biol
Interact
. 2011
;193(3):225
–
31.

5.

Granger
SW,
Rickert S. Lig
ht
–
Hvem
signaling and the regulation of
T
cell
–
mediated immunity.

Cytokine Growth
Factor Rev
.

2003;14(3
–
4):289
–
96.

6.

Doherty
TA
, Soroosh P, Khorram N,
Fukuyama S, Rosenthal P, Cho
JY
,
et al
.
The
tumor necrosis factor family member
light is a target for
asthmatic airway
remodeling
. Nature Medicine
.

2011;17(5):

596
–
603.

7.

Yamamoto T. The
bleomycin
–
induced
scleroderma model:

What
have we learned
for scleroderma pathogenesis?

Arch
Dermatol Res
.

2006;297(8):333
–
44.

8.

Herro R, Antunes
RS
, Aguilera
AR
,
Tamada K, Cr
oft M. The
tumor necrosis
factor superfamily molecule light promotes
keratinocyte activity and skin fibrosis.

J
Invest Dermatol
.

2015;135(8):2109
–
18.

9.

Abuharbeid S, Czubayko F, Aigner A. The
fibroblast growth factor
–
binding protein
Fgf
–
Bp. Int J Biochem Cel
l Biol
.

2006;38(9):

1463
–
8.

10.

Liu
XH
, Aigner A, Wellstein A, Ray
PE
. Up
–
regulation of a fibroblast growth factor
binding protein in children with renal
diseases.

Kidney Int
.
2001;59(5):1717
–
28.

11.

Kurtz A, Aigner A, Cabal
–
Manzano
RH
,
Butler
RE
, Hood
DR
, Sessions
RB
,
et al
.
Differential
regulation of a fibroblast growth
factor
–
binding protein during skin
carcinogenesis and wound healing
.
Neoplasia
.

2004;6(5):595
–
602.

12.

Fleurentin J, Hoefler C, Lexa A, Mortier F,
Pelt
JM
. Hepatoprotective
properties of
crepis rueppellii and anisotes trisulcus
:
Two
traditional medicinal plants of
Yemen.
J Ethnopharmacol
.

1986;16(1):105
–
11.

13.

Institute
of
Laboratory Animal Resources
(Us), Use
of
Laboratory Animals, National
Institutes
of
Health (Us). Guide
for t
he
Care
and
U
se
of
Laboratory Animals.
National Academies; 1985.

14.

Bergmeyer
HU
, Horder M, Rej R.
International Federation
of
Clinical
Chemistry (
IFCC
) Scientific Committee,
Analytical Section: Approved Recommen
–
dation (1985)
on
Ifcc Methods
for the
Measurement
of
Catalytic Concentration
of
Enzymes. Part 2. Ifcc Method
for
Aspartate
Aminotransferase (L
–
Aspartate: 2
–
Oxoglutarate Aminotransferase, Ec
2.6.1.1). J Clin Chem Clin Biochem
.

1986;24(7):497
–
510.

15.

Bergmeyer
HU
, Horder M, Rej R.
International Federation
of
Clin
ical
Chemistry (
IFCC
) Scientific Committee,
Analytical Section: Approved Recommen
–
dation (1985)
on
Ifcc Methods
for the
Measurement
of
Catalytic Concentration
of
Enzymes. Part 3. Ifcc Method
for
Alanine
Aminotransferase (L
–
Alanine: 2
–
Oxoglutarate Aminotran
sferase, Ec
2.6.1.2). J Clin Chem Clin Biochem
.

1986;24(7):481
–
95.

16.

Tietz
NW
, Rinker
AD
, Shaw
LM
.
International Federation
of
Clinical
Chemistry.
IFCC methods for the
measurement of catalytic concentration of
enzymes
. Part 5. Ifcc Method
for
Alkaline
Phosphatase (Orthophosphoric
–
Monoester
Phosphohydrolase, Alkaline Optimum, Ec
3.1.3.1). Ifcc Document Stage 2, Draft 1,
1983
–
03 With
a
View
to an
Ifcc
Recommendation. Clin Chim Acta
.
1983
;135(3):339f
–
67f.

17.

Tietz
NW
, Rinker
AD
, Shaw
LM
.
IFCC
methods

for the measurement of catalytic
concentration of enzymes
Part 5.
IFCC
Method
for
Alkaline Phosphatase
(Orthophosphoric
–
Monoester Phospho
–
hydrolase, Alkaline Optimum, Ec 3.1.3.1).
J Clin Chem Clin Biochem
.

1983;21(11):

731
–
48.

18.

Tietz
NW
, Burtis
CA
, Duncan
P, Ervin K,
Petitclerc
CJ
, Rinker
AD
,
et al
. A
reference
method for measurement of alkaline
phosphatase activity in human serum.

Clin
Chem
.

1983;29(5):751
–
61.

19.

Gendler S. Uric
acid
. Kaplan A
,

et al
. Clin
Chem
the
Cv Mosby Co St Louis Toronto
Princeton
.

1984;1268
–
73.

Elsakka; IJBCRR,
12
(
2
):
1
–
9
, 20
16
; Article no.
IJB
C
RR
.
26029

9

20.

Mihara M, Uchiyama M. Determination
of
malonaldehyde precursor in tissues by
thiobarbituric acid test.
Anal Biochem
.

1978;86(1):271
–
8.

21.

Marklund
SL
. Superoxide
dismutase
isoenzymes in tissues and plasma from
new zealand black mice, nude mice
and
normal

Balb/C
mice
. Mutat Res
.

1985;148(1
–
2):129
–
34.

22.

Abdel
–
Bakky
MS
, Hammad
MA
, Walker
LA
, Ashfaq
MK
. Tissue
factor dependent
liver injury causes release of retinoid
receptors

(Rxr
–
Alpha And Rar
–
Alpha)
as
lipid droplets.

Biochem Biophys Res
Commun
. 201
1
;410(1):146
–
51.

23.

Ahsan R, Islam
KM
, Musaddik A, Haque E.
Hepatoprotective
activity of methanol
extract of some medicinal plants against
carbon tetrachloride induced hepatotoxicity
in albino rats
. Global J Pharmacol
.

2009;3(3):116
–
22.

24.

Kalu
FN
, Ogugua
VN
, Ujowundu
CO
,
Nwaoguikpe
RN
. Aqueous
extract of
combretum dolichopentalum leaf
–

A
potent
inhibitor of carbon tetrachloride induced
hepatotoxicity in rats
. Journal
of
Applied
Pharmaceutical Science
.

2011;1(10):114.

25.

Dalton
SR
, Lee
SM
, King
RN
, Nanji
AA
,
Kha
rbanda
KK
, Casey
CA
,
et al
. Carbon
Tetrachloride
–
induced liver damage in
asialoglycoprotein receptor
–
deficient mice.

Biochemical Pharmacology
.

2009;77(7):

1283
–
90.

26.

Manibusan
MK
, Odin M, Eastmond
DA
.
Postulated
carbon tetrachloride mode of
action:

A
review
.

J Environ Sci Health C
Environ Carcinog Ecotoxicol Rev
.

2007;25(3):185
–
209.

27.

Weber
LW
, Boll M, Stampfl A.
Hepatotoxicity
and mechanism of action of
haloalkanes
: Carbon
tetrachloride as a
toxicological model.

Crit Rev Toxicol
.

2003;33(2):105
–
36.

28.

Posen S, Doherty E. The
measurement of
serum alkaline phosphatase in clinical
medicine
. Adv Clin Chem
.

1981;22:163
–
245.

29.

Giannini
EG
, Testa R, Savarino V. Liver
enzyme alteration
: A
guide for clinicians
.
Canadian Medical Association Journal
.

2005;172(3):367
–
79.

30.

Kalu
FN
, Ogugua
VN
, Ujowundu
CO
,
Nwaoguikpe
RN
. Aqueous
extract of
combretum dolichopentalum leaf
–

A
potent
inhibitor of carbon tetrachloride induced
hepatotoxicity in rats
;

2011.

31.

Dalton
SR
, Lee
SM
, King
RN
, Nanji
AA
,
Kharbanda
KK
, Casey
CA
,
et al
. Carbon
tetrachloride
–
induced liver damage in
asialoglycoprotein receptor
–
deficient mice.

Biochem Pharmacol
.

2009;77(7):1283
–
90.

32.

Noori S, Rehman N, Qureshi M, Mahboob
T. Reduction
of carbon tetrachloride
–
induced rat liver injury by coffee and green
tea.

P
akistan Journal
of
Nutrition
.

2009;

8(4):452
–
8.

33.

Pierce
RA
, Glaug
MR
, Greco
RS
,
Mackenzie
JW
, Boyd
CD
, Deak
SB
.
Increased
procollagen mrna levels in
carbon tetrachloride
–
induced liver fibrosis
in rats.

J Biol Chem
.
1987;262(4):1652
–
8.

34.

Tamayo
RP
. Is
cirrhosi
s of the liver
experimentally produced by

Cc14
an
adequate model of human cirrhosis
?
Hepatology
.

1983;3(1):112
–
20.

35.

Herro R, Da Silva Ar, Aguilera Ar, Tamada
K, Croft M. Tumor
necrosis factor
superfamily

14 (Light)
controls thymic
stromal lymphopoietin to
drive pulmonary
fibrosis
. J Allergy Clin Immunol
.

2015;136(3):757
–
68.

36.

Herro R, Croft M. The
control of tissue
fibrosis by the inflammatory molecule light

(Tnf Superfamily Member 14). Pharmacol
Res
.

2015;104:151
–
5.

37.

Schuppan D. Structure
of the extracellular

matrix in normal and fibrotic liver
:
Collagens
and glycoproteins
. Semin Liver
Dis
.

1990;10(1):1
–
10.

______________________
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