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Research Article
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Evaluation of the Hepatoprotective Effect of Fumaria parviflora and Momordica balsamina from Saudi Folk Medicine Against Experimentally
Induced Liver Injury in Rats |
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Saleh I. Alqasoumi,
Mohammad S. Al-Dosari,
Abdulmalik M. AlSheikh
and
Maged S. Abdel-Kader
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ABSTRACT
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In a project to evaluate the efficacy of traditional
Saudi plants used for liver problems the two plants Fumaria parviflora
Lam. (Fumariaceae) and Momordica balsamina Linn. (Cucurbitaceae)
were studied. The ethanol extract of the aerial part of Fumaria parviflora
and the leaves of Momordica balsamina were subjected to hepatoprotective
assays using Wistar albino rats. Liver injury induced in rats using carbon
tetrachloride. The biochemical parameters; serum glutamate oxaloacetate
transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), alkaline
phosphatase (ALP) and total bilirubin were estimated as reflection of
the liver condition. Based on the results of the biochemical parameters
measurements, histopathological study was performed on the liver of rats
treated with two extracts. The normal appearance of hepatocytes indicated
a good protection of the extracts from carbon tetrachloride hepatotoxicity.
All the results were compared with silymarin, the reference hepatoprotective
drug.
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How
to cite this article:
Saleh I. Alqasoumi, Mohammad S. Al-Dosari, Abdulmalik M. AlSheikh and Maged S. Abdel-Kader, 2009. Evaluation of the Hepatoprotective Effect of Fumaria parviflora and Momordica balsamina from Saudi Folk Medicine Against Experimentally
Induced Liver Injury in Rats. Research Journal of Medicinal Plants, 3: 9-15. DOI: 10.17311/rjmp.2009.9.15 URL: https://scialert.net/abstract/?doi=rjmp.2009.9.15
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INTRODUCTION
Natural products still represents a vital source for biologically active drugs
with unique mechanisms of actions. Many traditional plants are used for the
treatment of liver problems. Study of these led to the discovery of active compounds
yet developed to successful drugs. Silymarin (Saller et
al., 2007), schisandrin B (Zhu et al., 1999;
Cyong et al., 2000), phyllanthin, hypophyllanthin,
picroside I and kutkoside (Ram, 2001) are examples of
natural antihepatotoxic compounds derived from traditional herbs. In Saudi Arabia,
many plants are claimed to treat liver problems (Mossa et
al., 1987), however, most of these claims lack the scientific prove.
Here was an interest in screening of such plants for their hepatoprotective
effect and in advanced steps identify their active constituents. The aerial
parts of Fumaria parviflora are used in Saudi folk medicine for bile
secretion, spleen disorders and jaundice. Other used includes diaphoretic, diuretic,
anthelmintic and laxative. It is also claimed to have antimicrobial activity
so used for treatment of bacterial and skin diseases (Mossa
et al., 1987; Ghazanfar, 1994; El-Shanawany,
1996). Extract of the aerial parts of Fumaria parviflora showed significant
anticholinesterase activity (Orhan et al., 2004).
It also proved to be active against larvae of Haemonchus contortus (Hordegen
et al., 2006). Pharmacological studies indicated that Fumaria parvi
posses anti-inflammatory, anti-nociceptive and antidiabetic activities (Akhtar
et al., 1984; Heidari et al., 2004). Different
parts of Momordica balsamina are used as galactagogue, anthelmintic,
laxative and tonic. It is also used for disorders of the liver (Mossa
et al., 1987; El-Shanawany, 1996). Recent study
indicated that the fruit pulp extract of Momordica balsamina has promising
anti-HIV, antimicrobial and antiplasmodial effects (Bot et
al., 2007; Benoit-Vical et al., 2006;
Jigam et al., 2004).
MATERIALS AND METHODS
Plant Materials
The aerial parts of Fumaria parviflora Lam. (Asteraceae)
and the leaves of Momordica balsamina Linn. (Cucurbitaceae)
were purchased from the local market in Riyadh, in April 2003. The plants
were identified by Dr. Mohammad Atiqur Rahman, taxonomist of the Medicinal,
Aromatic and Poisonous Plants Research Center (MAPPRC), College of Pharmacy,
King Saud University, Riyadh, Saudi Arabia.
Extraction
The dried ground aerial parts of Fumaria parviflora and leaves
of Momordica balsamina (50 g from each plant) were extracted to
exhaustion by percolation at room temperature with 90% ethanol and the
extract was evaporated in vacuo to leave 7.4 and 6.3 g of residue.
Animals
Wistar albino rats (150-200 g) of either sex roughly the same age (8-10
weeks), obtained from the Experimental Animal Care Center, College of Pharmacy,
King Saud University, Riyadh were used. The animals were housed under constant
temperature (22±2 °C), humidity (55%) and light/dark conditions (12/12
h). They were provided with Purina chow and free access to drinking water
ad libitum (Abdel-Kader and Alqasoumi, 2008). The
experiments and procedures used in this study were approved by the Ethical Committee
of the College of Pharmacy, King Saud University.
Chemicals
Silymarin (Sigma Chemical Company, USA).
Hepatoprotective Activity
Male Wistar rats were divided into five groups six animals each. Group
1 was kept as a control group. Groups 2, 3, 4 and 5 received 0.125 mL
of CCl4 in liquid paraffin (1:1) per 100 g body weight intraperitoneally.
Group 2 received only CCl4 treatment. Group 3 was administered
silymarin at a dose of 10 mg kg-1 p.o. Groups 4 and 5 were
treated with 250 and 500 mg kg-1 of extracts respectively.
Drug treatment was started 5 days prior to CCl4 administration
and continued till the end of the experiment. After 48 h, following CC14
administration the animals were sacrificed using ether anesthesia. Blood
samples were collected by heart puncture and the serum was separated for
evaluating the biochemical parameters. The liver was immediately removed
and a small piece was fixed in 10% formalin for histopathological assessment.
Determination of the Enzyme Levels
The biochemical parameters such as serum glutamate oxaloacetate transaminase
(SGOT), serum glutamate pyruvate transaminase (SGPT), alkaline phosphatase (ALP)
and total bilirubin were estimated by reported methods (Boon
et al., 2006). The enzyme activities were measured using diagnostic
strips (Reflotron®, ROCHE) and were read on a Reflotron®
Plus instrument (ROCHE).
Statistical Analysis
For each set of experiments where two or more than two groups were compared,
an analysis of variance (ANOVA) test was used to determine the significance
of the differences. Differences between the control and CCl4-treated
group were compared for significance using student`s t-test for non paired samples
(Woolson and Clarke, 2002). All the values shown are the
mean±SE mean.
Histopathology
The livers of treated animals were immediately removed and a small piece
was fixed in 10% formalin for histopathological assessment. All specimens were
placed in cassettes and loaded into tissue baskets. The specimens were subjected
to dehydration, clearing and inflatration by immersion in different conc of
ethanol (70-100%), xylene (3 times, 1 h each) and finally paraffin wax (4 times,
1 h each). The tissues were then transferred into moulds filled with paraffin
wax. After orienting the tissues by hot forceps the moulds were chilled on cold
plates and excess wax were trimmed off using a knife. The rotary microtome (Leitz
1512) was used for making thin sections (3°m). The sections were placed onto
clean slides that were drained vertically for several minutes before placing
them onto a warming table at 37-40°C (Prophet et al.,
1994). The slides were then deparaffinized, hydrated and to stained in Mayer`s
hematoxylin solution for 15 min. The slides were then washed in lukewarm running
tap water for 15 min, placed in distilled water, 80% ethyl alcohol for 1 to
2 min then counterstained in eosin-phloxine solution for 2 min. The slides were
then dehydrated and cleared through 2 changes each of 95% ethyl alcohol, absolute
ethyl alcohol and xylene, 2 min each and finally mounting with resinous medium.
RESULTS AND DISCUSSION
Male Wistar rats were used to estimate the serum glutamate oxaloacetate transaminase
(SGOT), serum glutamate pyruvate transaminase (SGPT), alkaline phosphatase (ALP)
and total bilirubin serum enzymes as indicator for liver protection against
carbon tetrachloride hepatotoxicity. Treatment of animals with the hepatotoxic
agent carbon tetrarchloride resulted in significant increase of transaminases
(SGOT and SGPT) and alkaline phosphate levels (ALP) due to hepatocytes damage
(Zafar and Ali, 1998). Sever jaundice was reflected by
increase level of serum bilirubin (Lin et al., 1997)
(Table 1).
Treatment of animals with the known hepatoprotective agents silymarin as a
reference standard resulted in significant decrease in the elevated levels of
SGOT, SGPT, ALP and bilirubin (p<0.001) (Table 1). Silymarin
act as an antioxidant by scavenging prooxidant free radicals and by increasing
the intracellular concentration of GSH. It also exhibits a regulatory action
of cellular membrane permeability and increase in its stability against xenobiotics
injury; increasing the synthesis of ribosomal RNA by stimulating DNA polymerase-I,
exerting a steroid like regulatory action on DNA transcription and stimulation
of protein synthesis and regeneration of liver cells (Saller
et al., 2007; Dehmlow et al., 1996a,
b; Gakova et al., 1992).
Silymarin efficacy is not limited to the treatment of toxic and metabolic liver
damage; it is also effective in acute, chronic hepatitis and in inhibiting fibrotic
activity (Hikino and Kiso, 1988; Mayer
et al., 2005).
Treatment of the animals with doses of 500 mg kg-1 body weight
of Fumaria parviflora aerial parts ethanol extract prior to CCl4
administration resulted in decrease in the levels of SGOT (24.1%), SGPT
(21.6%), ALP (20.5%) and bilirubin (11.3%) levels. The lower dose of the
extract (250 mg kg-1, body weight) show only significant decrease
of the levels of APL (15.2%) (Table 1).
Table 1: |
Effects of ethanolic extracts of Fumaria parviflora
and Momordica balsamina on serum biochemical parameters |
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*p<0.05; **p<0.01; ***p<0.001, aas
compared with the normal saline (control) group; bas compared
with the CCl4 only group |
Momordica balsamina (Leaves) crude extract when given to rats
in the two used doses (250 and 500 mg kg-1, body weight) before
CCl4 treatment resulted in highly significant (p<0.001)
decrease in the levels of SGOT, SGPT, ALP and bilirubin (Table
1). The obtained results indicated a high degree of protection against
the hepatotoxic effect of CCl4.
The histological appearance of the hepatocyte reflects their conditions
(Fig. 1A). Exposure of hepatocytes to toxic agents such
as CCl4 leads to histopathological changes from the normal
cell appearance. The hepatocytes of rat livers treated with a single dose
of 1.25 mL CCl4 kg-1, showed centrilobular hepatocyte
necrosis, microvesicular fatty change and extensive fatty change were
observed on the midzonal or entire lobe at 24 h (Fig. 1B).
Liver tissue of rats treated with CCl4 and silymarin showed
good recovery with absence of necrosis, fatty depositions and recovery
to normal histological appearance of hepatocytes (Fig. 1C).
The central vein has minimal portal inflammation. Histological appearance
of rat liver treated with 500 mg kg-1 Fumaria parviflora
aerial parts total extract showed minimal portal inflammation and more
or less normal appearance (Fig. 1D). Sections of the
rat liver treated with CCl4 and Momordica balsamina
(500 mg kg-1) showed significant recovery with disappearance
of fatty deposition and necrosis. The portal tracts appeared almost clear
indicating a potent hepatoprotective activity (Fig. 1E).
The current study aimed to evaluate the effect of Fumaria parviflora
and Momordica balsamina as protective for hepatocytes. The higher
dose of Fumaria parviflora showed significant decrease in the level
of biochemical markers as well as notable recovery of the histological
characters of the hepatocytes. Momordica balsamina showed superior
protective effect in both biochemical markers assessment and histopathological
appearance of hepatocytes.
Fig. 1: |
Histopathological appearance of liver cells, (A) normal
cells, (B) liver cells of rats treated with CCl4, (C) liver
cells of rats treated with CCl4 and silymarin, (D) liver
cells of rats treated with CCl4 and Fumaria parviflora,
(E) liver cells of rats treated with CCl4 and Momordica
balsamina |
ACKNOWLEDGMENTS
The authors are grateful to Mr. Ali D. Al-Sawadi and Mr. Malik Sawood
for technical assistance. The research was supported by grants from King
Abdul Aziz City of Science and Technology (KACST), Riyadh, Saudi Arabia
to Dr. Alqasoumi.
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