Evaluation of
Anti-Inflammatory, Antipyretic and Antifungal Activity of Solanum laeve Dunal
P Muthumani*1,
R Meera1, Sweetlin1, P Devi1, B Kameswari1
and B Eswara Priya2
1K.M.
2Department of
Biotechnology,
ABSTRACT
Solanum leave Dunal var
biegninatum is small shrub is used by the tribals as a medicinal plant. The objective of the present
investigation was to study the anti inflammatory,
antipyretic and anti fungal activity potential in experimental animal models. The
isolated compounds obtained from the chromatography after the extraction
completed were used for (SA-I, SA-II, SA-III) these studies. Anti inflammatory
potential of isolated compounds (SA-I, SA-II, SA-III) was evaluated by
Carrageenan induced paw edema method in rats. Also antipyretic activity was
tested on animal models. The studies were conducted on wistar
rats of either sex (160-180g). The change in edema volume of the rat hind paw
was measured using plethysmometer. The SA-III of
isolated compound of Solanum leave dunal
inhibited the formation of edema to significant levels in rats treated with
carrageenan. At a dose of 100mg/kg orally, the SA-III produced 40.90%
inhibition in case of the carrageenan induced edema (p<0.01).
The isolated compounds
of Solanum leave Dunal
(SA-I, SA-II,SA-III) were also evaluated for
antipyretic activity on animals as per Vogel’s method. Solanum leave Dunal elicited a dose dependent
inhibition of rectal temperature compared with control group. The isolated
compound SA-II having maximum antipyretic activity which was compared with
standard inhibition at 300mg/kg p.o.
The fungal activity of
the isolated compounds (SA-I,SA-II,SA-III) were
prepared at various concentration of 1%,2% and 3% solution respectively. In
this the compound SA-II(C) having significant antifungal activity compared with
other compound against Candida albicans.
The results indicated
that the isolated compounds (SA-I,SA-II,SA-III) were
active against all the experimentally induced laboratory models of inflammation,
pyretic activities and also antifungal activity.
INTRODUCTION:
Most people in the
rural areas of the world depend largely on the herbs for the treatment of
several ailments because medicinal herbs constitute indispensable components of
tradition medicine practice due to low
cost, easy assess and ancestral experience1. Inflammatory responses
are mostly associated with pathological disorders 2. Although a good
number of plant species are used for this purpose, scientific and
pharmacological is scare or very little3. In recent years there has
been a growing interest to evaluate plants possessing antimicrobial activities
for various diseases4. A number of studies have been reported
dealing with antimicrobial screening of extracts of medicinal plants 5,6,7. Plant derived drugs have become a popular
alternative medicine in developing countries. Synthetic antifungal /
antibacterial drugs widely used at present are sometimes causing toxicity and
adverse drug reactions8. Further more, herbal medicines and
supplementation are considered less toxic than the synthetic compounds9.
However, folk medicines have not been studied extensively.
Preparation
of extracts (isolated compounds):
Although several
species of solaum are utilized for the cure of various
diseases.
Several workers reported this activity and the most notable report is activity
of solacasine form solanum psuedocapsicum against mycobactamium
smigmatitis Mitscher and
Wilson. Anti tumor activity of β Solamarine, and
Solapalmiotine isolated from S. tripartitum
against cells derived from the human carcinoma of nasopharynx
(Kupchan) against Sarcoma 180 in mice (cham) were reported Biological activities of α tomatine a glycol alkaloid like antifungal activity, anti
microbial and also anti diuretic was reported by Roddick
and concluded that activities were exhibited by partially purified methanolic extracts and not purified alkaloid. Roddick has also reviewed some enzymes inhibiting activity
of Solanum alkaloids and reported anti cholinesterase
activity 10-15
Materials and Methods:
Plant
material:
The plant has been identified in the Kodaikanal area and has been authenticated by Dr. V. Balasubramanian M.Sc. Phd.,
Taxonomist,
The plant was collected during months of
August-September which is the season for bearing the berries.
Since almost all the reports suggest berries
to be the main source of alkaloid. We have collected the plant during three
stages 1.Berries at green colour stage.2. Berries at greenish yellow 3. Berries at red colour (ripe).
Animals:
Wister rats were obtained from the
institutional animal house and they were kept in the departmental animal house
at 25±2°C and relative humidity 45-51.5%, light and dark cycles of 10 and 14h,
respectively for 1 week before and during the experiments for acclimatization.
The animals were provided with standard rodent pellet diet and water was
allowed ad libitum.
Rearing up of animals in the experimental span conformed to the norms of
institutional animal Ethical Committee (IAEC),
Rechromatography:
The fractions CDE (from chloroform: Ethylacetate
)and FG (from methanolic chloroform) found to
contain alkaloid as T.L.C. they were concentrated and pooled , and dissolved in
chloroform. They were rechromatographed on a smaller
column using methanol. The alkaloid elude from first four fractions (4x25ml).
The methanol was evaporated and alkaloid was crystalised
in chloroform in refrigerator for 2hours.
Table 1-
Antipyretic
activity of various extracts of Solanum leave Dunal
|
Drug |
Initial
temp. |
Temperature
in °C after hour shown (SEM ±) |
|||
|
0
hours |
1½
hours |
3hours |
4½hours |
||
|
Control |
37.6 |
39.2 |
39.3 |
39.5 |
39.5 |
|
Standard |
37.5 |
39.2 |
38.2 |
37.8 |
37.6 |
|
SA-I |
37.6 |
39.2 |
39.0 |
38.6 |
38.0 |
|
SA-II |
37.5 |
39.4 |
38.6 |
38.2 |
37.8 |
|
SA-III |
37.6 |
39.2 |
39.0 |
38.8 |
38.2 |
Table 2-
Anti-inflammatory
activity of various extracts of Solanum leave Dunal
|
Drug |
Dose mg/kg of body weight |
30min paw volume |
%decreaseof
paw volume |
60min paw volume |
%decrease of paw
volume |
120min paw
volume |
%decrease of paw
volume |
180 min paw volume |
%decrease of paw
volume |
|
Control |
Vehicle P.E.G.400 |
0.62 |
0.0 |
0.63 |
0.0 |
0.64 |
0.0 |
0.66 |
0.0 |
|
Standard drug[indomethacin] |
100mg/kg |
0.48* |
22.58 |
0.44** |
30.15 |
0.38** |
40.62 |
0.29* |
49.82 |
|
SA-I |
100mg/kg |
0.54** |
12.90 |
0.51* |
19.04 |
0.49* |
23.43 |
0.47* |
28.78 |
|
Sa-II |
100mg/kg |
0.60* |
3.22 |
0.53* |
15.87 |
0.47* |
26.56 |
0.42** |
36.36 |
|
SA-III |
100mg/kg |
0.53** |
14.51 |
0.47** |
25.39 |
0.43** |
32.81 |
0.39** |
40.90 |
*
Not
significant, ** Significant, Values
are mean ± SEM
Table 3- Anti fungal activity
of Solanum leave Dunal
|
Name of the compound |
Concentration of sample |
Diameter of zone of inhibition |
% of zone of inhibition |
|
SA –III Control |
1% 2% 3% 0.1ml |
- - 5mm - |
- - 29.4 - |
|
SA –I Control |
1% 2% 3% 0.1ml |
- - - - |
- - - - |
|
SA –II Control |
1% 2% 3% 0.1ml |
10mm 12mm 13mm - |
58.8 70.5 76.4 - |
|
Griseofulvin |
1% 2% 3% |
10mm 13mm 17mm |
58.8 76.4 100 |
The cholorform
extract on drying yielded white powdered with m.p203°C.This compound here after
referred SA-I.
Anti
inflammatory activity:16-20
Carrageenan
induced paw odema method:
The crude plant extracts were screened by
carrageenan induced rat paw edema method of Winter et al., the extracts, indomethacin standard drug and control vehicle
injected intraperitoneally,
30 minutes prior to the injection of carrageenan.
Albino rats of either ex weighing between
150-200gms were randomly distributed and were divided into to five groups, each
group consisting of four animals. The volume of paw of each animal was
determined before giving any drug. The first group served as control,(P.E.G.400) the second group served as standard, the third
group received SA-I, fourth and fifth groups received compounds SA-II,SAIII
respectively.
All the extracts were dissolved in P.E.G.400
and administered intra peritoneally of the dose of
100mg/kg of body weight of the rats. After 30minutes of duration 0.1 ml of 1%
W/V suspension of carrageenan was injected into the sub planter region of left
paw of each animal of the five groups.
The paw volume was measured at a time
interval of 30minutes, 60minutes, 120minutes and 180 minutes after the
administration of carrageenan. The degree of edema formation at the hind paw
volume was measure by plethsomometer.
The volume displacement has been expressed as
units are being equivalent to 0.72ml.
The percentage inhibition of odema has been calculated by the following formula and
results are tabulated and plotted.
Vc-Vt
--------- X 100
Vc
Where Vc
represents the average increase in paw volume of control and Vt represents the average increase in paw volume after the
administration of drug. Table 2
Antipyretic
activity21-23
Brewer’s
yeast method:
Antipyretic activity was conducted on Wistar strain albino healthy male rats weighing about
150-200gms. The rats showing 37.5±0.5°C were selected and then they were fasted
for 24 hours before inducing pyrexia. Their normal body temperature was
recorded. Pyrexia was induced by injecting subcutaneous by 12% w/v suspension
of yeast 1ml/200 body weight and they were allowed to feed. The animals were
divided into 5groups of 3 each and numbered.
After 10hours rectal temperature was recorded
using clinical thermometer introduced 2 cm into the rectum and keeping it
inside for one minute. The temperature recorded first after 10 hours of yeast
administration was taken as zero hour recording. The control, standard and the
compounds were given to the animals by asgric tube.
After the drug was administered, the temperature of all rats in each group were
recorded at an interval of 1½ hours,3 hours and 4½
hours. The mean temperature was found out for each group and compare with the
standard drug values.
The animals marked group-I received orally
1ml/200gm of body weight of 0.5% w/v solution of sodium laurylsulphate
and served as control.
The animals marked group –II received orally
300mg/kg body weight of acetyl salicyclic acid in
0.5% w/v suspension of sodium lauryl sulphate and served as standard the animals marked
group-III, IV and V received 300mg/kg of body weight of the various compounds.
The results are shown in the graph. Table 1
Antifungal
activity:24-31
The antifungal activity of
the compounds of Solanam leave Dunal
were evaluated by cup plate method with test organism pc Candida albicans. The powdered griseofulvin
(standard drug was dissolved in sterile water to produce a concentration of 1%,2% and 3% respectively. The solvent control was maintained
through out the experiment.
Preparation
of known concentration of compounds:
Like the standard drug solution, various
compounds of the plant were prepared at various concentration of 1%, 2% and 3%
solution respectively. The extracts were dissolved in an appropriate solvent.
From the solid culture the clinical sample of
Candida albicans
wee transferred into nutrient broth by using sterile inoculation loop the
nutrient broth was incubated for about 24 hours at 37°C. This showed sufficient
growth of fungi.
Cup-plate
or cylinder plate method:
For the screening of antifungal activity pour
plate method was used. Petridishes were filled to a
depth of 4-5mm with agar medium. Which has been previously
inoculated with the test organism. The temperature of the medium shold not exceed above 50°C when the organism were
inoculated. The petridishes were placed on horizontal
surface to ensure uniform thickness of the media and allowed to solidify.
After the agar has solidified 4 holes were
made in the medium with a sterile borer. About 0.1ml of 1%.2% and 3% solution
of extracts were added to the holes. The compounds were dissolved in the
appropriate solvent. A positive control and solvents were also added in the
same concentration to another bored petridishes and
zone of inhibition was measured after incubation at 37°C for 24hrs. the percentage of zone of inhibition was calculated by using
the formula. Table 3
Zone of inhibition of
test
Percentage Zone of inhibition =
---------------------------------- X 100
Zone of inhibition of Std.
Statistical
analysis:32
All the data were presented as mean ±SEM and
analyzed by Dunnett’s test and unpaired Students
t-test for the possible significant interrelation between the various groups. A
value of p<0.01 was considered statistically significant.
Results:
Carrageenan
induced paw oedema in rats:
The Solanum leave Dunal at the dose levels of 100mg/kg exhibited resistance
against carrageenan induced pain after 30min of the last dose and standard
drug. The average percent increase in paw volume indicates pain after treatment
was significantally(p<0.01) and dose dependently increased. The significant (p<0.01) effect of standard drug Indomethacin at the dose.
Anti
pyretic effect:
The result of the antipyretic study showed
the subcutaneous administration of the isolated compound SA-I (300mg/kg) caused
significant inhibition of the pyrexia induced by yeast. The antipyretic effects
of 300mg/kg extracts were comparable to that of acetyl salicylic acid b /w 30
and 60min after treatment.
Anti
fungal activity:
The compound SA-III(A), SA-I(B),SA-II(C) and
the standard drug also prepared various concentrations 1%,2% and 3% and the
activity was showed against candida albicans. The percentage of inhibition is significant
in SAII(C) as compared to standard drug Griseofulvin.
Discussion:
This study has investigated the scientific
reasons behind the folkloric use of Solanum leave Dunal, in the management of inflammatory conditions, anti
pyretic activity. The results indicated that the plant were active against all
the experimentally induced laboratory models of inflammation and pyrexia.
The isolated
compounds of Solanum leave dunal
inhibited the formation of paw oedema to significant
levels in rats treated either with Carrageenan
at a dose of 100 mg/kg orally, the SA-III of isolated compound of Solanum leave dunal
inhibited the formation of edema to significant levels in rats treated with
carrageenan. At a dose of 100mg/kg orally, the SA-III produced 40.90%
inhibition in case of the carrageenan induced edema (p<0.01). From that data
we can conclude that isolated compound SA-III of Solanum leave dunal inhibits the release of
serotonin and bradykinin (2h) and prostaglandin (3h)
.As the mechanism of carrageenan is it induces release of histamine (1h),
serotonin and bradykinin (2h), prostaglandin (3h).
Solanum leave Dunal elicited a dose
dependent inhibition of rectal temperature compared with control group. The
isolated compound SA-II having maximum antipyretic activity which was compared
with standard inhibition at 300mg/kg p.o.
The fungal activity of the isolated compounds
(SA-I, SA-II, SA-III) were prepared at various concentration of 1%,2% and 3% solution respectively. In this the compound
SA-II(C) having significant antifungal activity compared with other compound
against Candida albicans.
The results indicated that the isolated
compounds of Solanum leave dunal
were active against all the experimentally induced laboratory models of
inflammation, pyrexia and antifungal activities. The Anti inflammatory,
antipyretic and antimicrobial activities may be attributed to the presence of
alkaloids, carbohydrates, glycosides and steroids in the isolated compounds of Solanum leave Dunal32-35
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Received on 11.09.2009
Accepted on 29.11.2009
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Research Journal of Pharmacognosy and Phytochemistry. 2(1): Jan.-Feb. 2010, 25-29