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Determination of minerals and amino acids contents, anti-inflammatory and

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 World Academy of Science, Engineering and Technology 69 2012
Determination of Minerals and Amino Acids Contents, Anti-Inflammatory and Hepatoprotective
Effects of Syringa vulgaris L. Extracts
Mohammad Sanad Abu Darwish1*, Taha M. Rababah2, Bayan Abdulhaq4 , Kyslychenko V.S3., Andrey. I. Popik3., Korol V.V. 3, Ranya B. 2
Abstract— Leaves and flowers of Syringa Vulgaris L.(
Oleaceae ) are widely used in Ukrainian and Jordanian folk
medicine, for treatment of joints inflammation and to
extirpate kidney stones. Minerals and amino acids contents
were determined. The obtained results showed that, both
flowers and leaves of S. vulgaris contain 15 macro- and
microelements ,Si recorded the highest concentration.
Furthermore, up to 15 amino-acids were determined and a
high content of alanine, leucine, aspartic and glutamic acids
was found. Pertaining to the anti-inflammatory activity, the
intra-gastric administration of the fluid and thick extracts
at concentration 100 mg/kg exhibited the highest inhibition
activity against carrageenan and zymosan induced mice
paw oedema, whereas in the hepatoprotective assay the
fluid extract of S.vulgaris, at dose of 65 mg/kg exhibited 48,
17 % of hepatoprotective activity, while the thick extract of
S.vulgaris exhibited 52, 79 % hepatoprotective activity at
dose of 60 mg/kg.
Keywords— Amino acids, elements, Syringa vulgaris.
I. INTRODUCTION
S YRINGA
vulgaris L. (Oleaceae), Lilac is one of the most
common and most favourite ornamental plants in south eastern
europe. The leaves and flowers of S. vulgaris are widely used In
Ukrainian and Jordanian folk medicine. In the folk medicine of
Russia and Ukraine, the bark, leaves and flowers of s. Vulgaris
are used as a perfect anti-inflammatory remedy for treatment of
joints inflammations, rheumatism and rheumatoid arthritis [1],
while in Jordan the infusion and topical preparations of aerial
parts and seeds of S.vulgaris are used as Anti-helminthic, anti-
febrifuge remedies , and in the treatment of malaria [2].The
barks of S. vulgaris is the main source of the standard sample of
syringin (eleutheroside B), which exhibited a remarkable anti-
hypertensive activity [3]. The verbascoside extracted from
S.vulgaris exhibited antioxidant [4,5,6], and anti-inflammatory
activities[7,8]. Moreover, the tinctures of S. vulgaris flos f.
violácea and S. vulgaris flos f. alba exhibited antidiabetic
action and lowering serum total cholesterol, with ability to
stimulate antioxidant enzymes activity and reduce lipid per
oxidation. [9]. It was established that aqueous and ethanol
extracts of lilac spp. exhibited antiradical activities [1].
Mohammad S. Abu Darwish, Department of Basic and Applied
Sciences Shoubak University College, Al-Balqa Applied University,
Shoubak, 71911, Jordan.
Taha M. R., Ranya B. Esoh, Faculty of Agriculture, Jordan
University of Science and Technology, P.O. Box 3030, Irbid, 22110.
Kyslychenko V.S., Popik A.I., Korol V.V. ,Department of natural
compounds chemistry, National pharmaceutical university - Kharkiv,
Ukraine
Bayan Abdulhaq,Amman Arab University, Jordan.
Talib and Mahasneh, 2010[2] ,showed that the ethanol
extract of flowers and seeds of S. vulgaris , exhibited low
antiproliferative activity. Beside to the secondary metabolites,
the medicinal plants can contain primary constituents such as
amino acids and macro and micro-elements, which were
demonstrated to play an important biological role in human,
animal and plant health [10;11;10;and 12], and could cooperate
with other phytochemicals to accelerate the exertion of
efficacy. Many researches where conducted to evaluate amino
acids content in medicinal plants [13;14;15;16; 17 and18].
[14;15;16;7; and 18],while others were conducted to evaluate
the content of macro and micro-elemental composition
[19;20;21;22;23;24;25;26;12;27;28 and 29]. However some
researches evaluated the heavy metals content in Ornamental
plants [30; 31 and 32].
Data about amino acids and minerals constituents present in
leaves and flowers of S.vulgaris L. are still lacking. Therefore,
investigation and determination of these nutrients is important
to understand the therapeutic effects such as anti-inflammatory
and hepatoprotective activities. The aim of this work was to
study the quantitive and qualitative composition of amino acids
and macro and micro-elements in the leaves and flowers of S.
vulgaris, and to evaluate the anti-inflammatory and
hepatoprotective activities of its fluied and thick extracts, trying
to find a correlation between these compositions and their
pharmacological activities. This investigation will also
hopefully present new frontiers by improving the current
applications of this medicinal and ornamental plant and
provides a scientific basis for the economical uses of this plants.
II. MATERIALS AND METHODS
A. Medicinal Plants Samples
The Flowers and leaves of S. vulgaris L. were collected in
summer 2010 in Kharkiv, Ukraine. The collected Flowers and
leaves were air dried under shadow. A voucher specimen was
deposited at the Herbarium of Department of natural
compounds chemistry, National pharmaceutical university -
Kharkv, Ukraine. The dried samples of Flowers and leaves
were separately crushed into small pieces and sieved through
(0.5 mm) mesh sieve and kept dry for analysis.
1) Preparation of Fluid and Thick extract
The thick extract from the S.vulgaris leaves and flowers was
obtained by using 40% ethanol for plant material solvent in a
ratio of 1:5 maceration in correlation plant material – solvent
(1:5), 40 % ethanol was chosen as a solvent at 25 ºС during 24
hours. The received extracts were filtered, joined and
evaporated till the formation of a thick mass with humidity not
higher than 25 %.
3
The fluid extract from the S.vulgaris was obtained by
maceration in correlation plant material using a solvent 40 %
ethanol (1:10), was used as a solvent. The extraction was held
till the full exhaustion of the plant material at the temperature
of 25 ºС during 24 hours. The received extracts were filtered
joined and evaporated. Density was 0.907 g/cm3
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 World Academy of Science, Engineering and Technology 69 2012
B. Minerals Determination
1) Sample Preparation
The sample preparation started from careful carbonization of
the raw material by heating in the muffle furnace (the
temperature not higher than 500 ºC) with a preliminary sample
treatment by the diluted sulfuric acid. The sample evaporation
was held in the graphite electrode craters in the arc discharge of
alternating current (the spectra excitation source of the type
SES-28 in the current strength of 16 A and exposition of 60
sec).
2) Mineral Content Determination
The spectrograph DPhS-8 with a diffraction rating of 600
lines/mm and three-lenses system of the slit lighting was used
for the spectra obtaining and their registration. Using the
microphotometer MPh-1, the lines intensity in the spectra of
analyzed and calibration samples (CS) was measured. The
following conditions of the spectra photography were observed‫׃‬
the amperage of alternating current arc – 16 A; the arson phase
– 60 ºC; the frequency of ignition impulse – 100 discharges per
second; analytical gap – 2 mm; the width of the spectrograph
slit – 0,015 mm; exposition – 60 sec. The spectra were shot in
the range of 230-330 nm.
The photographic plates were shown, dried, then the
following lines were photometred in (nm) in the spectra of
analyzed and calibration samples and the background near
them. The differences between the line and background
blackening (S=Slf- Sm) in the spectra of analyzed (Sas) and
calibration (Scs) samples were calculated for each element after
the photometry results obtaining. The calibration graph was
built in the coordinates: the mean value of the difference
between the line and background (Scs) blackening – the
logarithm of element content in the calibration sample (lg C),
where C is expressed in percent to the basis. The content of
element in the plant material (a, %) was found on the graph.
The content of element in the plant material (x, %), was
а ⋅ m
calculated by the formula: х = , where m –
M Experimental animals were kept in the vivarium of the CRL
the mass of ash (g); M – the mass of the raw material (g); a –
the content of element in ash (%). The lower limits of the
adulteration content were taken into account that are: for Cu –
1·10-4 , Co, Cr, Mo, Mn, V – 2·10-4 , Ag, Ga, Ge, Ni, Pb, Sn, Ti
– 5.10-4; Sr, Zn – 1.10-2 %. The content of heavy metals was
determined by the method described in the European
Pharmacopoeia.
3) Test Solution
About 0.50 g of powdered plant raw materials were
separately and placed in a digestion flask. 6 ml of heavy metal-
free nitric acid R and 4 ml of heavy metal-free hydrochloric
acid R were added. The digestion flask was placed in the
microwave oven. The digestion was carried out in 3 steps
according to the following program, used for 7 flasks each
containing the test solution: 80 per cent power for 15 min, 100
per cent power for 5 min, 80 per cent power for 20 min. At the
end of the cycle, the flasks were cooled in air and to each 4 ml a
free of heavy metal sulphuric acid R was added. After cooling
of digestion flasks, the obtained clear and colorless solution
was introduced into a 50 ml volumetric flask. 1.0 ml of a 10 g/l
solution of magnesium nitrate R and 1.0 ml of a 100 g/l solution
1570
of ammonium dihydrogen phosphate R were added and diluted
to 50.0 ml with water R.
4) Blank Solution
6 ml of a free heavy metal- nitric acid R and 4 ml of a free
heavy metal hydrochloric acid R were mixed in a digestion
flask. The digestion was carried out in the same manner as for
the test solution.
The content of heavy metals was measured under
conditions described in the monograph 2.4.27., of the European
Pharmacopoeia.
5) Amino Acid Determination
Quantitative and qualitative amino acid analysis of the
common lilac flowers and leaves was carried out with the help
of amino acid analyzer T339M Mikrotechna-Praha. The
samples were dissolved in alcohol and placed in a flask of 50
ml, then concentrated hydrochloric acid was added, and
nitrogen was blown through to rewove air, the flask was then
hermetically closed by ground-in stopper and was put into a
thermostat with the heating temperature 120 ºC for 24 hours.
Each sample was filtered, into porcelain cup in which the
solution was softened by a nitrogen flow to remove
hydrochloric acid and to stabilize the pH solution within 1, 6
and 2.0 then that the sample was filtered once again through a
filter paper and caustic soda was added to the value of 2.2. To
the amino acid analyzer a 50 mcl of the sample was injected.
The quantitative analysis was carried out by comparing the
retention time of known standard amino acids with amino acids
in the sample. The quantitative content of amino acids (C, mcg)
in samples was calculated by the
С ⋅ S
formula: С = 1
, where C1 the
S 1
concentration of amino acid peak in the sample; S the area of an
amino acid peak in the sample; S1 the area of an amino acid
peak in the standard.
C.Anti-inflammentary and Hepatorotective Activities
(Central Research Laboratory) of the National University of
Pharmacy according to the standard sanitary norms on a
necessary diet. All studies were held according to the European
Council Directive 86/609/EEC of 24 November 1986 on the
approximation of laws, regulations and administrative
provisions of the Member States regarding the protection of
animals used for experimental and other scientific purposes.
1. Anti-inflammatory Activity
The study of anti-inflammatory activity of common lilac
extracts was carried out in two series. In the I series 45 white
mice weighing 20-22 g were used, in which aseptic
inflammation was caused by sub plantar injection of 0,05 ml of
1% λ-carrageenin solution (“Fluka”, Switzerland) into the right
hind paw. In the II series 45 white mice weighing 20-22 g were
also used, where exudative inflammation was caused by
zymosan (“Fluka”, Switzerland) which was used as a 0.05 ml
sub plantar injection in the form of 2% suspension. Fluid and
thick common lilac extracts in doses of 10, 25, 50 and 100
mg/kg each were used in both experimental series. All studied
objects were one-time administered intra-gastric 1 hour prior to
the induction of pathology. In 3 hours, the animals were
decapitated under the ether anesthesia and hinds were
amputated at the level of hip joint. Then the hinds were
 World Academy of Science, Engineering and Technology 69 2012

weighed and the absolute edema's value was calculated by the

difference between edematous and healthy foot. The anti-
inflammatory activity was valued in percents according to the
level of edema inhibition in animals that had administered the TABLE II
research objects in comparison to the animals from a control THE CONTENTS OF THE ELEMENTS (MG KG-1) IN THE LEAVES OF
pathology group[33]. SYRINGA VULGARIS L.
2. LD50 Content, № Content,
№ Element Element
The LD50 of the examined extracts was studied in 30 mice at mg kg-1 mg kg-1
1 Fe 1000 9 K 33300
one-time intragastrical administration with the doses interval 2 Si 4400 10 Ni 4.5
500 – 15000 mg/kg. For calculation of the median lethal dose 3 P 1700 11 Ca 10000
(LD50) the number of dead animals in each group was fixed and 4 Mn 1200 12 Mo <0.03
the LD50 value was determined using the conventional method. 5 Al 1300 13 Cu 20
3. Hepatoprotective Activity 6 Pb 3.0 14 Na 700
The hepatoprotective activity was studied on the model of 7 Ag <0.003 15 Zn 100
carbon tetrachloride hepatitis in mice (50% oil solution 10 8 Mg 5500 16 Sr 2
ml/kg intragastricaly). The studied extracts were administered 1
hour prior to and 2 hours after the injection of carbon
tetrachloride. The studied indexes (TBA-reactants in the liver Moreover, the low content of Pb in the studied flowers and
homogenate) were evaluated in 24 hours after carbon leaves of S.vulgaris could be due to, that their growth location,
tetrachloride injection. The weight liver coefficient and distanced from the main road and motor vehicles, which are
surviving of animals at using the studied extracts were also the leading factors of plant pollution with Pb
calculated. As experimental animals male mice weighing 20-22 [21,26,21,24and34] or due to that genetic differences between
g were used, that were divided into 4 experimental groups 15 plant species, play a role in the plant uptake of heavy metals
animals each: intact control; control pathology; animals that [28,26,28 and 34].
administered fluid extract at the dose of ED50; animals that The contents of Ni in leaves and flowers of S.vulgaris were
administered thick common lilac extract at the dose of ED50. also low and recorded 17 and 4.5 mg kg-1, respectively.
III MATH However, the detected amounts of Ni in both studied samples
The statistical analysis of the received data was performed not exceeded the toxic levels which are ranging from 10 to 100
using computer programs by variation statistic methods with the ppm [21]. This result could be due that the plant materials were
Fischer-Student reliability criterion. collected from natural habitat away from heavily traffic road. In
other hand the levels and limits of Ni are not yet been
I.V RESULTS AND DISCUSSION established by [35].
Results showed that the contents of Cu (30-20 mg kg-1) in
A. Determination of Micro and Macro Elements in the flowers and leaves of S.vulgaris were above the normal range
Flowers and Leaves (2-20 mg kg-1), respectively [24]. The content of Cu in studied
The results of macro and micro elemental content in flowers samples are less than founded in other medicinal plants, such as
and leaves of S.vulgaris are shown in Table 1 and 2. The Thymus vulgaris [28], Rosmarinus officinalis [21], Pergularia
contents of Pb which is known with high toxic effect to animal tomentosa and Cympobogon proximus [36] and various species
and plant health and can induce serious toxic effects in humans of Artemisia [20]. However, this variation could be due to
at low doses [34], were detected in flowers and leaves of genetic differences between plant species [20].
S.vulgaris in low amount 1.0 and 3.0 mg kg-1, respectively. The results of Zn concentration in flowers and leaves of
However, these contents of Pb are not exceeded the maximum S.vulgaris were 170 and 100 mg kg-1, respectively. The
level (10 mg kg-1) recommended by WHO in medicinal plants contents of Zn in the flowers and leaves of S.vulgaris were
[35]. within the typical amount of heavy metals in non polluted
TABLE I plants, which is ranging between 15 – 200 mg.kg-1 of dry
THE CONTENTS OF THE ELEMENTS (MG KG-1) sample [37].
IN THE FLOWERS OF SYRINGA VULGARIS L. Results in Table 1 and 2 showed that the concentration of Fe
Content, Content, was 1000 mg kg-1 in the leaves of S.vulgaris and decreased to
№ Element № Element
mg kg-1 mg kg-1 560 mg kg-1 in the flowers. This result is in agreement with El-
1 Fe 560 9 K 19400 Rjoob et al., 2008, [22] who reported that Fe was accumulated
2 Si 3300 10 Ni 17 in leaves, more than in flowers of Rosmarinus officinalis. These
3 P 940 11 Ca 4400 differences could be ascribed due to the plant foliar absorption
4 Mn 440 12 Mo 0.03
of Fe from the surroundings air [40]. However, the content of
5 Al 500 13 Cu 30
Fe in both studied samples did not reach it s typical amount
6 Pb 1.0 14 Na 330
7 Ag <0.003 15 Zn 170 [21].
8 Mg 1700 16 Sr 2 Silicon plays an important role in many body functions. It is
involved in the formation of bone and connective tissues and
could facilitate the formation of collagen components of the
bone matrix [38]. Moreover, Osório et al., 2006[39], suspected
that due to high content of Si in some medicinal plants, they
exhibited a wound healing efficacy. However, our results
showed that the contents of Si in Flowers (3300 mg kg-1) and

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 World Academy of Science, Engineering and Technology 69 2012

leaves (4400 mg kg-1) of S.vulgaris are higher than that found in known to improves insulin sensitivity, and reduce blood
some Russian medicinal plants whose total silicon content was pressure [44].
found between 7.4 g/kg and 35.9 g/kg [40] or than that found in Ca contents of studied samples were found in high
some Portuguese plants which were comprising between 1.25 concentrations, ranging from 4400 mg kg-1 in the flowers to
and 21.24 g/kg [39]. 10000 mg kg-1 in the leaves. however , our results revealed that
Results revealed that the content of Mn in the leaves of the studied S.vulgaris is a good source of Ca , which can
S.vulgaris is higher than in the flowers (1200 and 440 mg /kg, explain the using of the bark, leaves and flowers of S. Vulgaris
respectively).Since the critical threshold for Mn deficiency in in the folk medicine for treatment of joints inflammations, as
plant is < 10 ppm , it is clear that the leaves of S.vulgaris was well as Ca plays an important role in bones, teeth, muscles
more sufficiently provided with Mn , than the flowers. system and heart functions [45].
However, the contents of Mn in both studied organs of As shown in Tables 1 and 2, it is observed that Na
S.vulgaris were higher than the Permissible level, which was concentrations were differently accumulated by the tested
estimated as 200 ppm for plant [41] and higher than thus plants parts with the highest concentration found in the leaves
amounts obtained in another medicinal plants [19,41; 26; 12; (700 mg kg-1) and the lowest in the flowers (330 mg kg-
1
20and 36]. However, these results can suggest S.vulgaris to be ).However, the studied S.vulgaris accumulated Na less than
used for medicinal preparations to supplement Mn for various the species of Artemisia L. found in Pakistan [19].
body functions. B. Amino Acid
The concentrations of Sr in both studied samples were The obtained results (Table 3) revealed that leaves and
similar and equal 2 mg kg-1. This content is lower than thus flowers of S.vulgaris are containing fifteen amino acids. Nine
determined in some medicinal herbs consumed in Turkey which of them (Threonine, Valine, Methionine, Isoleucine, Leucine,
was ranged from 17.5 mg/kg in sage and 174 mg/kg in nettle. Phenylalanine, Histidne, Lysine, and Arginine) are belong to
The low content of Sr in our samples could be due to that the the essential amino acids group , while the six others Aspartic
root of S.vulgaris is not able to uptake it and is not able to acid, Serine, Glutamic acid, Glycine, Alanine, and Tyrosine are
continue the accumulation of Sr through over a period of time belong to the non-essential amino acids group. Moreover, the
[42]. contents of non-essential amino acids Aspartic acid, Serine,
The results for macro elements K, P, Al, Ag, Mg, Ca Glutamic acid, Glycine, Alanine, and Tyrosine are higher in the
and Na in leaves and flowers of S.vulgaris are shown in flowers of S.vulgaris than in it's leaves (1.38, 0.94, 0.78, 1.68,
Tables 1 and 2. The concentration of K was 19400 mg kg-1 in 0, 78 %, respectively).
the flowers and 33300 mg kg-1 in leaves. The tested organs of TABLE III
S.vulgaris gave evidence for high content of K and our results THE AMINO ACIDS COMPOSITION OF LEAVES AND
are in agreement with the earlier studies on various medicinal FLOWERS OF SYRINGA VULGARIS
plants [21; 27; 19 and 29]. However, this study showed that the Substance
Content, %
K content in leaves of S.vulgaris is higher than that found in the Leaves Flowers
flowers. This result is in agreement with Imelouane et al., *Aspartic acid
1 1.31 1.38
(Asp)
2011[29] who found that the content of K in the leafy organs is
*Threonine
higher than other nutrients as it is an activator of some 2
(Thr)
0.97 0.83
enzymes. 3 *Serine (Ser) 0.89 0.94
P contents in the flowers and leaves were also determined *Glutamic acid
4 2.38 2.70
and found to be high (940 and 1700 mg kg-1) respectively, (Glu)
which gives indication of satisfactory level of P supply in 5 *Glycine (Gly) 0.65 0.78
S.vulgaris. However, the content of P in our investigation is 6 *Alanine (Ala) 1.63 1.68
higher than that found in some other medicinal plants [29]. 7 *Valine (Val) 0.,84 0.87
Al contents in our samples of S.vulgaris varied from 500 mg *Methionine
8 0.39 0.52
(Met)
kg-1 in the flowers and 1300 mg kg-1 in the leaves. However,
9 *Isoleucine (Ile) 0.78 0.84
Lo´pez et al. 2000[43] found that the Al content in 72 samples 10 *Leucine (Leu) 1.37 1.24
of 17 different spices and aromatic herbs ranged from 5.20 to 11 *Tyrosine (Tyr) 0.59 0.78
35.30 mg/kg, while Imelouane et al., 2011[29] showed that the *Phenylalanine
12 0.75 0.82
content of Al ranged from 12265 mg/kg in Wormwood and (Phe)
79152 mg /kg in Thyme. 13 *Histidne (His) 0.49 0.49
The concentration of Mg ranged from 1700 mg kg-1 in the 14 Lysine (Lys) 0.72 0.81
flowers to 5500 mg kg-1 in the leaves of S.vulgaris. Higher 15 *Arginine (Arg) 0.94 0.97
values of Mg were reported by Imelouane et al., 2011[29]
(30914 mg /kg in Wormwood to 68382 mg/kg in Rosemary). Among all of detected non-essential amino acids, Glutamic
Achak et al., 2009[20], determined the Mg content of Juniperus acid recorded the highest content in leaves and flowers of
thurifera L var. africana, J. phoenicea and J. oxycedrus with a S.vulgaris (2,38 and 2.70%, respectively), while Glycine
range from 27,6 to 61,7 mg/1000mg.This variation could be recorded the lowest (0,65 and 0,78 % , respectively).
due to the content of Mg in soil , as well as level of Mg in
plants depends to a large extent on soil type [29]. However , the
existing anti-diabetic action and lowering serum total The content of detected essential amino acids in leaves and
cholesterol of tinctures of Syringae vulgaris flos f. violácea and flowers of S.vulgaris are various. However, the concentrations
Syringae vulgaris flos f. Alba founded by Berbecaru- of detected essential amino acids in flowers of S.vulgaris,
Iovan,2009[9], could be due to the content of Mg which is well except Threonine and Leucine, were higher than that recorded

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 World Academy of Science, Engineering and Technology 69 2012

in the leaves. Among all detected essential amino acids Leucine

recorded the highest content in leaves 1,37 % and 1,24 % in the
flowers, while Histidne recorded the lowest content (0,49%) in
both leaves and flowers of S.vulgaris.
TABLE V
However , the results revealed that the leaves and flowers of THE ANTI-INFLAMMATORY ACTIVITY OF COMMON LILAC EXTRACTS
S.vulgaris are a good source of amino acids which are play ON THE ZYMOSAN OEDEMA MODEL IN MICE
important roles , as building unites of proteins and as Anti-
intermediates in metabolism , which also can explain the Experiment Oedema
№ n inflammatory
medicinal and biological value of S.vulgaris , where many conditions weight, mg
activity, %
researches indicated the important role of amino acids in 1 Control
5 0.192±0,01 -
immune function [46,47and48] and as antioxidant agents [48; pathology
49] 2 Fluid
C. Anti-Inflamantary extract, 10 5 0.133±0,00 30.73
mg/kg
The Results of anti-inflammatory activity of fluid and thick
3 Fluid
extracts of S.vulgaris on the carrageen in oedema and zymosan extract, 25 5 0.124±0,00 35.42
oedema models in mice are shown in Table 4 and 5. mg/kg
4 Fluid
TABLE IV extract, 50 5 0.113±0,00 41.15
THE ANTI-INFLAMMATORY ACTIVITY OF COMMON LILAC EXTRACTS mg/kg
ON THE CARRAGEENIN OEDEMA MODEL IN MICE 5 Fluid
Anti- extract, 100 5 0.106±0.00 44.79
Experiment Oedema mg/kg
№ n inflammatory
conditions weight, mg 6 Thick
activity, %
1 Control extract, 10 5 0.146±0.01 23.96
5 0.172±0.011 - mg/kg
pathology
2 Fluid extract, 7 Thick
5 0.113±0.005 34.30 extract, 5 0.129±0.,01 32.81
10 mg/kg
3 Fluid extract, 25mg/kg
5 0.103±0.006 40.12 8 Thick
25 mg/kg
4 Fluid extract, extract, 50 5 0.117±0.01 39.06
5 0.096±0.013 44.19 mg/kg
50 mg/kg
5 Fluid extract, 9 Thick
5 0.089±0.007 48.26 extract, 100 5 0.111±0.01 42.19
100 mg/kg
6 Thick mg/kg
extract, 10 5 0.123±0.008 28.49
mg/kg The results observed in rat bow oedema assay reveled a
7 Thick significant inhibitory activity in carrageenin and zymosan
extract, 25 5 0.110±0.005 36.05
induced paw inflammation. The present results showed anti-
mg/kg
8 Thick
inflammatory activity of both liquid and fluid extracts of
extract, 50 5 0.099±0.009 42.44 S.vulgaris increasing proportionally with increasing the
mg/kg concentration of examined extract. The 100 mg/kg
9 Thick concentration of fluid and thick extracts of S.vulgaris inhibited
extract, 100 5 0.094±0.010 45.35 the carrageenin induced paw inflammation in 48,26 and
mg/kg 45,35%, respectively . While the same concentration of both
tested extracts inhibited the zymosan induced paw
inflammation in 44,79 and 42,19 %,respectively. However, the
fluid extracts exhibited a stronger anti-inflammatory activity in
carrageenin and zymosan induced paw inflammation more than
thick extracts. The median effective doses of the studied
extracts were calculated using the method of least squares.
Thus, in carrageenin paw oedema the ED50 of the fluid and
thick extracts was evaluated to be 67,00 mg/kg,and 62,60
mg/kg , respectively. While in zymosan was evaluated to be to
be 62,40 mg/kg and 56,00 mg/kg, respectively . Results are in
agreement with [1] who showed that the S.vulgaris extracts
exhibited anti-inflammatory activity.
Moreover, the obtained toxicological parameters showed
that no lethal cases were been detected at intragastric
administration of S.vulgaris fluid extract. Supposing the LD50
of the studied extract to be higher than 15000 mg/kg. While at
intragastric administration of S.vulgaris thick extract, only one
lethal case was detected at the dose of 15000 mg/kg. These

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 World Academy of Science, Engineering and Technology 69 2012

results can suppose that the LD50 of the S.vulgaris thick extract animals. Also, threonine prevented apoptosis, stimulated cell
to be 5000 mg/kg. growth and promoted antibody production in lymphocytes [53,
The anti-inflammatory activity of S.vulgaris extracts 48].
could be explained due to their active ingredients such as Unnikrishnan and Rao1990[62] found that methionine had
verbascosides [50,7] and/or their content of amino acids. good anti-inflammatory activity in carrageenan induced paw
Verbascosides isolated from S.vulgaris and chemically belong edema .Farshid et al .,2010[63] found that Histidne induced
to phenylpropanoids a class of plant-derived organic anti-inflammatory activity by reducing paw edema and
compounds that are biosynthesized from the amino acid neutrophile infiltrationin induced by carrageenan. The
phenylalanine, exhibited remarkable anti-inflammatory activity antiinflammatory activity of Aspartic acid in the carrageenin
in a model of inflammatory bowel disease in mice [7] and induced paw inflammation has been reported by Naik and
exerted an anti-inflammatory role during experimental Sheth, 1978[56]. Histidine and some amino acids such as
periodontitis [8].Moreover L-beta-phenylalanine inhibited cysteine, and glycine exhibited anti-inflammatory effects
carrageenan-induced oedema [47] during endothelial inflammation [72]. Farshid et al.,2011[63]
Many carried researches provided that amino acids improve showed that histidine potentiated the anti-inflammatory effect
the immune functions of living organism [48] and exhibited a of dexamethasone in histamine-induced local inflammation.
significant anti-inflammatory activity, where Tryptophan, 3.4. Hepatoprotective Activity
phenylalanine, alanine, cystine, hydroxyproline, and tyrosine The hepatoprotective activities of the fluid and thick extracts
significantly decreased the gelatin-induced abdominal of S.vulgaris on the model of carbon tetrachloride hepatitis in
inflammation in mice [46]. Jain and Khanna., 1984 [51] found mice are shown in Table 6. The obtained results revealed that
that L-glutamine, has marked anti-inflammatory activity, and the both studied extracts possess expressed hepatoprotective
this activity due that L-glutamine partially mediate its anti- activity as evidenced by the increase of the number of survived
inflammatory activity by interfering with the action and/or animals, increase of the weight liver coefficient, normalization
synthesis of prostaglandins. Alanine , is responsible to Inhibit of TBA-active products in the organ’s homogenate and high
the apoptosis and stimulate of lymphocyte proliferation[48]. antioxidant activity. The fluid extract of S.vulgaris, at dose of
Alanine could influence immune function, due that it is the 65 mg/kg exhibited 48, 17 % of hepatoprotective activity, while
major substrate for the hepatic synthesis of glucose, a the thick extract of S.vulgaris exhibited 52,79 %
significant energy substrate for leucocytes [52]. Duval et al. hepatoprotective activity at dose of 60 mg/kg. The expressed
1991[53] and Franek & Sramkova, 1996[54], showed that hepatoprotective activity of S.vulgaris extracts could be due to
Serine prevented apoptosis, stimulated cell growth and their active ingredients such as verbascosides. This result is in
increased antibody production in lymphocytes. agreement with Lee et al. 2004 [65], who showed that
Glycine participates in the synthesis of purine nucleotides, verbascoside exhibited in vivo hepatoprotective effect against
glutathione and haem [55]. Morever Konashi et al. 2000[62] CCl4 in mice at relatively low dose. In other hand, many
showed that Glycine reduces inflammatory reactions and researchers reported that some amino acids exhibited
morbidity in pathogen-infected animals. During endothelial hepatoprotective activity [66]. Lee and Kim, 2007[65]
inflammation, Glycine exhibited antiinflammatory activity in reported that β-alanine showed hepatoprotective effect against
human coronary arterial endothelial cells. [56]. CCl4-induced liver injury. Administration of a mixture of
Tyrosine, is the immediate precursor for the synthesis of leucine and zinc sulfate (4: 1; 100 mg/kg body weight) induced
dopamine and melanin [55], which are reduce the synthesis of a hepatoprotective effect avoiding the ultrastructural injury of
pro inflammatory cytokines, induce the production of anti- hepatic tissue and the disturbance of free amino acid
inflammatory mediators, and regulate lymphocyte proliferation, metabolism [67].
platelet aggregation and the phagocytic activity of neutrophils Since silica was the major microelement founded in both
[57, 58]. studied leaves and flowers, we might assume that
The essential amino acid Leucine plays role in Regulation hepatoprotective activity shown in our assays could be
of immune responses [51]. However, Konashi et al. 2000[40] due to this element, where Hsu et al., 2010[68] showed
found that leucine appears to exert a greater effect on immune that silica exhibited potent hepatoprotective activity on
function than isoleucine and valine. Saxena et al.,1984 [47]
CCl (4)-induced liver damage in mice.Although
showed that L-isoleucine, DL-isoleucine and L-leucine
synergistic hepatoprotective activity effect of silicon,
exhibited anti-inflammatory activity in many test models of
inflammation except formaldehyde-induced inflammation.
amino acids and author major compounds of S.Vulgaris
In subjects with inflammation, plasma concentrations of such as verbascoside cannot be discarded.
TABLE VIII
arginine decrease markedly [59].Moreover, Field et al. THE HEPATOPROTECTIVE ACTIVITY OF COMMON LILAC EXTRACTS ON
2000[60] found that dietary arginine supplementation enhances THE MODEL OF CARBON TETRACHLORIDE HEPATITIS IN MICE
immune function in various models of immunological disorders TBC- Hepato-
. Further, dietary supplementation with 0·83% arginine Survi Weight
Object of reactants, protective
№ n val, coeffici
enhanced the immune status of pregnant sows and neonatal study, dose
% ent
ng/g of activity,
pigs, thereby reducing morbidity and mortality in response to tissue %
infectious pathogens [55]. Field et al. 2002[60] and Suchner et 1 Intact 4.18± 2,37±
15 100 -
control 0.13 0,16
al. 2002[61] found that enteral or parenteral provision of 2 Control 2.75± 5,19±
arginine improves immune functions and clinical outcomes in 15 0 -
pathology 0.08 0,22
patients with, cancer, HIV infection, and major traumas. 3 Fluid
3.79± 2.69±
Kim et al. 2007[55] showed that threonine is a major extract, 65 15 80 48.17
0.05 0.11
component of intestinal mucin and Plasma -globulin in mg/kg

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 World Academy of Science, Engineering and Technology 69 2012

4 Thick basis of hydrophilic compounds of Eucalyptus leaves," Herba

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