Antioxidant Activity and Biological Evaluation of Hot-water Extract of Artemisia monosperma and Capparis spinosa Against Lead Contamination
Hot-water extracts of Capparis spinosa and Artemisia monosperma were studied against lipid peroxidation induced by lead acetate in rats. Ten rats were assigned to each of four groups. Group 1 was negative control, group 2 received only lead acetate (0.6%) in drinking water (positive control), group 3 received hot-water extract from Artemisia (5%) with lead acetate in drinking water (0.6%) and group 4 received hot-water extract from Capparis (5%) with lead acetate in drinking water (0.6%) experiment continued for 6 weeks before scarifying the animals and blood samples were collected. Chemical composition and antioxidant activities of hot-water extract from Artemisia and Capparis sp. were estimated using chemical methods. Total Antioxidants in Artemisia and Capparis extracts were 675.33, 115.66 μmol Trolox per 100 mL, respectively. Activity of the antioxidant enzyme glutathione-S-transferase activity decreased after lead administration but extracts from Artemisia and Capparis maintained the level to normal. Lead administration resulted in significant increases in the concentration of serum triglycerides, urea and aspartate transaminase, alanine transaminase (AST and ALT). Extracts from Artemisia and Capparis reduced the elevated concentrations to normal values. The obtained results showed that the biochemical alterations produced by lipid peroxidation after lead administration returned to normal values or even improved due to high levels of total antioxidants in extracts of Artemisia and Capparis sp. Data concluded that the protective effects of hot-water extracts from Artemisia and Capparis sp. may play a role in protection against lead acetate in rats. This protective effect may be due to high level of total antioxidant contents in these plants.
to cite this article:
A. Al-Soqeer , 2011. Antioxidant Activity and Biological Evaluation of Hot-water Extract of Artemisia monosperma and Capparis spinosa Against Lead Contamination. Research Journal of Botany, 6: 11-20.
Received: June 15, 2010;
Accepted: October 17, 2010;
Published: April 09, 2011
In recent years the therapeutic importance of herbal drugs has gained considerable attention. It has been found that utilization of cheap plant sources that are without side effects is a trend in all countries.
In Saudi Arabia herbal medicine is widely spread and many local plants are
used for treatment of various ailments. One of these plants is Capparis spinosa
(CS) (Capparidaceae), locally known as Shafalah. Scientific classification and
uses of Capparis spinosa are shown in Fig. 1. They
are native shrubs widely distributed in Al-Qassim region. This plant was found
to be common to the Mediterranean basin and has been used by the traditional
medicine for its diuretic and antihypertensive effects and also in certain pathological
conditions related to uncontrolled lipid peroxidation. The extract contains
many constituents, in particular some flavonoids (kaempferol and quercetin derivatives)
and hydrocinammic acids with several known biological effects such as the anti-inflammatory
and the antioxidant ones (Panico et al., 2005).
|| Scientific classification and uses of Capparis spinosa
The phytotherapy knowledge of this plant has been of interest to many investigators.
The main constituents of CS have been demonstrated by other workers to be flavonoids,
alkaloids, lipids and glucosinolates and these constituents may vary from one
locality to another (Calis et al., 2002; Sharaf
et al., 2000; Brevard et al., 1992).
Shafalah has been used since ancient times in Morocco in the treatment of digestive
disorders and was commonly used in phytomedicine as an antibacterial (Singh
et al., 2002), antiulcerogenic (Khayyal et
al., 2001) antiproliferative (Nakano et al.,
Recently, Yang et al. (2010) found that fruits
of Capparis spinosa contain a significant amount of compounds with many
health benefits. Three new alkaloids, (1) Capparisine A, (2) Capparisine B,
(3) Capparisine C and (4) two known alkaloids, (4) 2-(5-hydroxymethyl-2-formylpyrrol-1-yl)
propionic acid lactone and (5) N-(3'-maleimidy1)-5-hydroxymethyl-2-pyrrole formaldehyde
were isolated from the fruits of C. spinosa.
Another important plant is Artemisia species. Scientific classification and uses of Artemisia monosperma (Athir) are presented in Fig. 2.
The herb, Artemisia monosperma, is a perennial fragrant plant which
grows widely and wildly in the Arabian deserts. The leaves of the plant are
taken in folk medicine by certain women of Jordan for abortion induction (Hijazi
and Salhab, 2010). In a phytochemical study of Artemisia monosperma
three compounds were characterised by spectroscopic means. Compounds were evaluated
for their ability to inhibit 12-lipoxygenase, as antibacterials and in a cytotoxicity
assay (Stavri et al., 2005).
The antioxidant activities of various fractions of Artemisia species
were investigated, especially, the n-butanol extract that was found to causes
significant increase in the rat liver cytosolic superoxide dismutase (SOD) and
catalase activities (Juteau et al., 2002; Kim
et al., 2003). The Artemisia oil has a characteristic flavor,
due to the presence of many components with strong sensory properties at a low
threshold, such as trans-ethyl cinnamate (20.8%) and thus could be suitable
for using as antioxidant and flavoring agent in the food industry (El-Massry
et al., 2002).
|| Scientific classification and uses of Artemisia monosperma
It was claimed by many researchers that the risk from contamination by heavy
metals is mainly due to the oxidative stress produced by these metals and consequently
depletion of body antioxidants (Ercal et al., 2001).
Because many synthetic antioxidants (butylated hydroxyanisole and butylated
hydroxytoluene) have been proved to have undesirable side effects (Jayaprakasha
et al., 2007). There has been an increasing interest in the use of
medicinal plants rich in antioxidants to reduce free radicals-induced tissue
injury (Zhishen et al., 1999). Oxidative stress
is the excess formation of and/or incomplete removal of highly reactive molecules
such as Reactive Oxygen Species (ROS). In vivo some of ROS play positive
role such as energy production, phagocytosis and regulation of cell growth (Halliwell
and Gutteridge, 1999). On the other hand, ROS are capable of damaging a
wide range of essential biomolecules such as proteins, DNA and lipids (Farber,
1994). Cells have several antioxidant defense mechanisms that help to prevent
the destructive effects of ROS. These defense mechanisms include anti oxidative
enzymes, such as superoxide dismutase, catalase and glutathione peroxidase and
of small molecules such as glutathione and vitamin C and E (Maser
et al., 2002). The efficiency of the antioxidant defense system is
altered under pathological conditions and therefore the ineffective scavenging
and/or over production of free radicals may play a crucial role in determining
tissue damage (Aruoma, 1994; Halliwell,
From the foregoing considerations, it can be concluded that Phyto chemicals content in plants are promising alternatives to synthetic drugs provided that certain precautions were taken in consideration. The present investigation aims at analyzing the chemical composition and nutritive value of Capparis spinosa and Artemisia monosperma and test their efficacy against lead contamination in rats.
MATERIALS AND METHODS
The study to be described was carried out at the College of Agriculture and Veterinary Medicine, Qassim university, Saudi Arabia during the period 2009/2010.
Capparis spinosa (CS): Scientific classification and uses
of Capparis spinosa is shown in Fig. 1. Scientific
classification and uses of Artemisia monosperma (Athir) is shown in Fig.
Preparation of pressurized hot water extraction: Hot water extraction
of Artemisia and Capparis were prepared according the methods
described by Abdel-Salam et al. (2009): Approximately
50 g of each of Artemisia and Capparis materials were cut into
small pieces, melded and placed in a flask (2 L) with 1000 mL of distilled water
and boiled for 15 min, then the mixture was filtered twice, first through cheese-cloth
(50% cotton/50% polyester) and then through filter paper (Whatman No. 2). The
final concentrations of the prepared of Artemisia or Capparis
were 5% as a total solids. The amounts of obtained Artemisia and Capparis
extracts were preserved in sterile dark bottles (500 mL) in a cool environment
(4°C) until further use.
Antioxidant activity determination in Artemisia monosperma and Capparis
spinosa: Antioxidant activity was determined according to the method
of Sanchez-Moreno et al. (1998). Antioxidant capacity
was quantified as μg Trolox equivalents using a standard curve.
Proximate analysis and nutritive value: The vitamin C content was determined
by the titrimetric method according to the method of AOAC
(1990). Lipid content of these plants was determined by the direct extraction
method. Three grams of dried sample was extracted by 50 mL of petroleum ether
at 60°C for 60 min in a soxtec system HT (series 1043 Extractor unit, Tecator).
The lipid value was obtained by mean of weighing. Protein content was determined
by the Kjeldahl method. Other determinations were carried out according to Benjapak
et al. (2008).
Animals: Forty male Swiss albino rats weighting about 120-150 g were randomly divided into four-test groups (each containing 10 rats). Animals were placed in cages and were assigned to specific diets.
Diets: The composition of the basal diet used in this study is as follows:
milk protein (12%), sucrose (5%), fat (10%), vitamin mixtures (1%), salt mixtures
(4%), fiber (4%) and starch (64%). Rats were divided into the following groups.
||The first group (G1) was fed on the basal diet.
without lead acetate (control negative)
||The second group (G 2) was fed on the basal diet with lead
acetate in drinking water (0.6%) (control positive)
||The third group (G3) was fed on basal diet + hot-water extract
from Artemisia (5%) with lead acetate in drinking water (0.6%)
||The fourth fed (G 4) on basal diet + hot-water extract from
capparis (5%) with lead acetate in drinking water (0.6%)
The feeding experiment continued for 6 weeks and at the end of this period,
rats were anesthetized by diethyl ether, bled and sacrificed. Blood samples
were collected and frozen at (4-6°C) for 2 h before centrifugation at 3000X
g for 30 min. Sera were harvested, labeled and stored deep frozen (-20°C)
until used for assays.
Biochemical analysis: Urea was determined in serum according to the
methods of Tietz (1970) and Bonsnes. Triglyceride were determined
according to the methods of Stein and Myers (1995).
Alanine aminotransaminase (ALT) and Aspartate aminotransaminase (AST) activities
were determined according to the method of Reitman and Frankel
(1957). Glutathione-S-Transferase (GST) activity was determined according
to Habig et al. (1974). GST activity was defined
as the amount of enzyme catalyzing the formation of 1 mol of products per min
under condition of assay.
Activity = Å 340 nm/(9.6)x1000 = M/min
Statistical analysis: Mean and standard deviation of the obtained data
from each different experimental group were calculated and conducted according
the method described by Miller and Miller (1992).
Total antioxidant activity in Artemisia and Capparis extracts: Total antioxidants in Artemisia, Capparis, are shown in Table 1. The mean values were 675.33, 115.66 μmol Trolox per 100 mL for Artemisia, Capparis, respectively.
When total antioxidants are expressed as micromole Trolox per gram D.M, it was found that Artemisia contained higher antioxidants (135.66 ) when compared with Capparis (27.10) micromole Trolox per gram D.M.
Nutritional value of Capparis are presented in Table 2. As can be noticed from the Table 2 Capparis is a rich source of vitamin C. Each 100 g contained 4 mg of Ascorbic acid. It is also a rich source of iron and sodium.
|| Total antioxidants in Artemisia and Capparis
|| Nutritional value per 100 g of Capparis spinosa
|| Serum urea concentrations in control and in rats treated
with Artemisia monosperma and Capparis spinosa water extracts
|Means in the same column with different superscript are significantly
||Serum triglyceride concentrations in control rats and in rats
treated with Artemisia monosperma and Capparis spinosa water
|Means in the same column with different superscript are significantly
||Activities of AST and ALT in serum of control rats and in
rats treated Artemisia monosperma and Capparis spinosa water
|Means in the same column with different superscript are significantly
Serum urea: As shown in Table 3 there was a significant (p<0.05) increase in the concentration of urea in the positive control group. Administration of water extracts of Artemisia monosperma and Capparis spinosa resulted in a significant (p<0.05) decrease in levels of urea indicating usefulness of these medicinal plant extracts. Concentration of serum urea was 49.8 mg dL-1 in the control group (negative control) that increased to 73.10 dL in the positive control that received no plant extracts. Concentrations of serum urea decreased significantly (p<0.05) to 30.63 and 24.93 mg dL-1 after administration of Artemisia monosperma and Capparis spinosa, respectively.
Serum triglycerides: Serum triglycerides concentration increased significantly following lead acetate administration (Table 4). Triglycerides concentration in the negative control group was 74.63 mg dL-1 that increased to 178.66 mg dL-1 in the positive control. Treatment with water extracts from Artemisia monosperma and Capparis spinosa decreased the concentration of serum triglycerides to 106.33 and 87.20 mg dL-1, respectively.
As presented in Table 5. Lead acetate administration increased significantly (p<0.05) the activity of AST. The activity of this enzyme in the control group was 191.86 U L-1 that increased after lead administration to 219.80 U L-1 in the positive control group. This activity was significantly reduced to 187.59 and 169.94 U L-1 following administration of Artemisia monosperma and Capparis spinosa water extracts.
Activity of Glutathione-S-transferase in serum is shown in Table
6, the activity in serum of control rats, when expressed as Activity = A/9.6*1000
M min-1, was found to be 16.94 that decreased significantly (p<0.05)
to 11.25 in the positive control. When water extract of Artemisia monosperma
and Capparis spinosa was administered the activity of this enzyme increased
significantly (p<0.05) to 18.61and 21.35 A/9.6 1000 M min-1, respectively.
||Glutathione-S-transferase activity in serum of control rats
and in rats treated with Artemisia monosperma and Capparis spinosa
|Means in the same column with different superscript are significantly
In various cultures especially in the developing world, people are dependant on plants as medicine against various ailments. In Greek popular medicine a herbal tea made of caper root and young shoots is considered to be beneficial against rheumatism.
In the present study, it was found that lead ingestion disturbs many biological functions in rats mostly through free radicals generation, The antioxidant capacity in the body of rats as in other animals-is limited and usually exhausted by free radicals generated.
Lead administration increased the level of triglycerides and urea in blood.
Ingestion in drinking water of Artemisia monosperma and Capparis spinosa
water extracts lowered the concentrations of these metabolites. This may be
due to the presence of high levels of antioxidants in Artemisia monosperma
and Capparis spinosa (Table 1) and also due to high
level of vitamin C. Nutrient antioxidants may act together to reduce reactive
oxygen species levels more effectively than single dietary antioxidants, because
they can function as synergists (Eberhardt et al.,
2000; Rossetto et al., 2002; Podsedek,
The antioxidant activities of Artemisia species were investigated by
Juteau et al. (2002) and Kim
et al. (2003) and they found that extracts from Artemisia
caused significant increases in the rat liver cytosolic superoxide dismutase
(SOD) and catalase (CAT) that are highly involved in the antioxidant system
of the body. The increase in the activities of these enzymes will explain the
improvement in the pathological changes achieved in kidneys of lead intoxicated
rats. It was found that Artemisia extract significantly increased gastrointestinal
transit time and the reaction time to thermal stimuli, but had no effect on
the activity of alkaline phosphatase or concentrations of creatinine and urea
in plasma (Marrif et al., 1995). The plant extract
showed weak antimicrobial activity. On the other hand, Qureshi
et al. (1990) reported that ethanolic extracts of the aerial parts
of Artemisia abyssinica and A. inculta when subjected to acute toxicity
observations in mice for 24 h and chronic toxicity evaluation for 3 months.
It was found that external morphological changes, visceral toxicity, haematological
changes, spermatogenic dysfunction and effect on body weight and vital organ
weight were recorded. damage was observed in A. abyssinica treated mice
while A. inculta failed to produce any significant spermatotoxic effects.
Administration of lead acetate resulted in elevated activities of AST and ALT
enzymes that may be due to liver damage caused by the free radicals, however,
ingestion of Artemisia monosperma and Capparis spinosa water extracts
in drinking water decreased the activities of these enzymes indicating a protective
effect against liver damage. This is in agreement with the finding of Gadgoli
and Mishra (1999). They reported that p-Methoxy benzoic acid isolated from
the methanolic soluble fraction of the aqueous extract of Capparis spinosa
L. (Capparidaceae) was found to possess significant antihepatotoxic activity
against carbon tetrachloride and paracetamol induced hepatotoxicity in vivo
and thioacetamide and galactosamine induced hepatotoxicity in isolated rat hepatocytes,
using in vitro technique. through physicochemical and spectral studies.
Isolation, identification and antihepatotoxic activity of the compound is reported
for the first time in the plant.
On the other hand lead acetate intoxication reduced the activity of the enzyme
glutathione-S-transferase. Glutathione-S-transferase (GST) provides glutathione
(GSH) and helps to neutralize the toxic neutrophiles (Halliwell,
1994). Treatment with hot water extract of either Artemisia monosperma
and Capparis spinosa improved the activity of this enzyme leading to
improved formation of the very potent antioxidant, glutathione.
Previous phytochemical studies on Artemisa have yielded polyyne, sesquiterpene
(Stavri et al., 2005) and acetophenone natural
products (Bohlmann and Ehlers, 1977). Bohlmann
and Ehlers (1977) and known metabolites including the sesquiterpene spathulenol,
an hydroxylated polyyne and the flavonoid eriodyctiol-7-methyl ether. These
compounds were evaluated by those authors for their activity in 12-lipoxygenase,
antibacterial and found to protect against cytotoxicity. The mechanism(s) of
protection offered by Artemisia monosperma and Capparis spinosa
water extract are not clear at present and further work is needed.
It can be concluded that Artemisia and Capparis extracts contain high antioxidant compounds that protected rats against lipid peroxidation generated by lead. Isolation of individual compounds in these two plants is necessary for further studies.
This study was supported by a research grant from the Deanship of Scientific Research, Qassim University, Saudi Arabia. The author would like to thank Dr. Hassan M. Mousa and Dr. Ahmed Abdel-Salam from the Food Science and Human Nutrition Department, College of Agriculture and Veterinary Medicine for their valuable help with the Proximate and biochemical analysis and nutritive value.
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