[Pharmaceutical Biology (2002): (40), 6, 416-421]
Antispasmodic Activity of SJ-200 (Himcospaz) An Herbal Preparation
M.V. Venkataranganna, S.D. Anturlikar, S. Gopumadhavan, N.S. Prakash,
Mohammed Rafiq, Girish S. Murthy and S.K. Mitra*
R&D Center, The Himalaya Drug Company, Makali, Bangalore 562 123 (India).
SJ-200 (Himcospaz), an herbal preparation, was investigated for antispasmodic activity on
gastrointestinal smooth muscles of guinea pig, rats, rabbits, and mice. SJ-200 dose-
dependently inhibited acetylcholine, histamine and barium chloride-induced contraction of
guinea pig ileum. It inhibited spontaneous contraction of rabbit and rat colon. Oxytocin-
induced contraction of rat uterus was also inhibited. Oral administration of SJ-200 dose-
dependently reduced gastric emptying in rats and intestinal transit in mice. All these findings
suggest the non-specific antispasmodic activity of SJ-200 in experimental models.
Keywords: Antispasmodic, gastric emptying, intestinal transit, irritable bowel syndrome,
Functional gastrointestinal disorders (FGID) affect millions of people from all age groups
(Loe et al., 1999). Irritable bowel syndrome (IBS), dyspepsia and inflammatory bowel
disease (IBD) are the most common disorders of FGID. The symptoms of FGID can cause
discomfort, ranging from inconvenience to deep personal distress (Koloski et al., 2000).
Unfortunately no single drug has proven to be effective in treating FGID. In addition, the
search for a truly effective and safe drug to control motility disturbances in FGID continues
(Scapignato & Pelosini, 1999).
Ayurveda, an Indian system of medicine, cited several plants, which are useful against
various gastrointestinal disorders without any side effects. SJ-200 (Himcospaz), an herbal
preparation contains Zingiber officinale Roscoe, Zingiberaceae (rhizome), Apium graveolens
L., Apiaceae (fruit) and Foeniculum vulgare Mill., Apiaceae (fruit). All these plants have
been used to treat various gastrointestinal disorders like abdominal pain, flatulence and colic
(Satyavathi, 1976; Nadkarni, 1982; Vavier,1996).
In the present study we have investigated the effect of SJ-200 on various smooth muscles in
vitro, gastric emptying rate and intestinal transit rate in vivo.
MATERIALS AND METHODS
Apium graveolens, Foeniculum vulgare and Zingiber officinale were procured from a local
supplier and identified by Dr. Kannan, Botanist, The Himalaya Drug Company, Bangalore.
Samples were retained for reference purpose at the R & D herbarium.
The drugs acetylcholine chloride (Ach) and histamine dihydrochloride were from Sigma
Chemical Co., St. Louis. MO. Oxytocin was from Rathi Laboratories (Hindustan Pvt. Ltd.,
Patna, India). All other chemicals were purchased from Loba Chemie, Mumbai, India.
Preparation of physiological solutions
Tyrode solution composition (mM): NaCl, 137; KCl, 2.7; CaCl2, 1.8; NaH2PO4, 0.4; MgSO4
0.25; NaHCO3, 11.9; glucose, 11.1 de Jalon solution composition (mM): NaCl, 154; KCl,
5.6; CaCl2, 0.55; NaHCO3, 6.0; glucose 2.78.
Preparation and standardization of SJ-200
Dried and powdered materials of Zingiber officinale, Apium graveolens and Foeniculum
vulgare in the ratio of 1:1:1 were mixed and 100 g powder was refluxed with 500 ml
chloroform at 70°C for 6 h. After filtration the chloroform was evaporated in a vacuum
evaporator. The extract thus obtained was taken for the standardization and pharmacological
studies. The yield of the extract was 10% of total dried material.
Ten microlitres of 1% v/v solution of SJ-200 in
chloroform was injected into a 20 M stainless-
steel column filled with 10% carbowax. Nitrogen
was used as the carrier gas at 30 ml per min and
a flame ionization detector was used. The
temperature of the oven, injector and detector
were 220, 230 and 250oC respectively. The
chromatogram was recorded with the help of
Aimil chromatography data station. A fingerprint
chromatogram of SJ-200 is presented in Fig. 1.
igure 1. Gas chromatogram of SJ-200
Swiss albino mice, albino rats (Wistar strain),
New Zealand white rabbits and guinea pigs were
housed in an air-conditioned area at 25 ± 2oC
with 12:12 h light and dark cycle. They were
maintained on synthetic pelleted feed (Lipton
India Ltd., Mumbai, India) and water ad libitum.
Animals for organ bath studies
Male and female Wistar rats (160-200 g b.wt.) and male guinea pig (300-350 g b.wt.) after 24
h fasting were killed by stunning and bleeding. Ileum, colon or uterus were removed and
suspended under a constant tension of 1 g in 15 ml organ baths containing Tyrode at 37°C
(Ulrike et al., 1997; Parry et al.,1996). Before the experimental procedures the organs were
allowed a 30-40 min equilibration period with changes of medium at every 5 min. SJ-200 was
dissolved in 0.5% DMSO. 200 µl diluted SJ-200 was used as the maximum volume in the
baths, at which volume there was no solvent effect. During the experimental phase SJ-200
was added to the bath and after 1 min incubation period the agonist was added. Contractions
were recorded using isotonic transducer connected to a polyrite (Model 201, Recorders and
Isolated rat colon and uterus
A portion (2.5 cm) of colon (n=6) was washed and suspended in aerated tyrode solution at
37°C. The uterus (n=6) from an estradiol-treated female rat (10 µg/kg s.c. 48 h before
experiment) was isolated and suspended in de Jalon solution (Blazquez et al.,1995).
Isolated guinea pig ileum and rabbit colon
A portion of 2.5 cm guinea pig ileum (n=6) and rabbit colon (n=3) was suspended in aerated
Tyrode solution at 37°C.
Effect of drugs
Dose response curves or isolated concentrations were performed with Ach (10-8-10-4 M),
Oxytocin (2 mIU/ml) histamine 10-8-10-4 M and barium chloride (10-3–10-1 M) in the
presence or absence of SJ-200. The effect of SJ-200 on normal motility of rat and rabbit
colon was evaluated without agonists.
Effect of SJ-200 on gastric transit rate in rats
Wistar strain rats of either sex weighing between 180-200 b.wt. were used for the study.
Twenty -four overnight fasted rats were divided into 4 groups of 6 each. Group I served as
control and received vehicle alone. Group II, III and IV received SJ-200 at a dose of 50, 100
and 200 mg/kg p.o. in 0.5% DMSO. After 30 min all the rats received 1.5 ml of test
meal/animal, which contains 50 mg of phenol red in 100 ml of 1.5% w/v aqueous
methylcellulose. Twenty minutes after administration of the test meal, the rats were
euthanised under ether anesthesia. The stomach was dissected out by opening the abdominal
cavity after clamping the cardiac and pyloric ends. The stomach was rinsed in physiological
saline and placed into 100 ml 0.1 NaOH and cut into small pieces and then homogenized. The
homogenized suspension was allowed to settle for 60 min at room temperature. Five milliliter
of supernatant was pipetted out into a test tube to which 0.5 ml of 20% w/v
trichloroaceticacid (TCA) was then added and centrifuged at 2800 rpm for 20 min. To the
supernatant 4 ml of 0.5N NaOH was added and absorbance was measured
spectrophotometrically at 560 nm. For the standard concentration, phenol red recovered from
the stomach of rats sacrificed immediately following oral administration was considered
(Tache et al., 1987).
Effect of SJ-200 in intestinal transit time in mice
Twenty-four Swiss albino mice weighing 25-28 g were fasted overnight and divided into four
groups of 6 each. Group I served as control, which received vehicle alone. Group II, III and
IV received SJ-200 at a dose of 50, 100 and 200 mg/kg p.o. in 0.5% DMSO. After 30 min all
the mice received charcoal meal (1.0% charcoal in 1.5% tragacanth) at a dose of 0.1
ml/animal. Twenty minutes after administration of the charcoal meal, the intestine was
removed. The total length of the intestine, from duodenal end to the cecum, and the distance
travelled by the charcoal were recorded and the percentage of charcoal movement was
calculated (Hamada et al., 1999).
Data were expressed as Mean ± SEM. Significance was assessed by Student’s ‘t’ test or
ANOVA followed by Dunnet’s test. The minimum level of significance was fixed at p<0.05.
Effect of SJ-200 on spontaneous motility of rat and rabbit colon
SJ-200 at a concentration of 40-80 µg/ml dose-dependently inhibited spontaneous contraction
of rat and rabbit colon. A dose of 80 µg/ml of bath concentration completely blocked
spontaneous contraction (Figs. 2 and 3).
igure 2. Effect of SJ-200 on normal motility of rat colon.
igure 3. Effect of SJ-200 on normal motility of rabbit colon.
Effect of SJ-200 on agonist-induced
Acetylcholine and histamine (10-8-10-4
M) induced dose-dependent contraction
of guinea pig ileum. Exposure (60 sec)
of the ileum to SJ-200 (40-80 µg/ml)
reduced the contractile response of
acetylcholine and histamine (10-4 M) in
a dose-dependent manner (Fig. 4).
SJ-200 at a dose of 80 µg/ml almost
completely blocked the barium
chloride-induced contraction of guinea
pig ileum (Fig. 5). The incubation of
igure 4. Effect of SJ-200 on acetylcholine- and histamine-
induced contractions of guinea pig ileum
the isolated rat uterus with SJ-200 (80 µg/ml) resulted in significant inhibition of the
oxytocin-induced contractile response (Fig. 6).
igure 5. Effect of SJ-200 on barium chloride (BaCl2)-induced contractions of guinea pig ileum.
igure 6. Effect of SJ-200 on oxytocin (OXT)-induced contractions of rat uterus.
Effect of SJ-200 on gastric
emptying rate (GER) and intestinal
transit time (ITT)
SJ-200 (50-200 mg/kg b.wt) dose-
dependently reduced gastric emptying
and intestinal transit in rat and mice
respectively. The effects on both
parameters were significant at a dose
of 100 and 200 mg/kg b. wt. p.o. of
SJ-200 (Fig. 7).
igure 7. Effect of SJ-200 on gastric emptying in rats
The present study shows that SJ-200
reduces spontaneous motility of rat
and rabbit colon. The absence of
contractile activity of the drug itself
on various smooth muscle
preparations shows the lack of
agonistic activity on muscaranic,
histaminergic and oxytocin receptors.
The results obtained on isolated
smooth muscle show the inhibition of
histamine, acetylcholine, oxytocin
and barium chloride-induced contractions.
igure 8. Effect of SJ-200 on intestinal transit in mice
Binding of Ach to muscarinic receptors or histamine to H1 receptor in smooth muscles results
in opening of receptor operated channels, thereby allowing sodium influx, which causes a
depolarization of the cell membrane. This depolarization opens voltage dependent calcium
channels and calcium ions enter the cell to induce the release of calcium from the
sarcoplasmic reticulum. The cytosolic calcium thus binds to calmodulin, which results in
contraction (Balton, 1979; Rojas et al., 1996).
Since these spasmogens have different modes of action, the antagonism elicited by SJ-200
indicates that it might be acting at a common step in the contraction mechanism elicited by
these agonists. The antagonism displayed was concentration dependent. Since acetylcholine,
histamine, oxytocin and barium chloride effects were altered by the SJ-200, it seems to be
non-specific antagonism. Earlier research works on the extracts of individual ingredients of
Zingiber officinale, Apium graveolens and Foeniculum vulgare were credited for their
antispasmodic activity. Zingiber officinale is proven to be effective in inhibiting the gastric
and intestinal motility in mice and also found to inhibit the colonic motility in rats (Tulimat et
al., 2001; Wu et al., 1994). The spasmolytic activity of Zingiber officinale could be attributed
to gingerol, an active constituent that was found to inhibit prostaglandin biosynthesis and
seratonergic activity (Harborne et al., 1999). Apigenin, an active constituent of Apium
graveolens, was reported to inhibit norepinephrine-induced contractions of rat aortic
preparations in a dose-dependent manner (Ko et al., 1991). The essential oils of Foeniculum
vulgare are reported to exhibit antispasmodic effect in rat uterus preparation. (Ostad et al.,
2001; Neuhas-Carlisle et al., 1993; Forster et al., 1980). The combination of these active
constituents could be responsible for the observed antispasmodic effect of SJ-200.
Irritable bowel syndrome is the most common functional disorder, which is characterized by
a combination of abdominal pain and altered bowel function affecting primarily the mid and
lower gut (Vasllo et al., 1992). As a consequence drugs affecting gastrointestinal motility
have been widely employed with the aim of correcting the major IBS manifestation i.e. pain
and altered bowel function (Scapignato & Pelosini, 1999). The present study reveals that SJ-
200 dose-dependently decreased gastric emptying and intestinal transit, which indicates
inhibition of gastro-intestinal motility in vivo. The in vivo observations were also correlating
with in vitro studies. All these findings suggest that SJ-200 is having non-specific
antispasmodic activity, which can be used in the treatment of various non-specific spasm
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