Treatment with Markhamia tomentosa Benth. K. Schum prevents carbon tetrachloride-induced liver damage in rats

Abstract

Objective
Markhamia tomentosa (Bignoniaceae) is a medicinal plant with several pharmacological properties. However, its hepatoprotective effects have been little studied. The aim of this study was to evaluate the protective effects of the aqueous trunk bark extract of this plant against carbon tetrachloride (CCl 4 )-induced liver injury in rat.

Material and Methods
Thirty male albino Wistar rats were divided into six groups (five each) with Groups 1, 2, 3, and 4 as negative (distilled water), normal (distilled water), positive (silymarin 25 mg/kg), and plant extract (50 mg/kg) controls, respectively. Groups 5 and 6 were used as test groups and were given plant extract (25 or 50 mg/kg, respectively). Rats were pretreated once a day for 14 days orally with different substances. CCl 4 (0.5 mL/kg, i.p.) was administered on days 4 and 11 to all groups except Groups 1 and 4, to induce hepatitis. The rats were then sacrificed on day 15; liver functions and oxidative stress were assessed as well as histopathological changes.

Results
M. tomentosa extract significantly and dose dependently decreased alanine aminotransferase, aspartate aminotransferase, gamma-glutamine aminotransferase, total bilirubin, total cholesterol, and malondialdehyde values while increasing catalase, and glutathione values compared to the CCl 4 -treated group. Histological findings showed a reduction in necrosis and inflammatory cell infiltration in the liver while the lumen of distal and proximal tubes was improved in the kidney by the plant extract. These results may be due to some of the major bioactives compounds found in the aqueous extract.

Conclusion
These findings suggest that the aqueous extract of M. tomentosa may have liver protective effects through its antioxidant and anti-inflammatory mechanisms, supporting thereby its ethnomedicinal uses.

Full text

Adesh University Journal of Medical Sciences & Research • Article in Press | PB Adesh University Journal of Medical Sciences & Research • Article in Press | 1
Original Article
Treatment with Markhamia tomentosa Benth. K. Schum
prevents carbon tetrachloride-induced liver damage in rats
Romeo Joel Guemmogne Temdie1, Pierre Jidibe1, Borris Rosnay Tietcheu Galani1, Edwige Ymele Chiogo Vouo2,
Arnaud Doumogne Djasrane1, Emmanuel Le Fils Doumarsou Boumzina1, Selestin Dongmo Sokeng1, eophile Dimo2
1Department of Biological Sciences, University of Ngaoundere, Ngaoundere, Cameroon, 2Department of Animal Biology and Physiology, University of
Yaounde I, Yaounde, Cameroon.
*Corresponding author:
Romeo Joel Guemmogne Temdie,
Department of Biological
Sciences, University of
Ngaoundere, Ngaoundere,
Cameroon.
temdie2011@gmail.com
Received : 26July 2022
Accepted : 29November 2022
EPub Ahead of Print : 02 January 2023
Published :
DOI
10.25259/AUJMSR_54_2022
Quick Response Code:
INTRODUCTION
e liver is a major organ involved in the metabolism of endogenous or exogenous substances.[1,2]
is metabolic activity exposes the liver cells to the toxicity of xenobiotic and their metabolites
and thereby increases the risk of causing injury to liver, leading to liver inammation. Hepatitis,
an inammation of the liver tissue, can also be caused by infectious agents (bacterial, virus, and
parasites) or toxins that damage the liver cells.[3,4] During the process of liver injury, intracellular
substances leak out of injured cells, leading to the activation of neutrophils and monocytes, and
massive recruitment of these immune cells into the damaged liver tissue. Inammatory cells
are known to actively promote the production of reactive species, tumor necrosis factor-alpha
ABSTRACT
Objective: Markhamia tomentosa (Bignoniaceae) is a medicinal plant with several pharmacological properties.
However, its hepatoprotective eects have been little studied. e aim of this study was to evaluate the protective
eects of the aqueous trunk bark extract of this plant against carbon tetrachloride (CCl4)-induced liver injury in rat.
Material and Methods: irty male albino Wistar rats were divided into six groups (ve each) with
Groups1, 2, 3, and 4 as negative (distilled water), normal (distilled water), positive (silymarin 25 mg/kg), and
plant extract (50 mg/kg) controls, respectively. Groups5 and 6 were used as test groups and were given plant
extract (25 or 50mg/kg, respectively). Rats were pretreated once a day for 14days orally with dierent substances.
CCl4(0.5mL/kg, i.p.) was administered on days 4 and 11 to all groups except Groups1 and 4, to induce hepatitis.
e rats were then sacriced on day 15; liver functions and oxidative stress were assessed as well as histopathological
changes.
Results: M. tomentosa extract signicantly and dose dependently decreased alanine aminotransferase, aspartate
aminotransferase, gamma-glutamine aminotransferase, total bilirubin, total cholesterol, and malondialdehyde
values while increasing catalase, and glutathione values compared to the CCl4-treated group. Histological ndings
showed a reduction in necrosis and inammatory cell inltration in the liver while the lumen of distal and
proximal tubes was improved in the kidney by the plant extract. ese results may be due to some of the major
bioactives compounds found in the aqueous extract.
Conclusion: ese ndings suggest that the aqueous extract of M. tomentosa may have liver protective eects
through its antioxidant and anti-inammatory mechanisms, supporting thereby its ethnomedicinal uses.
Keywords: Liver diseases, Markhamia tomentosa, Hepatoprotective activity, Carbone tetrachloride, Antioxidant eect
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to remix, transform, and build upon the work non-commercially, as long as the author is credited and the new creations are licensed under the identical terms.
©2022 Published by Scientic Scholar on behalf of Adesh University Journal of Medical Sciences & Research
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Adesh University Journal of Medical
Sciences & Research Article in Press

Temdie, et al.: Markhamia tomentosa aqueous trunk bark extract prevents CCl4-induced liver injury
Adesh University Journal of Medical Sciences & Research • Article in Press | 2Adesh University Journal of Medical Sciences & Research • Article in Press | 3
(TNF-α), and other pro-inammatory cytokines that are
involved in the worsening of tissue lesions.[5]
Carbon tetrachloride (CCl4) is one of the chemical products
frequently used to induce liver injury.[6] e toxicity of
CCl4 results from its metabolism by cytochrome P450 in
the highly reactive species, trichloromethyl radical (CCl3),
and trichloromethyl peroxy radical (CCl3OO).[7] ese
molecules attack polyunsaturated fatty acids of biological
membranes and cause lipid peroxidation, which contributes
to severe cellular damages.[8,9] Destruction of mitochondria
membrane by CCl4 metabolites triggers caspase-3-
dependent apoptosis through specic cleavage of key cellular
proteins by caspase-3, a major cell death proteases.[10] It has
been proposed that the primary way to protect against CCl4-
induced liver damage is to inhibit the production of free
radicals.[11]
Medicinal plants have been used for centuries to treat diseases
due to their safety and ecacy, as well as their richness in
substances that could be used for therapeutic purposes or
as precursors to the synthesis of useful drugs.[12] In this
adventure, no part of the plant is le unexplored. Stem, roots,
leaves, owers, fruits, bark, and wood are all studied for their
medicinal potential.
Markhamia tomentosa is a medicinal plant of the
Bignoniaceae family found mainly in tropical regions.
[13] Ethnopharmacological data indicate its use against
headaches, canker, lumbago, chest pain, edema, rheumatic
pain, scrotal elephantiasis, anemia, diarrhea, backache, sore
eyes, intercostal pain, snakebite/venom, ailments of the
reproductive system, lung disorders, gout, bouts of swamp
fever, and external skin diseases.[14-17] A study revealed the
presence of alkaloids, tannins, saponins, anthraquinones,
cardiac glycosides, glycosides, phenols, and avonoids in
M. tomentosa methanol leaf extract and two compounds,
namely, 2-acetyl-6-methoxynaphtho[2,3-b] furan-4,9-dione
and 2-acetylnaphtho[2,3-b]furan-4,9-dione were identied
in the ethyl acetate leaf extract.[18,19] Based on these usages and
probably on phytochemical composition, the previous studies
demonstrated that M. tomentosa possessed antiprotozoal
and larvicidal activities, analgesic eects, acute and chronic
anti-inammatory eects, and antiarthritic eects.[18-22] It
was also proved that aqueous and methanol leaf extract
of M. tomentosa was relatively non-toxic to rats.[17] More
recently, it was found that the methanol leaf extract of M.
tomentosa exhibited a protective eect on D-galactosamine/
lipopolysaccharide-induced acute hepatitis.[23] However,
to the best of our knowledge, no research on the CCl4-
induced liver damage has been conducted. Furthermore,
the search for a viable natural remedy for liver diseases led
us to investigate the potential protective benets of aqueous
extract of M. tomentosa stem bark against CCl4-induced liver
injury in rats.
MATERIAL AND METHODS
Plant material
M. tomentosa bark was collected in July 2019, in the locality
of Bayangam, West region-Cameroon. e identication was
made by botanist of the National Herbarium of Cameroon,
in comparison with an existing voucher specimen registered
under the number 1974/SRFK. e harvested bark was cut
into small pieces that were dried in the shade. e dried bark
was crushed with an electric grinder and sieved, and the
powder was carefully stored in a plastic bottle for extraction.
Preparation of the aqueous extract
e plant extract was prepared as previously described by
Temdie et al.[19] Briey, 150 g of the M. tomentosa trunk
bark powder was introduced into the adiabatic device with
750 mL of hot distilled water (60°C) and the mixture was
stirred regularly for 24 h. e preparation was decanted
and ltered through Whatman No.3 paper. Two milliliters
of the ltrate were evaporated in an oven to determine the
extraction concentration (28mg/mL). e remaining ltrate
was stored at −20°C for later use.
Experimental animals
e study was conducted on male albino rats weighing
150 ± 10g. ese rats were reared under natural conditions
(room temperature, 12 h/12 h photoperiod, and average
humidity of 79 ± 10%) with free access to water and standard
food. e research was conducted in accordance with the
revised protocols for the care of laboratory animals (NIH
publication No.85-23, 1985) and the authorization granted
by the Cameroon National Ethical Committee (Reg. No.
FWAIRD 0001954).
Qualitative phytochemicals analysis
Qualitative phytochemical analysis of the aqueous extract of
M. tomentosa bark was performed to look for compounds
such as polyphenols, avonoids, tannins, anthraquinones,
saponins, anthocyanins, triterpenes, and alkaloids following
the protocol previously described by N’guessan et al.[24]
Experimental design
irty rats were divided into six groups of ve rats each made
up of four control groups (healthy, hepatitis, extract, and
positive control groups) and two test groups according to
treatment received [Table 1]. e animals were weighed on
the 1st day before treatment and then daily. Distilled water,
silymarin (Micro Labs Limited, Bengaluru, India), and plant
extract at the doses of 25 or 50 mg/kg were administered
orally (10mL/kg) at a daily dose for 14days. Liver injury was

Temdie, et al.: Markhamia tomentosa aqueous trunk bark extract prevents CCl4-induced liver injury
Adesh University Journal of Medical Sciences & Research • Article in Press | 2Adesh University Journal of Medical Sciences & Research • Article in Press | 3
induced aer 12h of fasting by intraperitoneal injection of
a mixture of an equal volume (1/1) of carbon tetrachloride
(Sigma-Aldrich, St. Louis, Missouri, USA) and olive oil
(Lesieur, 29, Quai Aulagnier 92665 Sur Seine Cedex,
France) at the dose of 0.5mL/kg, on days 4 and 11, except
for the healthy and extract control groups, which received
an equivalent volume of olive oil in the same manner.[25]
At the end of the treatment, the rats were sacriced under
anesthesia (ethylic ether) by decapitation. e blood sample
was collected in dry tubes and centrifuged at 3000rpm for
15 min to obtain serum. Liver and kidney were removed,
rinsed in normal saline solution, and weighed. e liver
and kidney sample was cold ground in Tris-HCl buer
(50 mM, pH7.4) to obtain a homogenate (20%), which was
centrifuged at 3000 rpm for 15 min. e supernatant was
recovered and stored as well as the serum at −20°C for the
biochemical analysis. Another sample of liver and kidney
was submerged in buered formalin (10%) for histological
analysis.[23]
Analysis of biochemical parameters
e determination of biochemical indicators such as
transaminases, gamma-glutamyl-transferase, triglycerides,
total protein, creatinine, total cholesterol, and total bilirubin
was performed according to the kit protocols (Chronolab
systems, Barcelona, Spain), revised in 2017. e analysis
of superoxide dismutase, malondialdehyde, catalase, and
reduced glutathione was done according to the Madoui
protocols.[26]
Histological analysis
Liver and kidney samples previously kept in buered
formalin (10%) were subjected to the following histological
techniques.[22] e samples were xed, dehydrated, and
impregnated. Aer impregnation, the samples were
then embedded in the paran to form blocks. A 50 µm
thick section of liver and kidney tissues was stained with
hematoxylin-eosin and observed under a light microscope
(ZEISS Axioskop, Paris, France).
Statistical analysis
Values are expressed as mean ± SEM. Statistical dierences
between groups were determined using analysis of variance
followed by Dunnett’s test. Data were analyzed using
GraphPad Prism (version 5.03). P ≤ 0.05 was considered
statistically signicant.
RESULTS
Qualitative phytochemical composition of the extract
e phytochemical test revealed the presence of polyphenols,
tannins, avonoids, saponins, anthocyanins, and
anthraquinones in the aqueous trunk bark extract [Tab l e 2].
Alkaloids and triterpenes were not found in the aqueous
trunk bark extract of M. tomentosa.
Body weight gain, liver and kidney absolute and relative
weights
e results show normal growth of healthy animals with a
weight gain in relation to the 1stday of treatment of 0.50 ±
3.43% on the 4thday and of 15.32 ± 2.10% on the 14thday
[Table 3]. Treatment of animals with CCl4 resulted in a
signicant loss of body mass (−1.11 ± 2.16 and −4.74 ±
2.91%), respectively, on the 4th and 14th days, compared to
the healthy control. Treatment with silymarin (25 mg /kg)
prevented body weight loss until the 4thday (0.45 ± 0.99%),
aer which weight loss was recorded until the 14th day
(−4.17 ± 1.51%). Pre-treatment with the extract (25mg/kg)
signicantly improved weight gain during treatment but
a signicant weight loss was registered at the end of the
experimental period (−2.49 ± 3.33%), compared to the
healthy control group. Administered alone, the extract did
not impair animal growth during the study as compared to
the healthy group.
Hepatitis control rats showed a slight reduction of the relative
liver mass (3.40 ± 0.13%) compared to healthy animals (3.50
± 0.06%). Silymarin treatment did not aect the relative liver
mass of rats compared to dierent controls groups (3.17
± 0.18%). A decrease in the relative mass of the liver was
recorded at the end of the study in animals treated with the
aqueous trunk bark extract of M. tomentosa. is reduction
was signicant at the dose of 50 mg/kg (3.02 ± 0.02%).
A signicant reduction was also observed aer treatment
of healthy animals with the plant extract (3.06 ± 0.07%), as
compared to control groups. At the end of the treatment,
no signicant variation in the relative kidney weight was
observed in dierent groups [Tab l e 3].
Tab l e 1: Distribution of the rats in the dierent groups in accordance with the treatment received.
Selected animals 30 rats
Groups formed Hepatitis control Healthy control Positive control Extract control Test groups
Treatment received H2O+CCl4H2O+Olive oil Sily 25+CCl4ATBE 50+Olive oil ATBE 50+CCl4ATBE 25+CCl4
Sily 25: Silymarin at a dose of 25 mg/kg. ATBE 50 or 25: Aqueous trunk bark extract of Markhamia tomentosa at the dose of 50 or 25 mg/kg.

Temdie, et al.: Markhamia tomentosa aqueous trunk bark extract prevents CCl4-induced liver injury
Adesh University Journal of Medical Sciences & Research • Article in Press | 4Adesh University Journal of Medical Sciences & Research • Article in Press | 5
Eect of the extract on enzymatic biochemical indicators
of liver function
e treatment of animals with CCl4 resulted aer 14days in a
signicant increase in the activity of alanine aminotransferase
(795.78 ± 15.78 U/L) compared to the healthy control
(189.30 ± 50.42 U/L). Treatment with silymarin (25mg/kg)
signicantly reduced ALT activity to 207.21 ± 31.68 U/L.
Plant trunk bark extract (25 or 50mg/kg) also reduced in a
signicant manner the ALT activity at 559.39 ± 91.69 U/L or
364.65 ± 39.00 U/L, respectively, compared to the hepatitis
control [Tab l e 4].
At the end of treatment, CCl4 caused a signicant increase
in aspartate aminotransferase activity (487.08 ± 57.92 U/L)
compared to the healthy control (273.11 ± 36.76 U/L).
Treatment with silymarin resulted in a signicant reduction
in AST activity (297.35 ± 20.89 U/L) compared to the
hepatitis control [Table 4]. e extract (25 or 50 mg/kg)
weakly decreased AST activity to 396.47±66.01 U/L or 353.79
± 41.57 U/L, respectively, compared to healthy control.
A signicant increase in the level of gamma-GT
(46.16 ± 4.32 U/L) was recorded on the 14thday, in animals
aer induction of hepatitis with CCl4 as compared to the
healthy control (17.59 ± 3.26 U/L). e treatment with
silymarin signicantly reduced the level of gamma-GT (23.60
± 2.87 U/L); likewise, the rats treated with the plant extract (25
or 50mg/kg) had a signicantly decreased gamma-GT activity
(21.74 ± 4.69 or 15.42 ± 1.48 U/L, respectively), compared
to the hepatitis control [Table 4]. M. tomentosa trunk bark
aqueous extract did not aect the activity of enzymatic
biochemical parameters of the liver function of healthy
animals (24.00 ± 1.88 U/L) as compared to healthy control.
Eect of the extract on non-enzymatic biochemical
indicators of liver and renal functions
Induction of hepatitis by CCl4 injection caused 2weeks aer
its administration, a signicant increase in the concentration
of total bilirubin (0.64 ± 0.09 mg/dL) compared to the
healthy control (0.31 ± 0.05mg/dL). Treatment of animals
with silymarin signicantly prevented the elevation of total
bilirubin concentration (0.27 ± 0.04mg/dL) compared to the
hepatitis control [Table5]. Administration of the extract (25
or 50mg/kg) signicantly reduced the level of total bilirubin
(0.29 ± 0.12 or 0.23 ± 0.00mg/dL, respectively) compared to
the hepatitis control.
No signicant increase in creatinine level was observed
between the healthy control (0.52 ± 0.01 mg/dL) and
hepatitis control (0.65 ± 0.06mg/dL). Silymarin (25mg/kg)
signicantly reduced creatinine levels (0.46 ± 0.01mg/kg)
compared to the hepatitis control. e extract (25 or
50mg/kg) did not signicantly inuence plasma creatinine
concentration (0.53 ± 0.04 or 0.56 ± 0.04 mg/dL,
respectively), compared to the hepatitis control [Tabl e 5]. In
the extract control group (50mg/kg), a signicant reduction
of creatinine level (0.47 ± 0.01 mg/kg) was observed,
compared to the hepatitis control.
e healthy control (5.33 ± 0.64mg/dL) and hepatitis control
(6.20 ± 0.51mg/dL) show no non-signicant variation in the
plasmatic level of total proteins, for in the case in rats treated
with the silymarin (25mg/kg), there is a signicant reduction
in the level of total proteins (3.40 ± 0.63mg/dL) compared
Tab l e 3: Changes in the body, liver and kidney weights of healthy and CCl4-treated rats under Markhamia tomentosa aqueous extract
treatment.
Treatment Body weight gain (%) Absolute weight (g) Relative mass (%)
Day 4th Day 8th Day 14th Liver Kidney Liver Kidney
H2O+CCl4−1.11±2.16#−1.40±2.30 −4.74±2.91#4.69±0.32 0.97±0.05 3.40±0.13 0.71±0.04
H2O+Olive oil 0.50±3.43* 0.33±4.09 15.32±2.10* 4.90±0.15 0.96±0.05 3.50±0.06 0.68±0.03
Sily 25+CCl40.45±0.99* −2.49±0.59 −4.17±1.51#4.05±0.20#0.88±0.05 3.17±0.18 0.68±0.03
ATBE 50+ Olive oil 3.25±0.12* 8.63±1.11#* 20.99±1.18* 5.02±0.15 1.01±0.02 3.06±0.07*## 0.61±0.01
ATBE 50+CCl44.68±0.74* 4.05±0.31* −2.49±3.33#4.14±0.05 0.95±0.02 3.02±0.02*## 0.69±0.01
ATBE 25+CCl45.11±2.43* 1.74±2.85 5.91±4.54#4.66±0.37 0.90±0.06 3.20±0.14 0.62±0.00
Each value represents the mean±SEM. n=5 number of animals in each group. *P<0.05 compared to hepatitis control group. #P<0.05 compared to the
healthy control group. Sily 25: Silymarin at a dose of 25 mg/kg. ATBE 50 or 25: Aqueous trunk bark extract of Markhamia tomentosa at the dose of
50 or 25 mg/kg, CCl4: Carbon tetrachloride.
Tab l e 2: Phytochemical composition of the aqueous extract of the
bark of Markhamia tomentosa.
Compounds found Observations
Polyphenols +
Tannins +
Flavonoids +
Anthraquinones +
Alkaloids −
Triterpenes −
Saponins +
Anthocyanins +
+: Present, −: Absent

Temdie, et al.: Markhamia tomentosa aqueous trunk bark extract prevents CCl4-induced liver injury
Adesh University Journal of Medical Sciences & Research • Article in Press | 4Adesh University Journal of Medical Sciences & Research • Article in Press | 5
to hepatitis control. e rats treated with the extract (25
or 50 mg/kg) as well as the extract control group show an
increase in the level of total proteins with a signicant eect
in the animals having received only the plant extract (9.27 ±
0.28mg/dL) compared to the healthy and hepatitis controls.
No signicant variation in triglyceride level was observed in
the dierent groups compared to the healthy and hepatitis
control.
e administration of CCl4 induced a signicant increase
in the total cholesterol level (80.72 ± 4.53 mmol/L) compared
to the healthy control (52.31 ± 6.38 mmol/L). Treatment with
the extract (25 or 50mg/kg) and silymarin (25mg) did not
signicantly reduce total cholesterol levels (79.72 ± 7.95;
75.37 ± 4.66; or 61.29 ± 5.93 mmol/L, respectively) compared
to the hepatitis control. Taken alone, M. tomentosa extract
induced a signicant increase in cholesterol levels (75.27 ±
0.84 mmol/L) compared to that of the healthy control.
Eect of the extract on hepatic and renal oxidative stress
CCl4 induced in animals a signicant increase in the level
of hepatic malondialdehyde (3.13 ± 0.01 nmol/mg of
protein) compared to the healthy control (2.48±0.01 nmol/
mg of protein). Treatment with silymarin (25 mg/kg) led
to a signicant reduction in the level of malondialdehyde
(2.36 ± 0.06 nmol/mg of protein). e treatment with an
aqueous extract (25 or 50mg/kg) resulted in a signicant
drop in the level of malondialdehyde (2.67 ± 0.03 or
2.79 ± 0.04 nmol/mg of protein, respectively) compared to
the hepatitis control [Table6]. e extract control animals
have a signicantly lower level of malondialdehyde (1.65
± 0.00 nmol/mg of protein) than the healthy control. CCl4
caused a signicant increase in the level of malondialdehyde
in the kidney (2.75 ± 0.02 nmol/mg of protein) compared to
the healthy control (1.23 ± 0.00 nmol/mg of protein). M.
tomentosa extract (25 or 50 mg/kg) signicantly reduced
malondialdehyde level to 1.65 ± 0.00 or 1.66 ± 0.05 nmol/
mg of protein, respectively, compared to the hepatitis
control.
e results show a signicant increase in catalase activity
(100.40±5.23 nmol/min/mg of protein) compared to the
healthy control (77.19 ± 5.10 nmol/min/mg of protein). e
extract at a dose of 25 or 50mg/kg induced a signicant rise in
catalase activity (136.08 ± 12.98 or 155.32 ± 12.82 nmol/min/mg
of protein) compared to the hepatitis control. e extract
induced a signicant elevation of hepatic catalase activity of
the extract control group (167.30 ± 6.26 nmol/min/mg of
protein), compared to the healthy and hepatitis control groups.
In the kidney, the extract (25 or 50mg/kg) caused a signicant
increase (141.85 ± 16.12 or 160.13 ± 9.51, nmol/min/mg of
protein, respectively) in catalase activity compared to the
hepatitis control (81.26 ± 9.53 nmol/min/mg of protein).
Treatment of healthy animals with the aqueous extract
of M. tomentosa result in signicant rise in renal catalase
activity (150.99 ± 11.73 nmol/min/mg of protein) compared
to control groups.
Tab l e 4: Fluctuations in enzymatic biochemical parameters of the
liver function of healthy and CCl4-treated rats under Markhamia
tomentosa aqueous extract treatment.
Treatments ALT (U/L) AST (U/L) 𝛾-GT (U/L)
H2O+CCl4795.78±15.78## 487.08±57.92## 46.16±4.32##
H2O+Olive
oil
189.30±50.42** 273.11±36.76** 17.59±3.26**
Sily 25+CCl4207.21±31.68** 297.35±20.89* 23.60±2.87**
ATB E
50+Olive oil
219.04±31.54** 301.70±18.60* 24.00±1.88**
ATBE
50+CCl4
364.65±39.00** 353.79±41.57 15.42±1.48**
ATBE
25+CCl4
559.39±91.69##* 396.47±66.01 21.74±4.69**
Each value represents the mean±SEM. n=5 number of animals in each
group. *P<0.05, **P<0.01, compared to the hepatitis control group,
##P<0.01 compared to the healthy control group. Sily 25: Silymarin at
a dose of 25 mg/kg. ATBE 50 or 25: Aqueous trunk bark extract of
Markhamia tomentosa at the dose of 50 or 25 mg/kg, CCl4: Carbon
tetrachloride.
Tab l e 5: Variations of non-enzymatic biochemical parameters of liver and renal functions of healthy and CCl4-treated rats under Markhamia
tomentosa aqueous extract treatment.
Treatments Total bilirubin
(mg/dL)
Creatinine
(mg/dL)
Total protein
(mg/dL)
Triglycerides
(mg/dL)
Total cholesterol
(mmol/L)
H2O+CCl40.64±0.09#0.65±0.06 6.20±0.51 166.26±9.65 80.72±4.53##
H2O+Olive oil 0.32±0.05* 0.52±001 5.33±0.64 130.09±11.60 52.31±6.38**
Sily 25+CCl40.27±0.04** 0.46±0.01** 3.40±0.63** 148.48±15.40 61.29±5.93
ATBE 50+Olive oil 0.35±0.05* 0.47±0.01* 9.27±0.28*## 151.67±32.34 75.27±0.84#
ATBE 50+CCl40.23±0.00** 0.56±0.04 6.68±0.12 116.86±14.13 75.37±4.66#
ATBE 25+CCl40.29±0.12* 0.53±0.04 6.65±0.72 149.87±19.73 79.72±7.95##
Each value represents the mean±SEM. n=5 number of animals in each group. *P<0.05, **P<0.01 compared to hepatitis control group; #P<0.05, ##P<0.01
compared to healthy control group. Sily 25: Silymarin at a dose of 25 mg/kg. ATBE 50 or 25: Aqueous trunk bark extract of Markhamia tomentosa at the
dose of 50 or 25 mg/kg, CCl4: Carbon tetrachloride.

Temdie, et al.: Markhamia tomentosa aqueous trunk bark extract prevents CCl4-induced liver injury
Adesh University Journal of Medical Sciences & Research • Article in Press | 6Adesh University Journal of Medical Sciences & Research • Article in Press | 7
Induction of hepatitis by CCl4 caused a signicant reduction
in the activity of superoxide dismutase in the liver tissue
(23.66 ± 2.91 U/mg of protein) compared to the healthy
control (49.16 ± 0.26 U/mg of protein). Silymarin (25mg/kg)
did not signicantly prevent (30.54 ± 5.77 U/mg of protein)
the reduction in superoxide dismutase activity compared to
the healthy control. Similarly, the extract (25 or 50 mg/kg)
did not rise signicantly (26.66 ± 2.51 or 28.52 ± 6.08 U/mg
of protein) the activity of superoxide dismutase compared to
the healthy control [Table6].
Hepatitis due to CCl4 caused a signicant reduction in
glutathione (267.64 ± 8.25 nmol/mg of protein) compared
to the healthy control (369.09±5.26 nmol/mg of protein).
Administration of silymarin (25 mg/kg) signicantly raised
glutathione levels in rats (346.81 ± 6.32 nmol/mg of protein).
e extract at a dose of 50 mg/kg caused a signicant
increase in the glutathione level (391.82 ± 30.61 nmol/mg of
protein) compared to the hepatitis control. An increase in the
glutathione level is noted in the extract control group (390.29
± 29.35 nmol/mg of protein) compared to the hepatitis
control [Tab l e 6].
Eect of extract on CCl4-induced liver inammation
Microphotography of the liver of a healthy rat shows normal
and uniformly stained hepatocytes with one or two nuclei.
Cells are well-organized in travel and the portal space is
clearly observable [Figure 1a]. e microphotography of
the liver of the CCl4-treated rat shows a remarkable invasion
of the portal space and the whole hepatic parenchyma
by mononuclear inammatory leukocyte cells. Edema
characterized by dilated capillaries could be found in
the parenchyma where hepatocytes present dark nuclei,
indicating chromatin condensation probably as a result of a
reduction in cellular activity due to inammation [Figure1b].
e liver section also shows cells necrosis represented by a
pile of cells nuclei without a plasma membrane surrounding
each. When the rats were treated with silymarin (25 mg/kg)
or aqueous extract (50 or 25 mg/kg), a clear improvement of
the histological parameters was observed compared to the
control and mononuclear inammatory leukocyte inltration
was also reduced [Figures1c-f].
Eect of extract on CCl4-induced kidney inammation
Microphotography of healthy control is normal [Figure 2a]. ere
is an important reduction of the lumen of the proximal and distal
convoluted tubes of CCl4-treated rats [Figure 2b] as compared to
the healthy animal. Renal section of rats treated with silymarin
(25mg/kg) or aqueous trunk bark extract of M. tomentosa (50 or
25mg/kg) was similar to that of healthy control [Figures2c-f].
DISCUSSION
is study was designed to evaluate the eects of aqueous trunk
bark extract of M. tomentosa on CC l4-induced liver injury in rats
since it has been claimed to be used for the management of liver
diseases. Results showed that aqueous extract of M. tomentosa
protects the liver against CCl4 damages. Administration of CCl4
occasioned a signicant increase in liver enzyme activities,
malondialdehyde levels, and induced an important leukocytes
inltration in liver tissue, which was prevented by treatment
with M. tomentosa aqueous trunk bark extract.
Inammation is a natural defense reaction of the body
against any aggression, whether exogenous or endogenous.
It takes place in the vascularized tissues and its ultimate
Tab l e 6: Changes in hepatic and renal oxidative stress indicators of healthy and CCl4-treated rats under Markhamia tomentosa aqueous
extract treatment.
Tissue Treatment MDA
(nmol/mg of protein)
Catalase
(nmol/min/mg of protein)
SOD
(U/mg of protein)
Glutathione
(nmol/mg of protein)
Liver H2O+CCl43.13±0.01## 100.40±5.23#23.66±2.91## 267.64±8.25##
H2O+Olive oil 2.48±0.01** 77.19±5.10* 49.16±0.26** 369.09±5.26**
Sily25+CCl42.36±0.06** 108.67±17.10#30.54±5.77#346.81±6.32*
ATBE50+Olive oil 1.65±0.00**## 167.30±6.26**## 33.37±7.79 390.29±29.35**
ATBE50+CCl42.79±0.04**## 155.32±12.82**## 28.52±6.08#391.82±30.61**
ATBE25+CCl42.67±0.03**## 136.08±12.98#26.66±2.51#241.47±9.17#
Kidney H2O+CCl42.75±0.02## 81.26±9.53 24.5±5.11#156.76±13.68
H2O+Olive oil 1.23±0.00** 67.32±7.71 42.25±6.07* 226.76±12.21
Sily25+CCl41.33±0.00** 74.05±18.87 38.29±2.83 190.58±1.44
ATBE50+Olive oil 1.63±0.03**## 150.99±11.73**## 36.37±2.52 207.94±31.60
ATBE50+CCl41.66±0.05**## 160.13±9.51**## 36.25±9.93 340.29±30.61**##
ATBE25+CCl41.65±0.00**## 141.85±16.12*## 37.08±1.14 310.88±19.97**#
Each value represents the mean±SEM. n=5 number of animals in each group. *P<0.05, **P<0.01 compared to hepatitis control group; #P<0.05, ##P<0.01
compared to healthy control group. Sily 25: Silymarin at a dose of 25 mg/kg. ATBE 50 or 25: Aqueous trunk bark extract of Markhamia tomentosa at the
dose of 50 or 25 mg/kg, CCl4: Carbon tetrachloride.

Temdie, et al.: Markhamia tomentosa aqueous trunk bark extract prevents CCl4-induced liver injury
Adesh University Journal of Medical Sciences & Research • Article in Press | 6Adesh University Journal of Medical Sciences & Research • Article in Press | 7
goal is protection against all dangers. e liver, being a
highly vascularized organ is, therefore, the privileged site of
inammatory reactions. It is a vital organ that is vulnerable to
factors of liver damage, such as toxics and their metabolites,
toxins, viruses, high-fat diet, and excessive alcohol
consumption.[27] Liver inammation caused by CCl4 is due to
the lyse of hepatic cells by free reactive radicals from CCl4
biotransformation. P450 cytochrome metabolized CCl4 into
CCl3, which, in turn, is processed in oxygenated condition
to the CCl3OO.[28] ese reactive species may cause harmful
eects in the body by oxidizing macromolecules such as
DNA, proteins, and polyunsaturated fatty acids. Hence,
prolonged exposure to such reactive CCl4 metabolites leads to
oxidative stress which results in the breakdown of membrane
structure, disruption of cell energy, and protein synthesis
processes, leading undoubtedly to cell injury.
Change in body weight is an adequate index to assess
the seriousness of pathologies and to judge the normal
functioning of the body. In addition, the ingestion of toxic
substances causes a loss of body weight.[29] However, the
interpretation of the results on the uctuation of the body
mass should not be done in an isolated way but it is necessary
to take into account the variation of the mass and the relative
mass of the organs. e results obtained show a signicant
decrease in the body mass of the hepatitis control animals
while the mass and the relative mass of their liver do not show
any signicant dierence compared to the healthy group,
thus demonstrating liver edema in the hepatitis control. e
rats treated with silymarin showed a signicant loss of weight
at the end of the treatment compared to healthy animals. e
average mass of their liver is likewise signicantly lower than
that of the healthy control, but not their relative mass. By
comparing these results with those of the hepatitis controls,
it appears that silymarin inhibited edema due to CCl4. e
results show that the hepatitis animals treated with the
extract of M. tomentosa show a markedly improved growth
compared to the hepatitis control rat, thus suggesting a
benecial eect of the extract against CCl4 hepatitis. At the
end of the study, the results indicate that the relative mass of
the liver of hepatitis animals treated with the extract is lower
Figure 1: Microphotography of the liver sections of healthy and
CCl4-treated rats under Markhamia tomentosa aqueous extract
treatment. Histological sections stained with hematoxylin-eosin
(×100). (a) Distilled water + CCl4, (b) distilled water + olive oil,
(c) silymarin (25mg/kg) + CCl4, (d) ATBE 50 + olive oil, (e) ATBE
50 + CCl4, and (f) ATBE 25 + CCl4. VP: Portal hepatic vein, He:
Hepatocytes, Cs: Sinusoidal capillary, Ah: Hepatic artery, Cb: Biliary
canaliculus, Il: Leukocyte inammation, NC: Necrosis cell, S:
Steatosis. ATBE 50 or 25: Aqueous trunk bark extract of Markhamia
tomentosa at the dose of 50 or 25mg/kg, CCl4: Carbon tetrachloride.
d
c
b
f
a
e
Figure2: Microphotography of the kidney sections of healthy and
CCl4-treated rats under Markhamia tomentosa aqueous extract
treatment. Histological sections stained with hematoxylin-eosin
(×100). (a) Distilled water + CCl4, (b) distilled water + olive oil,
(c) silymarin (25mg/kg) + CCl4, (d) ATBE 50 + olive oil, (e) ATBE 50
+ CCl4, and (f) ATBE 25 + CCl4. Eu: Urinary space, G: Glomerulus,
Tcd: Distal involved tube, Tcp: Proximal involved tube. ATBE 50 or
25: Aqueous trunk bark extract of Markhamia tomentosa at the dose
of 50 or 25mg/kg, CCl4: Carbon tetrachloride.
d
c
b
f
a
e

Temdie, et al.: Markhamia tomentosa aqueous trunk bark extract prevents CCl4-induced liver injury
Adesh University Journal of Medical Sciences & Research • Article in Press | 8Adesh University Journal of Medical Sciences & Research • Article in Press | 9
than that of healthy and hepatitis controls. is could suggest
an anti-edema eect of the plant extract. However, if we take
into consideration the growth of healthy rats treated with the
aqueous extract and their low relative liver weight, then we
could also suggest that the ability of plant components would
cause a reduction of the mass of hepatic tissue; and if so, the
mechanisms of this eect remain to be demonstrated.
Hepatitis caused by CCl4 is characterized by an increase in
plasma levels of ALT and AST.[30,31] Transaminases (ALT and
AST) are enzymes found in body cells, particularly in the
liver and muscles. An increase of these enzymes in the blood
is an indication of hepatic cells lysis.[30] Our results showed a
signicantly increased level of these enzymes when compared
to the healthy group. A substantial decrease level of these
enzymes was observed in hepatitis rats treated with aqueous
extract. ese results suggest that aqueous extract contains
compounds that may protect liver cells against CCl4 damage. In
general, polyphenol and avonoid compounds could prevent
cell lysis due to free radicals.[32,33] Gamma-glutamyltransferase
(𝛾GT) is a glycoprotein ubiquitous enzyme mostly found in the
renal, hepatic, and pancreatic cells. High levels of 𝛾GT in the
blood may be a sign of liver disease or damaged bile ducts.[34]
𝛾GT is more suitable for diagnosing cholangitis, cholecystitis,
and obstructive jaundice than alkaline phosphatase, leucine
aminopeptidase, and transaminases. Elevated serum value of
this enzyme is also observed in hepatitis induced by CCl4.[28]
Our results show signicant blood elevated activity of the 𝛾GT.
is nding is consistent with hepatobiliary disease induced by
CCl4. Serum 𝛾GT activity was signicantly reduced following
treatment of rats with the aqueous trunk bark extract of
M. tomentosa as well as silymarin thus proving the eciency of
the plant extract against CCl4-induced liver damage. Bilirubin
is a useful indicator to evaluate the excretory function and
dysfunction of hepatic bilirubin metabolism in the hepatic
cell.[35,36] Hepatocytes produce bilirubin from hemoglobin and
excrete it in bile or urine in a conjugate form with glucuronic
acid.[35,37] Bilirubin similarly leaks into the bloodstream
like transaminases when the biliary system is blocked.
Administration of CCl4 causes an inammatory reaction that
alters liver cells, disorganizes the hepatic parenchyma, and
increasing the plasma level of bilirubin.[35,37] Our results show
a signicant elevation of serum total bilirubin. Treatment
of animals with aqueous trunk bark extract of M. tomentosa
signicantly reduced blood bilirubin level. is result is in
agreement with that on 𝛾GT activity, suggesting that extract
has a protective eect against CCl4-induced hepatitis. Plant
avonoid compounds are a gied class of nutraceuticals
that have been reported to be able to protect against hepatic
damages.[30,38] ese results revealed an improvement of
enzymatic and non-enzymatic parameters thus demonstrating
the ecacy of the extract of trunk bark extract of M. tomentosa
to preserve normal functional liver status.
e endogen antioxidant defense system is composed of
superoxide dismutase, catalase, and glutathione peroxidase,
which are the main enzymes involved in the protection of the
body against oxidative stress, by keeping the redox balance
between the pro-oxidant and antioxidant mechanisms.[39] When
the antioxidant system failed to keep pro-oxidant processes
under control, this results in lipids peroxidation and alteration
of DNA and protein structures.[40] MDA is the end product of
lipid peroxidation widely used to quantify oxidative injury.[41]
Lipid peroxidation is one of the major features of the CCl4-
induced hepatitis.[41] A signicant elevation of MDA level in
the liver and kidney of hepatitis control rats was recorded in
the present study. ese results are indicative of the higher
level of lipid peroxidation in these tissues, signifying the failure
of endogen antioxidant defense mechanisms to prevent the
formation of excessive free radicals involved in cell damages.
is suggestion is consistent with the results which indicate a
signicant depletion in the tissues level of reduced glutathione
as a consequence of the reduction of the glutathione peroxidase
activity, and a signicant lowering of the SOD activity as
well. Glutathione peroxidase is a cytosolic enzyme that uses
reduced glutathione to catalyze the reduction of hydrogen
peroxide or peroxide radicals to water and oxygen or alcohols
and oxygen, respectively. is enzyme has a high anity to
H2O2 compared to catalase.[42] e peroxidase and reduced
glutathione are considered as major defense system when the
oxidative stress is not high.[43] is study revealed that treating
rats with an aqueous trunk bark extract of M. tomentosa
induce a signicant reduction in MDA levels, indicating that
this extract reduces membrane lipid peroxidation. e results
also demonstrate a substantial rise in catalase activity and
a decrease in glutathione rate. Hence, it is assumed that the
extract’s action may depend on the endogenous antioxidant
defense system. However, aer treating healthy animals with
an aqueous extract of M. tomentosa, an increased amount of
reduced glutathione was seen, as well as an increase in catalase
and SOD activity, resulting in a decreased level of MDA in
tissues. e ndings of this study demonstrated the preventive
ecacy of M. tomentosa aqueous extract against CCl4-induced
hepatic oxidative stress.
In this study, the histological alterations of liver tissue
recorded in rats subjected to CCl4 are consistent with the
ndings of earlier studies, which indicate a major lesion
of the hepatic parenchyma like hepatocytes necrosis and
inammatory leukocyte cells inltration in liver tissue as a
result of higher oxidative stress.
It has been shown that the inammatory reaction elicited
by CCl4 in liver tissue is caused by lipid peroxidation, which
increases the permeability of cell membranes and hence
induces cell death.[11] Furthermore, oxidative stress causes the
release of cathepsins B and then TNF-α by compromising the
integrity of the lysosome membrane.[11] All these processes,

Temdie, et al.: Markhamia tomentosa aqueous trunk bark extract prevents CCl4-induced liver injury
Adesh University Journal of Medical Sciences & Research • Article in Press | 8Adesh University Journal of Medical Sciences & Research • Article in Press | 9
according to Elgawish et al.,[11] result in an upregulation of
the pro-apoptotic protein p53, resulting in an inammatory
response in hepatic tissue. Early evidence suggested that
cytokines such as TNF-α, IL-1, IL-6, and IL-10 are involved
in the activation and enrolment of inammatory cells in the
liver.[27,44] e histological evaluation of the liver tissue revealed
a signicant decrease in the inltration of inammatory cells.
is nding is consistent with the antioxidant properties
early observed, like this, reveals the inhibiting eect of M.
tomentosa on CCl4-induced hepatitis. Based on the current
ndings, M. tomentosa aqueous extract may act similarly as
silymarin either by maintaining optimal redox stability in the
tissue through stimulation of antioxidant and non-enzymatic
molecules or by decreasing inammatory responses mainly
through mechanisms that need to be investigated.
e results of this study are consistent with those of Temdie
et al.[23] which demonstrated the protective activity of the
methanol leave extract of M. tomentosa on D-galactosamine/
lipopolysaccharide and those of Ibrahim et al.[45] which also
reported the prophylactic and therapeutic eects of a plant
of the family Bignoniaceae against paracetamol-induced liver
damage in rat. ese pharmacological activities are suggested
to be due to bioactive components of M. tomentosa trunk bark
aqueous extract such as polyphenols, avonoids, tannins,
saponins, and anthocyanins found in the aqueous trunk bark
extract. Some of these major classes of bioactive compounds
have been identied in the Markhamia genus[18,45] and their
strong antioxidant activities have been proved.[20,32,33]
CONCLUSION
e aqueous trunk bark extract of M. tomentosa has
hepatoprotective action against CCl4-induced liver
damage. Its antioxidant and anti-inammatory properties
would be responsible for this eect. Indeed, M. tomentosa
extract preserves the liver’s physiological functions and
architecture from the inammatory process caused by
CCl4 administration, by inhibiting lipid peroxidation,
improving the endogen antioxidant defense system, and
reducing inammatory cell mobilization in the injured
liver tissue. ese ndings support the use of M. tomentosa
by communities to treat liver diseases. Nonetheless, further
research is needed at this time to use this plant as an
alternative treatment for liver diseases.
Acknowledgments
e authors thank those who evaluated this research work at
all levels.
Authors’ contributions
All authors have participated to the realization of this
work either by conceiving and designing the experiments,
by conducting work on the eld and data collecting, by
contributing to data analysis, interpretation, and the
preparation of manuscript. e nal version of the article
was approved by authors for publication.
Declaration of patient consent
Patient’s consent not required as there are no patients in this
st udy.
Financial support and sponsorship
Nil.
Conicts of interest
ere are no conicts of interest.
REFERENCES
1. Opoku AR, Ndlovu IM, Terblanche SE, Hutchings AH.
In vivo hepatoprotective eects of Rhoicissus tridentata
subsp. cuneifolia, a traditional Zulu medicinal plant, against
CCl4-induced acute liver injury in rats. South Afr J Bot
2007;73:372-7.
2. Khan SA, Shahid S, Ahmad W, Ullah S. Pharmacological
importance of Clerodendrum genus: A current review. Int J
Pharm Sci Res 2017;2:22-30.
3. Lim CL, Suzuki K. Systemic inammation mediates the eects
of endotoxemia in the mechanisms of heat stroke. Biol Med
2016;9:1000376.
4. Jadon A, Bhadauria M, Shukla S. Protective eect of Terminalia
belerica Roxb. and gallic acid against carbon tetrachloride
induced damage in albino rats. J Ethnopharmacol
2007;109:214-8.
5. Taniguchi M, Takeuchi T, Nakatsuka R, Watanabe T,
Sato K. Molecular process in acute liver injury and regeneration
induced by carbon tetrachloride. Life Sci 2004;75:1539-49.
6. Weber LW, Boll M, Stamp A. Hepatotoxicity and mechanism
of action of haloalkanes: Carbon tetrachloride as a toxicological
model. Crit Rev Toxicol 2003;33:105-36.
7. Abdel-Kader MS, Abulhamd AT, Hamad AM, Alanazi AH,
Ali R, Alqasoumi SI. Evaluation of the hepatoprotective eect
of combination between hinokiavone and glycyrrhizin against
CCl4 induced toxicity in rats. Saudi Pharm J 2018;26:496-503.
8. Rahman MM, Muse AY, Khan DM, Ahmed IH, Subhan N,
Reza HM, et al. Apocynin prevented inammation and
oxidative stress in carbon tetra chloride induced hepatic
dysfunction in rats. Biomed Pharmacother 2017;92:421-8.
9. Domitrovie R, Skoda M, Marchesi VV, Cvijanović O, Pugel EP,
Stefan MB. Rosmarinic acid ameliorates acute liver damage
and brogenesis in carbon tetrachloride-intoxicated mice.
Food Chem Toxicol 2013;51:370-8.
10. Sun F, Hamagawa E, Tsutsui C, Ono Y, Ogiri Y, Kojo S.
Evaluation of oxidative stress during apoptosis and necrosis
caused by carbontetrachloride in rat liver. Biochim Biophys
Acta 2001;1535:186-91.
11. Elgawish RA, Rahman HG, Abdelrazek HM. Green tea extract

Temdie, et al.: Markhamia tomentosa aqueous trunk bark extract prevents CCl4-induced liver injury
Adesh University Journal of Medical Sciences & Research • Article in Press | 10 Adesh University Journal of Medical Sciences & Research • Article in Press | 11
attenuates CCl4-induced hepatic injury in male hamsters via
inhibition of lipid peroxidation and p53-mediatedapoptosis.
Toxicol Rep 2015;2:1149-56.
12. Adebayo AH, Abolaji AO, Kela R, Oluremi SO, Owolabi OO,
Ogungbe OA. Hepatoprotective activity of Chrysophyllum
albidum against carbon tetrachloride induced hepatic damage
in rats. Can J Pure Appl Sci 2011;3:1597-602.
13. Ghazanfar SA. Savanna Plants. An Illustrated Guide
(Illustrate). Basingstoke: Macmillan Educational Corps; 1989.
14. Bouquet A. Debray M. Plantes Medicinales de Cote d’Ivoire.
O.R.S.T.O.M. Paris: Oce de la Recherche Scientique
et Technique Outre-mer; 1974.
15. Arbonnier M. Arbres, arbustes et lianes des zones sèches
d’Afrique de l’Ouest. 2emeed. CIRAD, MNHN, Montpellier;
2000.
16. Burkill HM. e Useful Plants of West Tropical Africa. Kew:
Royal Botanic Gardens; 2002.
17. Temdie RJ, Kamdem GB, Minoue MG, Metchi FD, Kada AS,
Njiaza J, et al. Safety assessment of Markhamia tomentosa
(benth.) K. schum. (Bignoniaceae) leaves extracts, highlight
the psychostimulant eect of the methanol extract. JExp Appl
Trop Biol 2021;1:37-47.
18. Tantangmo F, Lenta BN, Boyom FF, Ngouela S, Kaiser M,
Tsamo E, et al. Antiprotozoal activities of some constituents of
Markhamia tomentosa (Bignoniaceae). Ann Trop Med Parasitol
2010;104:391-8.
19. Temdie GR, Fotio AL, Dimo T, Beppe J, Tsague M. Analgesic
and anti-inammatory eects of extracts from the leaves of
Markhamia tomentosa (benth.) K. schum. (Bignoniaceae).
Pharmacologia 2012a;3:565-73.
20. Adebajo AC, Aladesanmi AJ, Iwalewa EO, Akinkunmi EO,
Taiwo BJ, Olorunmola FO, et al. Antimicrobial and antioxidant
activities of some Nigerian medicinal plants. Afr J Tradit
Complement Altern Med 2006;4:173-84.
21. Temdie RJ, Fotio AL, Dimo T. Acute and chronic anti-
inammatory eects of the methanol leaf extract of Markhamia
tomentosa (benth.) K. schum. (Bignoniaceae). JSci Res Pharm
2012b;1:12-8.
22. Temdie GR, Metchi DM, Ymele E, Minoue K, Dimo T. e
eects of the methanol leaf extract of Markhamia Tomentosa
(benth.) K. schum. (Bignoniaceae) on arthritis induced by
complete freund ’s adjuvant in rats. World J Pharm Pharm Sci
2016;5:79-92.
23. Temdie RJ, Fotio AL, Metchi FD, Ymele EC, Tabi GT,
Dimo T. Protective activity of Markhamia tomentosa
(benth.) K. schum.(Bignoniaceae) methanol leaves extract
against D-galactosamine/lipopolysaccharide-induced acute
liver injury in mice. JBiosci Med 2020;8:74-89.
24. N’guessan JD, Zirihi GN, Kra AK, Kouakou K, Djaman AJ,
Guede-Guina F. Free radical scavenging activity, avonoid and
phenolic contents of selected Ivoirian plants. Int J Nat Appl Sci
2007;3:425-9.
25. Iredale J, Benyon R, Pickering J, McCullen M, Northrop M,
Pawley S, et al. Mechanisms of spontaneous resolution of
rat liver brosis. Hepatic stellate cell apoptosis and reduced
hepatic expression of metalloproteinase inhibitors. J Clin
Invest 1998;102:538-40.
26. Madoui S. Activités biologiques des extraits de Cytisus
Triorus. èse de doctorat en Biochimie, Département
de Sciences Biologiques, Université de Ferhat Abbas Sétif 1
d’Algérie; 2018.
27. Fotio AL, Nguepi MS, Tonfack LB, Temdie RJ, Nguelefack TB.
Acetaminophen induces liver injury and depletes glutathione
in mice brain: Prevention by Moringa oleifera extract. SAfr J
Bot 2020;129:317-23.
28. Ragab A, Nasr NE, Kahilo K. Protective eect of glycyrrhizc
acid against carbon tetrachloride-induced liver brosis in
rats: Role of Integrin subunit β like 1 (ITG β l1). Slov Vet Res
2019;56:673-9.
29. Boulila S, Elfeki A, Oudadesse H, Elfeki H. Substitution
eects of a carbonated hydroxyapatite biomaterial against
intoxication chloride nickel-exposed rats. Toxicol Mech
Methods 2015;25:155-65.
30. Kim TW, Lee DR, Choi BK, Kang HK, Jung JY, Lim SW, et al.
Hepatoprotective eects of polymethoxyavones against acute
and chronic carbon tetrachloride intoxication. Food Chem
Toxicol 2016;91:91-9.
31. Bruno R. Les dosages sanguins liés aux maladies hépatiques.
Centre Hépato-biliaire, Hôpital Universitaire Paul
Brousse-12-14 avenue Paul Vaillant Couturier-F-94800
Villejuif-Fance; 2016.
32. Gelen V, Şengül E, Gedikli S, Atila G, Uslu H, Makav M. e
protective eect of rutin and quercetin on 5-FU-induced
hepatotoxicity in rats. Asian Pac J Trop Biomed 2017;7:647-53.
33. Kumar BS, Sivaraj A, Elumalai EK, Kumar PV. Carbon
tetrachloride-induced hepatotoxicity in rats-protective role of
aqueous leaf extracts of Coccinia grandis. Int J Pharm Tech Res
2009;1:1612-5.
34. Payen JL, Robic MA. Conduite à tenir devant une élévation des
gamma-glutamyltransférases; 2007.
35. Sasidharan S, Aravindran S, Latha LY, Vijenthi R,
Saravanan D, Amutha S. In vitro antioxidant activity
and hepatoprotective eects of Lentinula edodes
against paracetamol-induced hepatotoxicity. Molecules
2010;15:4478-89.
36. Al-Harbi NO, Imam F, Nadeem A, Al-Harbi MM, Iqbal M,
Ahmad SF. Carbon tetrachloride-induced hepatotoxicity in rat
is reversed by treatment with riboavin. Int Immunopharmacol
2014;21:383-8.
37. Ajith A, Hema U, Aswathy MS. Zingiber ocinale roscoe
prevents acetaminophen-induced acute hepatotoxicity by
enhancing hepatic antioxidant status. Food Chem Toxicol
2011;45:2267-72.
38. Tiwary BK, Dutta S, Dey P, Hossain M, Kumar A, Bihani S, et al.
Radical scavenging activities of Lagerstroemia speciosa (L.) pers.
Petal extracts and its hepato-protection in CCl4-intoxicated
mice. BMC Complement Altern Med 2017;17:55.
39. Pieme CA, Tatangmo JA, Simo G, Nya PC Moor VJ,
Moukette BM, et al. Relationship between hyperglycemia,
antioxidant capacity and some enzymatic and non-enzymatic
antioxidants in African patients with Type2 diabetes. BMC
Res Notes 2017;10:141.
40. Su JJ, Wang X, Song W, Bai X, Li C. Reducing oxidative stress
and hepatoprotective eect of water extracts from Pu-erh
tea on rats fed with high-fat diet. Food Sci Human Wellness
2016;5:199-206.

Temdie, et al.: Markhamia tomentosa aqueous trunk bark extract prevents CCl4-induced liver injury
Adesh University Journal of Medical Sciences & Research • Article in Press | 10 Adesh University Journal of Medical Sciences & Research • Article in Press | 11
41. Basu S. Carbon tetrachloride-induced lipid peroxidation:
Eicosanoid formation and their regulation by antioxidant
nutrients. Toxicolology 2003;189:113-27.
42. Higuchi M. Antioxidant properties of wheat bran against
oxidative stress. Wheat and rice in disease prevention and
health benets, risks and mechanisms of whole grains in health
promotion; 2014.
43. Tabet F, Touyz MR. Reactive oxygen species, oxidative stress,
and vascular biology in hypertension. Comprehensive
Hypertension; 2007.
44. Yan M, Huo Y, Yin S, Hu H. Mechanisms of acetaminophen-
induced liver injury and its implications for therapeutic
interventions. Redox Biol 2018;17:274-83.
45. Ibrahim MB, Kaushik N, Sowemimo AA, Odukoya OA.
Review of the phytochemical and pharmacological studies of
the genus Markhamia. Pharmacogn Rev 2016;10:50-9.
How to cite this article: Temdie RJ, Jidibe P, Galani BR, Vouo EY,
Djasrane AD, Boumzina EL, et al. Treatment with Markhamia tomentosa
Benth. K. Schum prevents carbon tetrachloride-induced liver damage in
rats. Adesh Univ J Med Sci Res, doi: 10.25259/AUJMSR_54_2022