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Anti-inflammatory, analgesic and anti-pyretic activities of standardized root

extract of Jasminum sambac

Article  in  Journal of Ethnopharmacology · December 2014

DOI: 10.1016/j.jep.2014.11.039 · Source: PubMed

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 Journal of Ethnopharmacology 160 (2015) 140–148

Contents lists available at ScienceDirect

Journal of Ethnopharmacology
journal homepage: www.elsevier.com/locate/jep

Research Paper

Anti-inflammatory, analgesic and anti-pyretic activities of standardized

root extract of Jasminum sambac
Nidhi Sengar a, Apurva Joshi a, Satyendra K Prasad a,b, S Hemalatha a,n
a
Department of Pharmaceutics, Indian Institute of Technology (Banaras Hindu University), Varanasi, India
b
Department of Pharmaceutical Sciences, Rashtrasant Tukadoji Maharaj Nagpur University, Nagpur, Maharashtra, India

art ic l e i nf o a b s t r a c t

Article history: Ethnopharmacological relevance: The plant Jasminum sambac L. (Oleaceae) is cultivated throughout India.
Received 20 March 2014 The leaves and roots of the plant are used traditionally in the treatment of inflammation, fever and pain.
Received in revised form The leaves of the plant have been reported to posses significant anti-inflammatory and analgesic
13 October 2014
activities. Objective: To scientifically validate anti-inflammatory, analgesic and anti-pyretic activities of
Accepted 23 November 2014
roots from Jasminum sambac.
Available online 3 December 2014
Materials and methods: Ethanol root extract of Jasminum sambac (EJS) was standardized using HPTLC and
Keywords: was subjected to acute oral toxicity study. Further, analgesic activity of EJS at 100, 200 and 400 mg/kg,
Anti-inflammatory p.o. was evaluated using writhing test on Swiss albino mice and tail-flick test on Charles Foster albino
Analgesic
rats. Anti-inflammatory activity of EJS was assessed by carrageenan-induced rat paw edema, cotton
Anti-pyretic
pellet-induced granuloma and Freund's adjuvant-induced arthritis models, while antipyretic activity was
Jasminum sambac
Cotton pellet-induced granuloma evaluated using Brewer's yeast induced pyrexia. In addition, biochemical parameters such as alkaline
Freund's adjuvant phosphatase (ALP), aspartate transaminase (AST), alanine transaminase (ALT), lipid peroxidation (LPO),
superoxide dismutase (SOD) and catalase (CAT) in blood serum and edematous tissue of rats exposed to
acute (carrageenan) and granulomatous tissue in sub-chronic (cotton pellet granuloma) inflammation
models were also evaluated.
Results: Phytochemical analysis of EJS revealed the presence of flavonoids, phenols, saponins, tannins
and carbohydrates in major quantities, while the quantity of hesperidin in EJS (using HPTLC) was found
to be 4.25% w/w. EJS at 400 mg/kg, p.o. reduced writhing count up to 49.21%, whereas in tail-flick test, EJS
in a dose dependent manner increased latency in flicking tail. EJS at 400 mg/kg, p.o. showed significant
anti-inflammatory activity after 2nd, 3rd, 4th and 6th h of treatment in carrageenan-induced edema,
while a 33.58% inhibition in cotton pellet induced granuloma formation was observed at same dose level.
EJS significantly (po 0.001) inhibited adjuvant-induced arthritis and also showed significant antipyretic
activity. Further, a significant reversal in alterations of all the biochemical parameters (except ALP) in
tissues was also observed.
Conclusions: The study confirms the anti-inflammatory, analgesic and antipyretic activity of EJS which
may be attributed to the presence of various phytoconstituents quantified especially hesperidin which
have already been reported for its significant role in the treatment of inflammation and associated
problems.
& 2014 Elsevier Ireland Ltd. All rights reserved.

1. Introduction uptake, and the production of inflammatory mediators such as

As they are implied in virtually all human and animal diseases,
inflammation and pain have become the major focus of global
scientific research (Ibrahim et al., 2012). Inflammation is a com-
plex pathophysiological process mediated by a variety of signaling
molecules produced by leukocytes, macrophages and mast cells
undergoing various cellular responses. This includes phagocytic
n
Corresponding author. Tel.: þ 91 9415256481.
E-mail address: shemalatha.phe@itbhu.ac.in (S. Hemalatha).
nitric oxide (NO), prostaglandin (PGE2) and tumor necrosis factor
(TNF-α) (Kinne et al., 2000; Yu et al., 2010). These factors bring
about edema formation as a result of extravasations of fluids,
proteins and accumulation of leukocytes at the inflammatory site
(White, 1999). In addition, it is broadly accepted that cytokines,
produced by either immune or central nervous system cells, might
directly sensitize the peripheral nociceptors (Obreja et al., 2002).
Prostaglandins (PGs) induce hyperalgia by affecting the transdu-
cing property of free nerve endings, bradykinins, TNFα, interleu-
kins (ILs) and induce pain. Fever occurs due to infection produced
by generation of pyrogens including, ILs, TNF-α, interferon which

http://dx.doi.org/10.1016/j.jep.2014.11.039
0378-8741/& 2014 Elsevier Ireland Ltd. All rights reserved.
 N. Sengar et al. / Journal of Ethnopharmacology 160 (2015) 140–148
induces PGE2 production in hypothalamus and its temperature set
point. Pyrexia or fever is caused as a secondary impact of
inflammation (Khan et al., 2007). Inflammation, pain and fever
are all associated with enhanced production of prostaglandins
(Rang et al., 2001). Thus, most anti-inflammatory agents are also
expected to possess analgesic and antipyretic activities (Dewanjee
et al., 2009). Due to the adverse effects of non-steroidal anti-
inflammatory drugs (NSAIDs) and opioids, there is a high demand
for the search of new drugs with lesser or no side effects. In the
context, current trend of research has shifted towards medicinal
plants because of their affordability and accessibility with lesser
side effects (Ibrahim et al., 2012).
The plant Jasminum sambac L. (Oleaceae), commonly known as
Arabian jasmine is cultivated throughout India and tropical regions
of both hemispheres. The roots of the plant are used traditionally in
treatment of inflammation, fever and pain (Mishra, 2010; Abdoul-
Latif et al., 2010; AlRashdi et al., 2012). The flowers of the plant are
used for preparation of perfumes, aromatizing agents and in the
form of liana to decorate the hair. Traditionally, the plant is used as
analgesic, antidepressant, anti-inflammatory, antiseptic, aphrodisiac,
sedative, expectorant and uterine tonic. The essential oil from
Jasminum sambac is known as Jasmine oil, which is used as fragrance
for skin care products, in skin inflammation and as mood freshener
(Abdoul-Latif et al., 2010). Phytochemicals reported in this plant
includes dotriacontanoic acid, dotriacontanol, oleanolic acid, daucos-
terol, hesperidin. friedelin, quercitin, lupeol, betulin, α‐amyrin,
ursolic acid, sambacin, jasminin, sambacoside A, sambacolignoside,
isoquersitin, rutin, kaempferol, intecolin, linalool, α‐terpineol and
secoirridoid glucoside- sambacoside A‐G along with oleoside 11‐
methylester (AlRashdi et al., 2012; Sabharwal et al., 2011). Pharma-
cologically, the plant has been reported for antimicrobial (Al-
Hussaini and Mahasneh, 2011), puerperal lactation suppressant
(Shrivastav et al., 1988), antioxidant (Abdoul-Latif et al., 2010),
antidiabetic (Upaganlawer et al., 2009), hypolipidemic (Kalaiselvi
and Kalaivani, 2011), antiviral (Chiang et al., 2003), anti-acne (Tsai
et al., 2010), cardiac tonic (Somanadhan. et al., 1999), autonomic
nervous system stimulant (Naohiko et al., 2003) and antitumor
activities (Rahman et al., 2011). Also, the leaves of the plant have
been recently investigated for their significant anti-inflammatory
(Bhagat et al., 2007) and analgesic activities (Rahman et al., 2011).
Even though, the roots of the plant are traditionally used in
treatment of inflammation, fever and pain however, there is no
scientific data avalable on its traditional use. Therefore, on the basis
of the above evidences, present study was designed to scientifically
validate anti-inflammatory, analgesic and anti-pyretic activities of
roots from Jasminum sambac.
2. Material and methods
2.1. Plant material and extraction
The roots of the plant Jasminum sambac were collected from
herbal garden of Barchakha, South Campus, Banaras Hindu Uni-
versity, Varanasi, India and authenticated by Prof. N.K. Dubey
(Botanist), Department of Botany, Banaras Hindu University, Var-
anasi, India. The voucher specimen (No.: COG/JS/01) of the plant
has been deposited in Department of Pharmaceutics, Indian
Institute of Technology (Banaras Hindu University), Varanasi
(U.P), India for future reference. The roots of the plant were dried
in shade and powdered to obtain a coarse powder. The coarse
powder (500 g) was then macerated using ethanol (95% v/v) for a
week and the obtained extract was concentrated and evaporated
under reduced pressure in a rotary evaporator (IKA Germany) to
minimize the residual toxicity and were then kept in a desiccator
until use. The yield of the extract was found to be 13.589% w/w.
141
2.2. Chemicals and reagents
All the phytochemical markers such as diosgenin, D-fructose,
gallic acid, tannic acid, rutin, hesperidin and carrageenin were
purchased from Sigma-Aldrich, St. Louis, MO, USA and E. Merck,
Mumbai, India, while standard drug Indomethacin, diclofenac and
paracetamol were procured from IPCA Pharmaceuticals, Mumbai,
India. All the other chemicals and reagents were of analytical
grade and were purchased from Hi-Media Research Laboratories
Pvt. Ltd., Mumbai, India and S.D. fine Chemicals Pvt. Ltd., Mumbai,
India. All the spectrophotometric analysis was carried on Shi-
madzu, Pharmaspec-1700 ultraviolet-visible spectrophotometer.
2.3. Phytochemical standardization
Phytochemical screening of the ethanol root extract from
Jasminum sambac (EJS) was carried out for detection of various
phytoconstituents (Trease and Evans, 2002). Total carbohydrates in
plant extract were estimated by using anthrone reagent as
described by Yemm and Willis (1954). Total phenolic and tannin
content in extract were estimated according to Hagerman et al.
(2000) using Folin ciocalteau reagent, whereas total flavonoid and
flavonol contents were determined according to the methods of
Kumaran and Karunakaran (2006) using aluminum trichloride.
Total saponin content was estimated taking diosgenin as standard
by the implementing method of Baccou et al. (1977).
The presence of hesperidin in EJS was confirmed by thin layer
chromatography (TLC) using ethyl acetate:methanol:water:acetic
acid (25:2:2:1, v/v/v/v) as mobile phase. Further, EJS was standar-
dized for the first time with hesperidin using High Performance
Thin Layer Chromatography (HPTLC). A stock solution of extract
(5 mg/mL), and hesperidin (0.2 mg/mL) was prepared in methanol.
The mobile phase for developing the chromatogram was same as
used for TLC. The study was carried out using Camag-HPTLC
instrumentation (Camag, Mutten, Switzerland) equipped with
Linomat V sample applicator, Camag TLC scanner 3, Camag TLC
visualizer and WINCATS 4 software for data interpretation. The Rf
values were recorded and the developed plate was screened and
photo-documented at ultra violet range with wavelength (λmax) of
254 nm.
2.4. Experimental animals
Healthy Charles Foster albino rats (150–200 g) and Swiss albino
mice (20–30 g) of either sex were procured from the Central Animal
House (Reg. No. 542/02/ab/CPCSEA), Institute of Medical Sciences,
Banaras Hindu University, Varanasi, India. The certified pathogen
free animals were housed in polypropylene cages and maintained
under standard conditions (12 h light and dark cycle at an ambient
temperature of 25 71 1C and 45–55% RH). They were fed with
commercially available rat feed (Amrut Rat & Mice Feed Pvt. Ltd.,
Sangli, India) and water ad libitum. The animals were allowed to
acclimatize to the environment for 7 days before the commence-
ment of experiments. All experimental protocols were performed
after approval from Central Animal Ethical Committee of Banaras
Hindu University and were conducted in accordance with accepted
standard guidelines of National Institutes of Health Guide for Care
and Use of Laboratory Animals (Publication no. 85–23, revised 1985).
2.5. Acute toxicity study
Acute oral toxicity study was performed as per the guidelines of
Organization for Economic Co-operation and Development (OECD-
425). Nulliparous and non pregnant healthy female rats were used
for this study. The rats were divided into four groups with six
animals in each group. Single dose of EJS (500, 1000 and 2000 mg/kg
142 N. Sengar et al. / Journal of Ethnopharmacology 160 (2015) 140–148
p.o.) was administered to overnight fasted rats, while control group
revived distilled water (5 mL/kg). Animals were then observed
individually for 48 h for any behavioral and neurological changes
such as tremors, convulsions, salivation, diarrhea, sleep, lacrimation
and feeding behavior as a sign of acute toxicity for 48 h. The
observation was further extended up to 14 days to see any sign of
mortality. Further blood was collected from the rats and different
biochemical markers (total protein, total bilirubin, Alkaline phospha-
tase (ALP), Alanine transaminase (ALT), Aspartate transaminase (AST)
and urea) were estimated in blood serum. The animals were then
sacrificed by cervical dislocation and weights of different organs
were taken and compared with the control group (OECD, 2008).
2.6. Analgesic activity
2.6.1. Writhing test
For this study, mice (n ¼ 6) were divided into five groups, where
group 1 was served as vehicle control and was administered 0.5%
carboxy methyl cellulose (CMC). Group 2–4 were treated with EJS
at 100, 200 and 400 mg/kg, p.o., while group 5 was treated with
standard indomethacin at 10 mg/kg, p.o. After 30 min, writhing
was induced in mice by intraperitoneal injection of 0.6% acetic acid
(10 mL/kg, i.p.). The number of writhing was counted over a period
of 20 min, as previously reported (Amresha et al., 2007).
2.6.2. Tail-flick test
The method described by D’Amour and Smith (1941) was
adopted for this test, where grouping of animals was kept similar
to that of writhing test. The rat’s tail was placed at the window of
the tail flick apparatus (UGO Basile, Italy). Reaction times were
determined before and at 30, 60 and 90 min in both treated and
control groups. In this model, morphine (5 mg/kg, p.o.) was used
as standard drug.
2.7. Anti-inflammatory activity
2.7.1. Carrageenan-induced rat paw edema
For this study, standard drug used was diclofenac sodium at
10 mg/kg, p.o. After 30 min of extract administration, rats were
injected with 1% carrageenin into the plantar tissue of the right hind
paw. The contra-lateral hind paws were injected with 0.5% CMC as
control. Paw volume was measured plethysmographically at 0th, 1st,
2nd, 3rd, 4th and 6th h after carrageenan injection. After 6 h, rats
were sacrificed, the liver was dissected out and the blood was
obtained from all animals through retro-orbital plexus. The blood
samples were allowed to clot for 45 min at room temperature.
Serum was separated by centrifugation at 2500 rpm at 30 1C for
15 min and utilized for the estimation of various biochemical
parameters (Harris and Spencer, 1962).
2.7.2. Cotton pellet-induced granuloma
The granulomatous lesions was induced by surgically implant-
ing two cotton pellets subcutaneously in the dorsal region of the
rats, one near each axilla (grouping was same as in carrageenan-
induced rat paw edema). After 20 min of EJS administration,
autoclaved sterile pellets of cotton, weighing 20 71 mg each, were
aseptically implanted in the interscapular distance under the skin
on the previously shaved back of the rats in anesthetized condi-
tion. The rats were treated with extract and standard drug once
daily for 7 days and on the eighth day, all the rats were sacrificed
and the pellets surrounded by granuloma tissue were dissected
out carefully and dried at 70 1C. Mean weight of the granuloma
tissue formed around each pellet was recorded. The pellets were
weighed in both moist and dry condition. The weight of the pellets
taken out from EJS administered rats was compared with the
weight of pellets taken out from the control group (Swingle and
Shideman, 1972).
2.7.3. Freund's complete adjuvant-induced arthritis
Arthritis was induced to rats (among five groups as previously
divided) by a single intra-dermal injection of 0.1 mL of Freund's
complete adjuvant (FCA) containing 1.0 mg dry heat-killed Myco-
bacterium tuberculosis per milliliter sterile paraffin oil into a foot
pad of the left hind paw of male rats. A glass syringe (1 mL) with
the locking hubs and a 26 G needle was used for injection. The rats
were anesthetized with ether inhalation prior to and during
adjuvant injection, as the viscous nature of the adjuvant exerts
difficulty while injecting. The swelling in hind paws were periodi-
cally examined in each paw from the ankle using plethysmograph
on 0th, 7th, 14th, and 21st day after the treatment (Latha et al.,
1998).
2.8. Antipyretic activity
Rats of either sex were divided into five groups as before
containing six animals in each group for this experiment. The
normal body temperature of each rat was measured rectally at 1 h
interval on a thermometer. The antipyretic activity of extract was
evaluated using Brewer's yeast induced pyrexia in rats. Before
yeast injection, basal rectal temperature of rats was recorded and
was further given subcutaneous injection of 10 mL/kg of 15% w/v
yeast suspended in 0.5% w/v CMC solution for elevation of body
temperature of rats. At the 18 h after yeast injection, the vehicle,
standard drug and test drugs were administered, where paraceta-
mol 100 mg/kg, p.o. was taken as standard drug. Rectal tempera-
ture was recorded by digital thermometer at 0th, 1st, 2nd, 3rd and
4th h after drug administration (Asongalem et al., 2004).
2.9. Biochemical parameters
Blood serum and edematous tissue homogenate of rats (from
maximum effective dose of 400 mg/kg, p.o.), exposed to acute
(carrageenan) and granulomatous tissue in sub-chronic (cotton
pellet granuloma) inflammation models were centrifuged at
600  g for 10 min. The sediment, containing nuclei, unbroken
cells and plasma membranes (nuclear fraction) were separated
and the supernatant was subjected to centrifugation at 16,000  g
for 30 min. The sediment was suspended in 0.25 M sucrose buffer.
Aliquots were withdrawn at 0 and 30 min intervals, immediately
cooled at 0 1C and centrifuged at 16,000  g for 30 min. Enzyme
activity in the supernatant was then determined. Enzymes such as
alkaline phosphatase (ALP), aspartate transaminase (AST or SGOT),
alanine transaminase (ALT or SGPT) were assayed in the serum and
tissue homogenate by adopting standard methods (Reitman and
Frankel, 1957; Kind and King, 1954). In case of antioxidant studies
lipid peroxidation (LPO) was estimated by measuring thiobarbi-
turic acid reactive substances (TBARS) using the methods of
Nehius & Samuelson (1968). The activity of superoxide dismutase
(SOD) was assayed by the method of Kakkar et al. (1984), while
catalase (CAT) was estimated by the method of Sinha (1972).
2.10. Statistical analysis
The experimental results are expressed as mean7S.E.M., with six
animals in each group followed by one-way analysis of variance
(ANOVA). Newman–Keuls Multiple Comparision Test was applied for
determining the statistical significance between different groups.
However, two-way ANOVA followed by Bonferroni post test was
performed for determining the significance level in models such as
the tail flick method, carrageenan-induced rat padel-edema, adjuvant
 N. Sengar et al. / Journal of Ethnopharmacology 160 (2015) 140–148
induced arthritis and brewer's yeast induced hyperpyrexia. GraphPad
Prism (version 5) software was used for all statistical analysis.
P-value o0.05 was considered significant.
3. Results
3.1. Phytochemical standardization
Preliminary phytochemical analysis of EJS revealed the presence
of saponins, flavonoids, phenols, tannins, steroids and carbohy-
drates as major constituents. Total saponin quantified in the plant
material was found to be 126.82 mg/g diosgenin equivalent, while
total carbohydrate present in the plant extract was reported to be
108.422 mg/g D-fructose equivalent. Total phenolic content of EJS
was estimated to be 28.610 mg/g gallic acid equivalent, whereas
total tannin was reported to be 15.900 mg/g tannic acid equivalent.
Total flavonoid and flavonol content in the plant extract was found
to be 43.062 and 11.254 mg/g rutin equivalent. The HPTLC analysis
depicted well resolved peaks of EJS showing the presence of
hesperidin. The spots of the entire chromatogram were visualized
under UV 254 nm and the percentage of hesperidin (Rf 0.28) in EJS
was reported to be 4.25% w/w (Fig. 1).
3.2. Acute oral toxicity study
Acute oral toxicity study showed that, EJS up to 2000 mg/kg body
weight did not show any signs of behavioral or neurological toxicity.
A normal body weight gain was observed and there was no relative
difference in organ weights of control as well as treated rats. The
biochemical analysis also revealed no significant difference in the
levels of markers between the control and treated groups (Table 1).
3.3. Analgesic activity
As observed in Fig. 2, EJS at 200 and 400 mg/kg, p.o. and
diclofenac (10 mg/kg, p.o.) exhibited significant inhibition of the
writhes (po0.01, po0.001 and po0.001 respectively) when com-
pared to that of control. While, EJS at 100 mg/kg, p.o. did not show
significant analgesic effect. EJS at 400 mg/kg, p.o. reduced writhing
count up to 49.21% while diclofenac reduced the writhing count up
to 64.98%. Fig. 3 represents analgesic activity of EJS and morphine,
which did not showed any significant activity up to 30 min of
treatment. However, there was a significant effect observed, at all
dose level after 60 and 90 min of treatment showing an increased
latency in flicking tail. The observed effect was found to be more
pronounced (po0.001) in rats treated with EJS at 400 mg/kg, p.o.,
which was quite comparable with standard morphine.
Fig. 1. HPTLC chromatogram of hesperidin in ethanol extract of Jasminum sambac (EJS). (A) Standard peak of hesperidin. (B) Peak of hesperidin present in EJS.
143
3.4. Anti-inflammatory activity
The effect of EJS on carrageenan-induced rat paw edema is
shown in Table 2. The extract demonstrated a significant anti-
inflammatory effect against carrageenan-induced inflammation at
all doses level. EJS at 400 mg/kg, p.o. showed significant anti-
inflammatory activity after 2nd, 3rd, 4th and 6th h of treatment,
where maximum anti-inflammatory activity was observed at
4th h. The extract also illustrated a significant anti-inflammatory
effect against cotton pellet induced granuloma formation at 200
and 400 mg/kg, p.o. (p o0.05 and p o0.001 respectively). EJS at
100, 200 and 400 mg/kg, p.o. produced 3.7%, 5.93%, and 33.58%
inhibition of granuloma formation, whereas diclofenac produced
43.40% inhibition in granuloma formation (Fig. 4).
The effect of EJS on adjuvant induced arthritis is demonstrated
in Table 3. The adjuvant-induced arthritis was also significantly
inhibited as the paw volume reduced on treatment with EJS at
400 mg/kg, p.o. after 14th and 21st days of treatment (p o0.05).
The standard drug diclofenac (10 mg/kg, p.o.) showed significant
reduction in paw volume after 14th and 21st day (p o0.001 and
po 0.001 respectively).
3.5. Antipyretic activity
From the results (Table 4), it was observed that, experimental
rats showed a marked increase in rectal temperature, 18th h after
Brewer's Yeast injection. In the first hour EJS did not show anti-
pyretic activity. The antipyretic effect of EJS (200 mg/kg, p.o.) after
4th h showed significant (p o0.05) reduction in rectal tempera-
ture, while a significant reduction after 2nd, 3rd and 4th h
(p o0.05, p o0.01 and p o0.05 respectively) was reported with
EJS at 400 mg/kg, p.o. Antipyretic effect of paracetamol at 2nd, 3rd
and 4th h was significant (p o0.001) thus, it may be suggested
that EJS caused a highly significant reduction at third and fourth
hour of treatment, which is quite comparable with paracetamol.
3.6. Biochemical parameters
Table 5 represents results of different biochemical parameters
evaluated in the edematous and granulomatous tissues of acute and
sub-chronic anti-inflammatory studies. EJS depicted a significant
(po0.05) reduction in lipid peroxidation (LPO) in carrageenan-
induced rat pedal edematous tissue, whereas a highly significant
(po0.01) reduction in LPO level was observed in granulomatous
tissues of sub-chronic inflammation study. The extract showed a
significant increase in the levels of SOD and CAT in both models at
400 mg/kg, p.o. An inhibitory effect on AST and ALT was observed
with a significant value of po0.05 in both tested models. In
144 N. Sengar et al. / Journal of Ethnopharmacology 160 (2015) 140–148

Table 1
Effect of Jasminum sambac root extract (EJS) on various parameters evaluated in acute oral toxicity study.

Parameters Treatment groups

Control EJS 500 mg/kg EJS 1000 mg/kg EJS 2000 mg/kg

Body weight
0 day 175.250 7 3.293 183.2647 4.219 173.520 7 4.206 191.522 7 4.173
7th day 180.4417 4.057 188.552 7 4.736 183.7317 3.914 198.3347 5.795
14th day 186.7577 4.525 197.250 7 3.812 195.6737 5.266 211.1407 6.516
Organ weights
Heart 0.3497 0.011 0.360 7 0.010 0.3577 0.008 0.3557 0.012
Brain 0.450 7 0.047 0.4827 0.038 0.5177 0.042 0.4737 0.027
Liver 3.1687 0.321 3.2247 0.463 0.3177 0.242 0.326 7 0.226
Kidney (Right) 0.3127 0.017 0.308 7 0.011 0.3197 0.008 0.298 7 0.015
Kidney (Left) 0.294 7 0.021 0.286 7 0.015 0.2917 0.020 0.284 7 0.011
Lungs 0.4907 0.048 0.5717 0.032 0.580 7 0.027 0.522 7 0.043
Stomach 1.653 7 0.325 1.4847 0.216 1.5977 0.179 1.388 7 0.294
Biochemical analysis in blood serum
Total protein (g/L) 84.2617 1.781 78.2647 1.458 76.560 7 1.294 85.2797 2.041
Total bilirubin (mmol/L) 13.256 7 0.611 12.584 7 0.512 12.792 7 0.327 13.3417 0.773
ALP (U/mL) 67.265 7 1.279 63.1167 1.419 52.3527 1.219 62.7747 1.313
ALT (U/mL) 63.2577 1.086 58.3197 1.214 61.140 7 1.173 50.2047 1.357
AST (U/mL) 122.3647 1.356 110.5177 1.754 102.5677 1.285 97.086 7 1.552
Urea (mmol/L) 10.456 7 0.335 9.862 7 0.282 10.3177 0.516 9.0427 0.436

Values are mean7 SEM, (n¼ 6).

Fig. 2. Peripheral analgesic activity of Jasminum sambac root extract (EJS) in acetic
acid-induced writhing test. Values are mean7 SEM, (n ¼6), where ** corresponds
to po 0.01 and *** corresponds to p o 0.001 compared to control (In figure standard
corresponds to indomethacin at 10 mg/kg, p.o.).
contrast, there was no significant reduction observed in case of ALP
level in both tested models.
Results of different biochemical parameters evaluated in the
blood serum of acute and sub-chronic anti-inflammatory studies
are represented in Table 6. The results elicited a similar significant
(p o0.05) reduction in lipid peroxidation and increase in SOD and
CAT levels in serum of carrageenan-induced rat pedal edema and
cotton pellet induced granuloma model. From the observation, it
was found that a significant (p o0.05) decrease in ALT level was
reported only in case of serum of cotton pellet induced granuloma
model. AST and ALP levels estimated in both the models however,
showed inhibitory effect in their levels but were not found to be
significant. Standards used in both the models illustrated a sig-
nificant (po0.001) restoration of altered biochemical parameters.
4. Discussion
As inflammation, pain and fever are all attributed to elevated
levels of mainly prostaglandins, tumor necrosis factor and interleukins
(Rang et al., 2001; Khan et al., 2007), hence most anti-inflammatory
agents have also been shown to possess analgesic and antipyretic
Fig. 3. Central analgesic activity of Jasminum sambac root extract (EJS) in tail flick
method Values are mean 7 SEM, (n¼ 6), where * corresponds to p o0.05, **
corresponds to p o 0.01 and *** corresponds to p o 0.001 compared to control (In
figure standard corresponds to morphine at 5 mg/kg, p.o.).
activities (Dewanjee et al., 2009). The literature survey has revealed
that, the roots of the plant Jasminum sambac are being traditionally
used in treatment of inflammation, fever and pain (Abdoul-Latif et al.,
2010; AlRashdi et al., 2012). Therefore, the present investigation was
designed to scientifically validate the traditional claims of the roots
for the above proposed activities. In order to distinguish between the
central and peripheral analgesic action of Jasminum sambac, acetic
acid induced writhing response in mice was examined, which
depicted a dose dependent reduction in number of writhes. The
abdominal constriction is related to the sensitization of nociceptive
receptors to prostaglandins. It is therefore, possible that analgesic
effect of EJS may be due to inhibition in synthesis of prostaglandins
(Chen. 1993). The tail flick response appears to be a spinal reflex,
which is modulated by supraspinal inhibitory mechanism. EJS
demonstrated a significant (Po0.001) increase in latency to flick tail
at later phase in a dose dependent manner compared to control
group. The results suggested that, the analgesic effect of extract is
mediated through a peripheral mechanism by inhibition of prosta-
glandin synthesis (Nakamura et al., 1986).
Carrageenan is widely used as an inducing agent for experimental
inflammation for the screening of compounds possessing anti-
inflammatory activity. This phlogistic agent, when injected locally
into the rat paw, produces a severe inflammatory reaction, which is
discernible within 30 min (John and Nodine, 1999; Marzouk et al.,
 N. Sengar et al. / Journal of Ethnopharmacology 160 (2015) 140–148 145

Table 2
Anti-inflammatory activity of Jasminum sambac root extract (EJS) in carrageenan-induced rat paw edema.

Group Paw volume in mL (Mean 7 SEM)

0h 1h 2h 3h 4h 6h

Control 0.34 7 0.02 0.477 0.01 0.65 7 0.02 0.88 7 0.02 1.08 7 0.02 0.92 7 0.03
EJS 100 mg/kg 0.357 0.02 0.46 7 0.02 0.65 7 0.02 0.87 7 0.01 1.047 0.03 0.88 7 0.02
EJS 200 mg/kg 0.34 7 0.02 0.45 7 0.02 0.62 7 0.02 0.79 7 0.02n 0.977 0.04nn 0.84 7 0.02n
EJS 400 mg/kg 0.34 7 0.02 0.447 0.02 0.577 0.02n 0.617 0.01nnn 0.54 7 0.02nnn 0.477 0.02nnn
Diclofenac (10 mg/kg, p.o.) 0.337 0.02 0.40 7 0.02 0.45 7 0.02nnn 0.52 7 0.02nnn 0.46 7 0.02nnn 0.417 0.01nnn

Values are mean7 SEM, (n¼ 6), where

n
corresponds to po 0.05,
nn
corresponds to p o0.01 and
nnn
corresponds to p o0.001 compared to control.

chronic inflammatory reaction and can serve as a sub-chronic

inflammatory test model. This model has been employed to assess
the transudative and proliferative components of chronic inflamma-
tion. The fluid adsorbed by the pellet greatly influences the wet
weight of the granuloma, whereas the dry weight correlates well
with the amount of granulomatous tissue formed (Panthong
et al., 2003). The extract showed decrease in granuloma formation
that reflected its efficacy to reduce increased level of fibroblasts and
synthesis of collagen with mucopolysaccharide, which are natural
proliferative events of granulation tissue formation. (Panthong et al.,
2003). Literatures have revealed that complete Freund's adjuvant
induces arthritis, where both T cell and B cell activation is important
in collagen induced arthritis. Cytokines of both Th1 (IFNγ secreting)
and Th2 cells (IL-4 secreting) are produced, and at disease onset a
Fig. 4. Anti-inflammatory activity of Jasminum sambac root extract (EJS) on cotton Th1 profile predominates (Courtenay et al., 1980). Thus, the observed
pellet induced granuloma formation. Values are mean 7SEM, (n ¼6), where *
anti- inflammatory activity of EJS may also be attributed to inhibitory
corresponds to p o 0.05 and *** corresponds to p o0.001 compared to control (In
figure standard corresponds to Diclofenac at 10 mg/kg, p.o.). effects on cytokines.
The subcutaneous injection of yeast suspension markedly ele-
vated the rectal temperature after 18 h of its administration however;
Table 3 on treatment with EJS a significant reduction in the rectal tempera-
Anti-inflammatory activity of Jasminum sambac root extract (EJS) in adjuvant ture was observed. At lower dose, EJS (100 mg/kg, p.o.) was found to
induced arthritis.
be non significant, while EJS at higher dose showed prominent
Group Paw volume in mL (Mean 7 SEM) activity and reduced the rectal temperature at 2nd, 3rd and 4th h
(po0.05, po0.01,and po0.01 respectively). This results confirmed
0h 14 day 21 day that, Jasminum sambac extract have an influence on prostaglandin
biosynthesis, since prostaglandin is believed to be a regulator of body
Control 0.3417 0.002 0.7377 0.001 0.753 7 0.003
EJS 100 mg/kg 0.3377 0.001 0.7347 0.003 0.750 7 0.003 temperature (Dascombe, 1985).
EJS 200 mg/kg 0.3397 0.002 0.7287 0.002 0.7437 0.001 Marker enzymes and antioxidant activity were assayed in
EJS 400 mg/kg 0.3357 0.004 0.7267 0.003n 0.7407 0.002n serum and different target tissues, of rats treated with EJS to
Diclofenac (10 mg/kg, p.o.) 0.3387 0.005 0.7237 0.002nn 0.540 7 0.006nnn investigate the involvement of antioxidant mechanism in the
Values are mean7 SEM, (n¼ 6), where
protective effect of Jasminum sambac in inflammation. Formations
n
corresponds to po 0.05,
of malondialdehyde (MDA), marker index for lipid peroxidation,
nn
corresponds to p o0.01 and were significantly increased in edematous, granulomatous tissues
nnn
corresponds to p o0.001 compared to control. and also in serum during hind paw edema and cotton pellet
granuloma. Maximum effective dose of EJS (400 mg/kg, p.o.)
2010). Carrageenan-induced hind paw edema in rat is a biphasic significantly reduced the elevated MDA levels in different target
event. The early phase (90–180 min) of the inflammation is due to tissues and also in serum. In such types of proliferative process,
the release of histamine, serotonin and similar substances; and the excessive formation of free radicals occurs, which triggers membrane
later phase (270–360 min) is associated with the activation of kinin- damage and also augments activation of membrane bound enzymes
like substances, i.e. prostaglandins, proteases and lysosome (Yadav et al., 2005). As observed, there was a marked decline in SOD
(Thomazzi et al., 2010). The lower dose i.e. EJS 100 mg/kg, p.o. and CAT levels of edematous, granulomatous tissues and serum of
showed inhibition at late phase (po0.05), while the higher dose control group rats in hind paw edema and cotton pellet granuloma
400 mg/kg, p.o. showed significant activity at early as well as late models however, treatment with EJS, significantly restored the
phase. The standard drug diclofenac showed maximum activity at depleted levels of SOD and CAT.
early phase (Po0.01). The carrageenan-induced paw edema test is AST, ALT and ALP enzymes were significantly elevated in both
effectively controlled with the arachidonate cyclooxygenase (COX) acute and sub-chronic inflammation. In edematous and granuloma-
inhibitors due to its COX-dependent mechanism, thus, from the tous tissue, EJS at 400 mg/kg, p.o. significantly restored the AST and
observed results, it is suggested that EJS may possess arachidonate ALT level but did not alter ALP level. In blood serum, EJS did not alter
COX inhibitory property (Panthong et al., 2003). Cotton pellet- any of the serum markers in paw edema, while in cotton pellet, EJS
induced granuloma formation is a typical feature of an established only elevated ALT level. Apparently, it appears that the membrane
146 N. Sengar et al. / Journal of Ethnopharmacology 160 (2015) 140–148

Table 4
Effect of Jasminum sambac root extract (EJS) on Brewer's yeast induced hyperpyrexia.

Group Temperature in 1C

Initial. 0h 1h 2h 3h 4h

Control 36.337 0.21 40.177 0.48 40.177 0.48 40.007 0.26 39.83 70.31 39.6770.21
EJS 100 mg/kg 36.50 7 0.22 40.007 0.26 39.83 7 0.31 39.83 7 0.48 39.6770.21 39.50 70.62
EJS 200 mg/kg 36.007 0.26 40.337 0.56 39.677 0.21 39.677 0.43 39.50 70.43 38.17 70.17n
EJS 400 mg/kg 36.177 0.65 39.677 0.33 39.177 0.31 38.337 0.21n 38.00 70.26nn 37.6770.21nn
Paracetamol (100 mg/kg, p.o.) 36.177 0.40 40.177 0.31 39.50 7 0.22 37.007 0.26nnn 36.83 70.48nnn 36.3370.33nnn

Values are mean7 SEM, (n¼ 6), where

n
corresponds to po 0.05,
nn
corresponds to po 0.01 and
nnn
corresponds to p o0.001 compared to control.

Table 5
Effect of Jasminum sambac root extract (EJS) on various biochemical parameters in dry edematous tissue of rats exposed to acute (carrageenan) and granulomatous tissue in
sub-chronic (cotton pellet granuloma) inflammation models.

Acute inflammation (carrageenan) model Sub-chronic inflammation (cotton pellet granuloma)

model

Biochemical parameters Control EJS 400 mg/kg Diclofenac (10 mg/kg, p.o.) Control EJS 400 mg/kg Diclofenac (10 mg/kg, p.o.)
AST (Units/mL) 86.50 71.38 81.337 0.88n 59.337 1.31nnn 91.177 1.25 83.83 71.89n 64.837 1.62nnn
ALT (Units/mL) 84.17 71.14 79.677 1.15n 60.177 0.65 nnn
96.50 7 1.26 91.50 71.69n 64.837 1.17nnn
ALP (Units/mL) 69.00 71.24 66.177 1.54 60.50 7 1.36nn 70.177 1.45 63.83 71.76 56.337 1.31nn
SOD (Units/mg of protein) 7.3370.49 9.177 0.48n 11.337 0.49nnn 7.83 7 0.48 10.6770.49nn 12.177 0.75nnn
CAT (mmol H2O2 consumed/min/mg of protein) 64.61 73.92 87.517 2.75nnn 98.89 7 2.82nnn 67.667 4.39 95.23 73.94nnn 103.87 7 3.86nnn
TBARS (Units in mole/mg of protein) 100.3371.38 94.177 1.87n 74.177 1.14nnn 92.83 7 1.76 85.6771.20nn 66.177 1.42nnn

Values are mean 7SEM, (n ¼6), where

n
corresponds to p o0.05,
nn
corresponds to p o0.01 and
nnn
corresponds to p o 0.001 compared to control.

Table 6
Effect of Jasminum sambac root extract (EJS) on various biochemical parameters in serum of rats exposed to acute (carrageenan) and sub-chronic (cotton pellet granuloma)
inflammation models.

Acute inflammation (carrageenan) model Sub-chronic inflammation (cotton pellet granuloma)

model

Biochemical parameters Control EJS 400 mg/kg Diclofenac (10 mg/kg, p.o.) Control EJS 400 mg/kg Diclofenac (10 mg/kg, p.o.)
AST (Units/mL) 100.007 1.29 102.17 71.62 89.677 2.87nn 107.50 7 1.61 102.17 71.92 64.507 1.12nnn
nn
ALT (Units/mL) 80.83 7 1.58 77.3370.76 72.677 2.09 91.677 1.48 85.6771.80n 73.83 7 1.70nnn
ALP (Units/mL) 86.337 1.65 83.83 71.54 60.677 1.02nnn 91.337 1.54 88.50 71.09 60.677 1.02nnn
SOD (Units/mg of protein) 6.83 7 0.60 9.17 70.48n 12.337 0.76nnn 7.83 7 0.31 9.6770.33n 12.677 0.76nnn
CAT (mmol H2O2 consumed/min/mg of protein) 56.87 7 2.88 68.21 72.36nn 86.157 2.13nnn 60.75 7 3.71 75.30 72.01nn 93.94 7 2.34nnn
TBARS (Units in mole/mg of protein) 100.337 1.38 94.3371.36n 88.83 7 1.51nnn 100.677 1.31 95.50 71.73n 75.177 1.22nnn

Values are mean 7SEM, (n ¼6), where

n
corresponds to p o0.05,
nn
corresponds to p o0.01 and
nnn
corresponds to p o 0.001 compared to control.

damage seems to be the prime culprit for the marked increase in the
serum marker enzymes AST, ALT and ALP. There are increasing
evidences that, lysosomal enzymes play an important role in the
development of acute and chronic inflammation (Anderson et al.,
1971). Most of the anti-inflammatory drugs exert their beneficial
effects by inhibiting either release of these enzymes or by stabilizing
lysosomal membrane, which is one of the major events responsible
for the inflammatory process (Nair et al., 1988). Thus, it may be
presumed that, EJS extract may have an inhibitory effect on liposomal
enzymes or may act by stabilizing the membrane.
Preliminary phytochemical analysis and quantitative evaluations of
EJS demonstrated the presence of mainly phenols, flavonoids, sapo-
nins, tannins and carbohydrates. Available literature has revealed the
role of these phytochemicals in treatment of inflammation and
associated problems (Liu and Lin. 2012; Piwowarski et al., 2011;
Yang et al., 2013; Nalbantsoy et al., 2012). Hesperidin, as previously
reported in the roots of Jasminum sambac (Abdoul-Latif et al., 2010) is a
naturally occurring flavonoid which exerts wide range of pharmaco-
logical activities such as antioxidant, anti-inflammatory, antihyperch-
olesterolemic and anticarcinogenic actions with different mechanism
(Ahmadi et al., 2008). A recent study has confirmed the potential role
of hesperidin in reversing inflammatory conditions as observed
through TNF-a, total leukocytes, neutrophils lymphocytes and nitrite
estimations in a rat air pouch model of inflammation. It has also been
shown to reverse alterations in the levels of LPO, GSH, SOD, and CAT in
inflammation models (Jain and Parmar, 2011; Ahmadi et al., 2008).
Hesperidin has also been reported for its significant role in treatment
of arthritis (Guardia et al., 2001). Thus, the presence of hesperidin in
EJS may act as a major contributing factor in its observed anti-
inflammatory, anti-nociceptive and antipyretic activities.
 N. Sengar et al. / Journal of Ethnopharmacology 160 (2015) 140–148
5. Conclusion
In conclusion, Jasminum sambac root extract exhibited signifi-
cant anti-inflammatory activity at both early and late phase. The
activity may be due to a cumulative effect of hesperidin and other
phytoconstituents. The study also confirms its anti-nociceptive
and antipyretic activities. Thus, the present study scientifically
validated the traditional use of roots from Jasminum sambac in
treatment of pain and inflammatory related ailments.
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