Journal of Applied Pharmaceutical Science 01 (10); 2011: 98-101

ISSN: 2231-3354
Received on: 22-11-2011
The analgesic and anti-inflammatory activities
Revised on: 28:11:2011
Accepted on: 09-12-2011
of the hydroalcholic extract of “Shirishadi
compound” in animal model

Divya Kajaria, J.S. Tripathi , S.K.Tiwari and B.L. Pandey

ABSTRACT

The main objective of the present investigation is to evaluate the anti-inflammatory &
analgesic activity of ethanolic extract of Shirishadi polyherbal compound on rats. Shirishadi
Divya Kajaria, J. S. Tripathi ,
compound consist of three herbal drugs namely- Shirisha (Albizzia lebbeck), Nagarmotha
S.K.Tiwari
Department of Kayachikitsa,
(Cyprus rotandus) & Kantakari (Solanum xanthocarpum).In Ayurveda (ancient Indian system of
Faculty of Ayurveda, I.M.S., medicine) all these herbs alone or in combination with other herbs are commonly used in the
Banaras Hindu University, Varanasi, managmant of bronchial asthma. In the carrageenan-induced rat paw edema test for acute
India. inflammation, the extract of Shirishadi compound in doses of 50mg, 200 mg and 500 mg/kg
body weight showed 77% and 79% and 81% inhibition of edema, respectively, at the end of 4h
which is comparable to that of standard ( endomethacin) i.e. 92%. In the acetic acid induced
writhing test the extract of Shirishadi compound ( 200 and 500 mg/kg body weight) showed a
significant (p<0.001) reduction in the number of writhes with 65.6% and 70.9% of inhibition,
B.L. Pandey
respectively. In radiant heat tail-flick test the crude extract produced 58.1% (p<0.001) and 61.1%
Depatment of Pharmacology,
I.M.S., Banaras Hindu University,
(p<0.001) elongation of tail flicking time 30 minutes after oral doses of 200 and 500 mg/kg body
Varanasi, India. weight respectively . After 60 minutes the extract showed 56.3% (p<0.001) and 59% (p<0.001)
elongation of tail flicking time. Experimental results showed that Shirishadi compound has
persuasive anti-inflammmatory property along with significant analgesic activity.

Keywords: Shirisadi polyherbal compound, Anti-inflammatory activity, Analgesic activity,

Radiant heat tail-flick test, Writhing test.

INTRODUCTION

Asthma is a common disease that is rising in prevalence worldwide with the highest
prevalence in industrialized countries. Asthma affects about 300 million people worldwide and it
has been estimated that a further 100 million will be affected by 2025(1-2). Asthma is defined as
disorder characterized by chronic airway inflammation and increased airway responsiveness
resulting in symptoms of wheezing, cough, chest tightness, and dyspnea. It is characterized
functionally by the presence of airflow obstruction which is variable over short period of time or is
reversible with treatment. It is not a uniform disease but rather a dynamic clinical syndrome which
has a number of clinical patterns. Current asthma therapy lack satisfactory success due to adverse
For Correspondence effect, hence patients are seeking complementary and alternative medicine to treat their asthma.
Prof S.K.Tiwari
Medicinal plant used for the treatment of asthma should have anti-inflammatory,
M.D. Ph.D
Professor & Head of Department immunomodulatory,antihistaminic, smooth-muscle relaxants and allergic activity. The basic
Institute of Medical Sciences, pathology of Asthma starts with the process of inflammation so to show the antiasthmatic activity
Faculty of Ayurveda,
Depatment of Kayachikitsa,
of drug first step involve to demonstrate the anti- inflammatory activity of drug. Inflammation is a
Banaras Hindu University, Varanasi, defensive and normal response of the body to any noxious stimulus, varying from acute, transient
India. and highly localized response to a minor mechanical injury to a complex sustained response
 Journal of Applied Pharmaceutical Science 01 (10); 2011: 98-101

involving the whole organism. It has been stressed that reaction in a number of animal species. Winter et al. (1962)
inflammation is a process and not a state (Florey, 1970). Acute introduce the carrageenin edema of rat hind paw for assay of anti-
inflammation refers to a response with a rapid onset and relatively inflammatory drugs. The reproducibility of the fact that
short duration and characterised by particular vascular inflammation entirely depends upon local inflammatory reaction
phenomenon. Albezzia lebbeck, Cyperus rotandus and Solanum devoid of anigenic properties, has made carrageenin the most
xanthocarpum used in the present research trial are use from era widely employed phlogistic agent. For the present experiment,
to treat asthma in Ayurveda. In order to search the probable carrageenin suspension was prepared as a homogenous suspension
mechanism of their action and their pharmacological activity the of powder in 0.9% sodium chloride solution (sterile normal saline)
present research trial was conducted with the aim to show the anti- with the help of mortar & pestel. A volume of 0.1ml of suspension
inflammatory and analgesic activity of drug in animal model. was injected through a 26 gauge needle into the plantar surface of
the right hind paw below the plantar aponeurosis 1h after the oral
MATERIALS AND METHODS administration of test materials. The volume of hind paw of the
rats upto the ankle joint was measured plethysmographically, by
Plant collection
the mercury displacement method. The volume was measured 1h,
The plants Albizzia lebbeck, Cyprus rotandus and
2h ,3h, 4h & 24 after the administeration of drug. The extract was
Solanum xanthocarpum were collected from local market of
administered at 50, 200 and 500 mg/kg body weight.
Varanasi. The identification of the drugs was done by Prof.A.K.
Endomethacin 25 mg/kg body weight was used as standard anti-
Singh, Department of Dravyaguna, S.S.U., Varanasi.
inflammatory agent.
Extraction of the plant material and sample preparation
Acetic acid induced writhing test
Hydroalcoholic Extraction ( Distilled water: Ethanol =
The peripheral analgesic activity of hydroalcholic extract
2:1) of drug was carried out by Hot percolation method through
of Shishadi Ayurvedic compound was measured by the acetic acid
soxhlet appratus. Thereafter extract was dried using rotary
induced writhing test as described earlier (Saha et al., 2007).
evaporator and dried extract was put to the process of
Briefly, the inhibition of writhing produced by the plant extract
standradization.The extract was dissolved in distilled water and
was determined by comparing with the inhibition produced by the
three different concentration namely 50mg/ml, 200mg/ml &
control group. Aminopyrine at oral dose of 50 mg/kg was used as
500mg/ml was prepared.
standard analgesic agent. Intraperitoneal injection of acetic acid
(0.7%) at a dose of 0.1 ml/10g of body weight was used to create
Drugs and Chemicals
pain sensation. The number of writhings was calculated for 10 min,
Aminopyrine, Carrageenan, Pentazocin, Endomethcin &
10 min after the application of acetic acid.
Acetic acid were purchused from Sigma-Aldrich, Germany.
Radiant heat tail-flick method
Experimental animal
The central analgesic activity of the plant material was
Adult Charles Foster Albino rats (150+ 30g) of either
studied by measuring drug-induced changes in the sensitivity of the
sex were obtained from the Animal Research Branch of the
pre-screened (reaction time: 2-4 sec) mice to heat stress applied to
Institute of Medical Sciences, Banaras Hindu University, Varanasi.
their tails by using a Medicraft Analgesiometer Mask-N (D’Amour
The animals were housed in polyvinyl cages and. were fed on
and Smith,1941) and described previously (Saha et al., 2007).
commercial pellet diet (Amrut, Pranav Agro Industries Ltd, India).
Briefly, the current intensity passing through the naked nicrome
They were group housed under standard conditions of temperature
wire was maintained at 5 ampere. The distance between the heat
(22 ± 20C), relative humidity (60 ± 5%) and 12:12 light/dark cycle,
source and the tail skin was 1.5 cm and cut-off reaction time was
where lights on at 0700 and off at 1900 h). The saline fed group
fixed at 10 second to avoid any tissue damage. Morphine was used
served as control and one group was treated with a standard drug in
to compare the analgesic effect of the plant extract.
each protocol. Before experimentation, the animals were kept on
fast for 24 h but water was given ad libitum except during Table 1: Anti-inflammatory activity of crude extract of Shirishadi compound by
experimental test period. During experiments, animals were also carrageenan induced rat paw edema.
observed for any alteration in their general behavior. % Increase in Paw Volumes (ml × 1000) ± SEM (%
All the experiments and the care of the laboratory animals GROUP inhibition)
1h 2h 3h 4h 24h
were according to current ethical guidelines by the Committee for Control 1.78 + 0.77 3.0 +0.15 3.61 + 0.20 4.0± 1.2 1.93 ± 0.11
the Purpose of Control and Supervision on Experiments on Standard 0.72 ± 3.7 0.69 ± 0.60 0.67 ± 0.66 0.65 ±0.37 0.66 ± 0.52
Animals (CPCSEA), Ministry of Environment and Forests, (Indomethacin (59%) (77%) (81%) (92%) (66%)
25mg/kg)
Government of India, New Delhi. Shirishadi 1.06 ± 0.68 0.99 ± 0.66 0.93 ± 0.33 0.89 ±0.33 0.78 ± 0.41
50mg/ Kg bwt (40%) (67%) (74%) (77%) (59%)
Shirishadi 1.0 ± 1.0 0.97 ± 0.14 0.92 ± 0.24 0.88 ±0.57 0.77 ± 0.24
Anti-inflammatory study 200mg/Kg bwt (43%) (67.66%) (74.28%) (79%) (60%)
Carrageenin,a sulphated polysaccharide,extracted from Shirishadi 0.95 ± 0.33 0.95± 0.37 0.83 ± 0.66 0.76 ± 0.63 0.66 ± 0.33
500mg/Kg bwt (46.6%) (68%) (77%) (81%) (65%)
sea weed, has been extensivelyused to induce inflammatory
 Journal of Applied Pharmaceutical Science 01 (10); 2011: 98-101

Values are mean+SEM (N=3). Data were analyzed by one-way ANOVA followed by Dunnet’s
Table :2 Effect of Shishadi Extract on acetic acid induced writhing response in test and P values <0.05 were considered statistically significant.
rodents.
Groups Dose of Drug Writhingb %
Inhibition RESULTS AND DISCUSSION
Control -- 15.6 ± 0.50 --
Standard Pentazocin 4.3 ± 0.66 ** 71.2% In the carrageenan-induced rat paw edema test (table 1)for acute
10mg/Kg, i.p. inflammation, the extract of Shirishadi compound in doses of 50mg, 200
Aspirin 25 8.0 ± 1.15 ** 63.5% mg and 500 mg/kg body weight showed 77% and 79% and 81%
mg/Kg, i.p.
200mg/ Kg bwt, 7.66 ± 0.88 ** 65.6%
inhibition of edema, respectively, at the end of 4h which is comparable to
Shirishadi p.o. that of standard ( endomethacin) i.e. 92%. In the acetic acid induced
500 mg/ Kg bwt, 6.33 ± 0.88 ** 70.9% writhing test the extract of Shirishadi compound ( 200 and 500 mg/kg
p.o.
body weight) showed a significant (p<0.001) reduction in the number of
F 46.94 -- writhes with 65.6% and 70.9% of inhibition, respectively (table2). In
One way df 4,12 -- radiant heat tail-flick test the crude extract produced 40.74% (p<0.001)
ANNOVA and 61.48% (p<0.001) elongation of tail flicking time 30 minutes after oral
P <0.001 -- doses of 200 and 500 mg/kg body weight respectively (table 3). After 60
a
minutes the extract showed 31.29% (p<0.001) and 41.37% (p<0.001)
1hr after treatment, mice were injected i.p. with 0.7%(v/v) acetic acid (0.1ml/10g);
elongation of tail flicking time. The constriction response of abdomen
10 minutes after the injection, the number writhing was counted for 10 min. b
Values are mean ± SEM (n = 5, for control & 3in drug treated groups); One-way produced by acetic acid is a sensitive procedure for peripheral analgesic
ANOVA; **P<0.001, compared to control. agents. This response is believed to be mediated by the prostaglandin
pathways (Ronaldo et al., 2000). The extract of Shirishadi compound
Table 3: Effects of crude extracta on radiant heat tail-flick produced antinociceptive activity and thus indicates the presence of
response in rodents. analgesic components that might influence the prostaglandin pathways. In
Reaction Time (sec) the radiant heat tail-flick test, the polyhderbal extract prolonged the stress
Groups Dose of 30 (mins) 60 120 (mins) tolerance capacity of the rat, indicating the possible involvement of a
Drug (% elongation) (mins) (%
higher center (Whittle, 1964). Carrageenan-induced inflammation in the
( % elongation)
elongation)
rat paw represents a classical model of edema formation and hyperalgesia,
which has been extensively used in the development of nonsteroidal anti-
Control -- 4.65 ± 4.82 ± 4.98± inflammatory drugs and selective COX1-2 inhibitors (Winter CA,et.al
0.15 0.16 0.20 1962).. Several lines of evidence indicate that the COX-2-mediated
increase in prostaglandin (PG) E2 production in the central nervous system
Standard Pentazocin 8.65 ± 6.45 ± 5.89 ±
10mg/ kg 0.71** 0.45** 0.37** (CNS) contributes to the severity of the inflammatory and pain responses
bwt, i.p. in this model. In the paw, the early phase was associated with increases in
200 mg/ Kg PGE2 and thromboxane (TX)B2 levels and with a peak of COX-2 (Vinegar
Shirishadi bwt, p.o. 7.41 ± 7.10 ± 6.08 ± R,et.al.1969). Therefore, the inhibition of carrageenan induced
Compound 1.17** 0.30** 0.21**
(58.1%) (56.3%) (49%)
inflammation by the extract of Shirishadi compound could be due to the
500 mg/ inhibition of the enzyme cyclooxygenase and subsequent inhibition of
Kg bwt, 7.98 ± 7. 57 ± 6.75 ± prostaglandin synthesis. The present study on extract of Shirishadi
p.o. 0.12** 0.45** 0.15** compound has demonstrated that this compound has significant analgesic
(61.1%) (59%) (54%)
and anti-inflammatory properties, and thus can be use in Bronchial asthma
F 68.5 27 5.34 and other inflammatory & painful conditions.
One way
ANOVA P < 0.001 < 0.001 < 0.001 REFERENCES
df 7,40 7, 40 7,40 Chintawar SD, Somani RS, Kasture VS and Kasture SB.
a .per oral administration of vehicle and crude extract, radiant heat intensity was 6 Nootropic activity of Albizia lebbeck in mice. J. Ethnopharmacol.
c
amp. b morphine was administered sub-cutaneously. Values are mean ± SEM (n 2002;81(3): 299-305.
=5 ); One-way ANOVA; df = 7, 40; **P<0.01, *P<0.05 compared to control. Crunkhorn P and Meacock SC. Mediators of the inflammation
induced in the rat paw by carrageenin. Br.J. Pharmacol. 1971;42: 392-
Table 4 Intergroup comparision of shirishadi treated and Endomethacin treated
402.
group with control by using One Way ANNOVA followed by Post- Hoc test.
D’Amour FE and Smith DL. A method for determining loss of
Change in Paw Volume expessed as Mean ± SE ( per hour) pain sensation. J. Pharmacol. Exp.Ther., 1941;72: 74-79.
Groups 1h 2h 3h 4h 24h Gupta RS, Kachhawa JB and Chaudhary R. Antifertility effects
Shirishadi 1.06 ± 0.99± 0.93 ± 0.89± 0.78 ± of methanolic pod extract of Albizia lebbeck Benth. in male rats. Asian J.
50mg/ 100g bwt 0.068** 0.006** 0.03** 0.003** 0.04**
Shirishadi 1.10 ± 0.97 ± 0.93 ± 0.89± 0.77 ± Androl. 2004; 6(2): 155-159.
200mg/100g bwt 0.05** 0.01** 0.02** 0.05** 0.02** Kirtikar KR and Basu BD (1980). In: Indian MedicinalPlants.
Shirishadi 0.95 ± 0.95 ± 0.83 ± 0.76± 0.66 ± Singh B and Singh MP eds. Part II (p.937) India.
500mg/100g bwt 0.03** 0.03** 0.06** 0.06** 0.03** Misra LN, Dixit AK and Wagner H. N-demethyl
Endomethacin 0.72 ± 0.69 ± 0.67 ± 0.65 ± 0.66 ± budmunchiamines from Albizia lebbeck seeds. Phytochemistry. 1995;
0.03** 0.06** 0.06** 0.03** 0.03**
39(1): 247-249.
* Values are mean ± SEM (n =3 ); One-way ANOVA; df = 4, 10; 14,**P<0.001, Rashid RB, CHowdhury R, Jabbar A. Hasan CM and Rashid
*P<0.05 compared to control. MA. Constituents of Albizzia Lebbeck and antibacterial activity of an
isolated flavone derivatives. Saudi Pharmaceutical Journal. 2003; 11: 52-
STATISTICAL ANALYSIS
56.
 Journal of Applied Pharmaceutical Science 01 (10); 2011: 98-101

Ronaldo AR, Mariana LV, Sara MT, Adriana BPP, Steve P,

Ferreira SH and Fernando QC. Involvement of resident macrophages and
mast cells in the writhing nociceptive response induced by zymosan and
acetic acid in mice. Eur. J. Pharmacol. 2000;387: 111-118.
Shrotriya S, Ali MS, Saha A, Bachar SC and Islam MS. Anti-
inflammatory and analgesic effects of hedychium coronarium koen. Pak.
J. Pharm. Sci. 2007; 20(1): 42-47.
Vinegar R, Schreiber W and Hugo R. Biphasic development of
carrageenan edema in rats. J. Pharmacol. Exp. Ther. 1969;166: 96-103.
Winter CA, Risley EA and Nuss GW. Carrageenan induced Pharmacol. Chemotherp. 1964;22: 246-253.
edema in hind paw of the rat as an assay for anti-inflammatory drugs. World Health Organization. The global burden of disease: 2004
Proc. Soc. Exp. Biol. Med. 1962;111: 544-547. update. Geneva, WHO, 2008. Available at www.who.int/evidence/bod
Whittle BA. The use of changes in capillary permeability in World Health Organization. WHO factsheet 206: bronchial
mice to distinguish between narcotic and non-narcotic analgesics. Br. J. asthma. Available at: www.who.int/mediacentre/factsheets/fs206/en.