Effect Of Siddha Drug (Kantha Chendooram) OnDocument Transcript
J. Res. Educ. Indian Med., July - Sept., 2008
EFFECT OF SIDDHA DRUG (KANTHA CHENDOORAM) ON
INDOMETHACIN INDUCED GASTRIC ULCER LESIONS IN RATS
R.VADIVELAN,1 K.ELANGO,1 B.SURESH,1 B.R.RAMESH,1
S.BHADRA,1 A.RAGHURAM1 AND R.SAMPATHKUMAR2
Department of Pharmacology,1 J.S.S. College of Pharmacy, Ooty - 643001 Tamilnadu (India)
Govt. Siddha Medical College,2 Palayamkottai, Tirunelveli - 627002 Tamilnadu (India)
Abstract : To assess the possible anti-ulcer effect of Kantha Chendooram,ulcer index,
ulcer protection, non-protein sulphydryls (NP-SH) and adherent mucus content were
determined in indomethacin induced gastric mucosal injury in rats. Pretreatment with
Kantha Chendooram significantly prevented the gastric mucosal lesion development and
decreased the gastric toxicity produced by ulcerogen. In addition, ulcerated rats showed
depletion of gastric wall mucus and NP-SH levels whereas treatment with Kantha
Chendooram reverted this decline in indomethacin induced rats. Histological studies
confirmed the results. The present finding suggests that Kantha Chendooram promotes
ulcer protection by decrease in ulcer index and increase in gastric mucin content and NP-
SH concentration. Kantha Chendooram may protect the gastric mucosa against ulceration
by its cytoprotective activity.
Keywords : Antiulcer activity, Kantha Chendooram, Indomethacin, Histopathology,
Mucus content, Siddha drug.
Introduction Indian Medical Practitioners Co-operative
Kantha Chendooram is a popular Siddha Pharmacy and Stores Ltd., (IMCOPS)
preparation of eight ingredients indicated for Tirunelveli, Tamilnadu, India.
microcytic anemia, anemia, chlorosis, obesity,
edema, scrotal swellings, and rheumatic Haemoglobin, alcian blue, tricholoroacetic
diseases, enlargement of liver and spleen and acid, sodium acetate, sucrase, 5, 5 -dithio-bis
abdominal tumors. It consists of Purified Lode dinitro-benzoic acid (DTNB) were all purchased
Stone (Suththi seitha kantham), Purified Sulphur from the Sigma Chemical Company. All other
(Suththi seitha kanthakam), Lead Wort root reagents used for the experiment were of
powder (Koduveliver podi), Eclipta juice analytical grade.
(Karisalaisaru), Lime juice (Elumicham pazha
saru), Milk (Paal), Egg albumin (Muttaiyin Animals
venkaru), Mudar Latex (Erukkan paal) Healthy adult albino rats of Wistar strain
(Formulary of Siddha Medicines, 1993). weighing 180-250g were obtained from J.S.S.
In the present study an attempt has been made College of Pharmacy, Animal House, Ooty,
to validate the anti-ulcer activity of standardized India. The animal house was well ventilated
Kantha Chendooram. and animals had 12±1 hour day and night
schedule with temperature between 11-20±2oC.
Materials and Methods The animals were housed in large spacious
Drugs and Chemicals hygienic cages during the course of the
Kantha Chendooram was procured from experimental period. The animals were fed with
24 Vadivelan et al.
rat pellet feed supplied by M/s. Hindustan Lever Ulcer Descriptive
Ltd., Bangalore, India and water ad libitum. Score Observation
Six animals were used in each group. 0 Normal
1 Less than 1mm (Pin point)
The protocol was approved by Institutional 2 1-2 mm
Animal Ethics Committee constituted for the 3 Greater than 2 mm and above
purpose. We have converted the human dose
of Siddha drug into animal dose as per the
standard surface ratio method. (Ghosh M.N., The ulcer score was divided by a factor
1984) of 10 to get the ulcer index. % Ulcer protection
was calculated according to the standard
Indomethacin-induced gastric mucosal formula. (Njar et al., 1994)
injury in rats (Suleyman et al., 2001)
Adult Wistar albino rats of either sex Ulcer Index in Control – Ulcer index in Test
weighing 180-250 g were divided into four groups —————————————— X 100
of six animals each and placed in cages with Ulcer Index in Control
grating floor to avoid coprophagy and fasted
for 24 hours allowing free access of water. Gastric Mucosal Defensive Factors
Estimation of Mucous barrier (Kulkarni et
Group I served as Solvent Control al., 1996)
(0.3% Carboxy Methyl Glandular portions of stomach of 24 hrs
Cellulose Sodium) fasted rats were everted and soaked for 24 hrs
Group II received Sucralfate in 10 ml of 0.1% alcian blue 8GX dissolved in
(270 mg/kg) 0.16 M sucrose buffered with 0.05 M sodium
Group III received Kantha Chendooram acetate adjusted to pH 5.8 with Hcl.
(20 mg/kg) Uncomplexed dye was removed by two
Group IV received Kantha Chendooram successive washes of 15 and 45 minutes in
(40 mg/kg) 0.25 N sucrose. Dye complexed with mucous
was dilated by immersion in 10 ml aliquots of
The standard drugs and test drugs were 0.5 M magnesium chloride for 2 hrs.
administered orally half an hour before the oral The resulting blue solutions were shaken with
administration of indomethacin (25 mg/kg) to equal volume of diethyl ether and optical density
the 24 hours fasted rats. Six hours later the of aqueous phase was measured at 605 nm.
animals were sacrificed using excess ether The barrier mucous was expressed in terms of
anaesthesia. The stomach was excised carefully, microgram of alcian blue dye/g of wet stomach
opened along the greater curvature; the luminal glandular tissues.
contents were removed. The mucosa was
flushed with saline and the stomach pinned on Mucous barrier [(microgram of alcian
a frog board. blue dye/g of wet stomach glandular tissues
The ulcer index was calculated according to Absorbance x 105
the method. (Asuzu and Omu, 1990) The lesions = ———————————————
were counted with the aid of hand lens (10X) and E1% 1cm x wt. of glandular tissues
each given a severity rating as follows: E1% for alcian blue = 189
Effect of Kantha Chendooram in Gastric Ulcer 25
Estimation of Non–Protein Sulf hydryl (NP- by one way Analysis of Variance (one way
SH) Group (Sedlak and Lindsay,1968) ANOVA) followed by Dunnett’s ‘t’ test.
The glandular part of the stomach was .
homogenized in ice-cold 0.02 M EDTA. Aliquots Results
(5ml) of the homogenates were mixed in 15ml Siddha drug Kantha Chendooram showed
test tubes with 4 ml of distilled water and 1ml protective effect on indomethacin induced ulcer.
of 50% tricholoroacetic acid. The tubes were It indicates that Kantha Chendooram at the
shaken intermittently for 10 to 15 minutes and dose levels of 20 mg/kg and 40 mg/kg produced
centrifuged at 3000 r.p.m. Two ml of supernatant a significant decrease in the ulcer index
was mixed with 4ml of Tris buffer pH 8.9; (p<0.01), which is also evidenced by significant
0.1ml of 5, 5 -dithio-bis dinitro-benzoic acid increase in percentage protection from ulcers
(DTNB) was added and the sample was at the dose of 20mg/kg and 40 mg/kg (74.72 &
shaken. The absorbance was read within 5 min 82.87) respectively. The activity at both the
of addition of 5, 5 – dithio-bis 2(nitro-benzoic dose levels was comparable and equipotent as
acid) at 412 nm against a reagent blank with that of Sucralfate treated group (p<0.01).
no homogenate. (Table 1 and Figure 1)
Histological studies Figure 1.
Gastric tissue samples from each group
were fixed in 10 percent formalin for 24 hours. FIG.1 EFFECT OF S UCRALFATE, KANTHA CHENDOORAM ON ULCER INDEX
AND % ULCER PROTECTION ON INDOMETHACIN - INDUCED GAS TRIC ULCER
The formalin fixed specimens were embedded MODEL
in paraffin, sectioned (3.5-μm) and stained with 90
haematoxylin and eosin. The histological 80
sections were evaluated with light microscopy 60
Solvent Control 1m l/kg
50 Sucralfate 270 m g/kg
40 K.Chendooram 20 m g/kg
Statistical analysis 30
K.Chendooram 40 m g/kg
Results were expressed as mean ± 20
SEM. Statistical significance were determined 0
Ulcer Index % Ulcer Protection
Table 1. Effect of Sucralfate, Kantha Chendooram of Ulcer Index and Percentage Ulcer Protection
on Indomethacin Induced Ulcer Model
Group Average Treatment Dose Ulcer Positive Ulcer Percentage
Body Animals / Total Index Ulcer
Weight (g) Animals Protection
Solvent Control 1
I 243.47 6/6 40.17±3.16 -
(0.3% CMC) ml/kg
II 233.66 Sucralfate 3/6 7.0 ±0.86** 82.66
III 227.58 Kantha Chendooram 5/6 9.83±0.79** 74.72
IV 226.35 Kantha Chendooram 3/6 6.5±1.23** 82.87
Values are mean ± SEM **P<0.01 No. of animals in each group = 6
26 Vadivelan et al.
There is significant increase in mucin ameliorated ulcer index, gastro protective and
content [F (3, 20) –48.54] and NP-SH [F (3, histological changes of indomethacin induced
20) –23.97] of the gastric mucosal tissue at gastric ulcerations in rats. It is well known that
both dose levels 20mg/kg and 40 mg/kg of ulcer results from an imbalance of the interactive
Kantha Chendooram treated group. It was process of aggressive and defensive factors of
comparable and equipotent as that of sucralfate the stomach. Ulcer formation induced by
treated group (p<0.01). (Table 2) indomethacin is known to be related with
The effect of sucralfate, Kantha inhibition of cyclooxygenase that prevents
Chendooram on induction of histological lesions prostaglandin synthesis which in turn inhibits
in indomethacin induced ulcer model was shown the release of mucus, a defensive factor against
in Table 3 and Figure 2. gastrointestinal damage. (Bandyopadhyay et
Discussions Mucus secretion is a crucial factor in the
The present result suggests that pre- protection of gastric mucosa from gastric lesions
treatment with Kantha Chendooram markedly and has been regarded as an important
Table 2:Effect of Sucralfate, Kantha Chendooram of Mucus Barrier and NP-SH on Indomethacin
– Induced Ulcer Model
Group Average Treatment Dose Mucus Barrier NP-SH
Body (mcg Alcian Blue / ( mol / g tissue)
Weight (g) Glandular tissue)
I 243.47 1ml/kg 155.13 ± 3.2 0.1834 ± 0.0017
II 233.66 Sucralfate 270 mg/kg 369.93 ± 2.89** 0.6058± 0.003**
III 227.58 20 mg/kg 243.73 ± 3.46** 0.4295± 0.006**
IV 226.35 40 mg/kg 281.39 ± 1.41** 0.5666 ± 0.002**
Values are mean ± SEM *P<0.01 No. of animals in each group = 6
Table 3. Effect of Sucralfate, Kantha Chendooram on induction of Histopathological lesions in Indomethacin
– Induced Ulcer Model
Group Treatment Dose Congestion Oedema Haemorrhage Necrosis
I 1ml/kg ++ ++ ++++ ++++
II Sucralfate 270 mg/kg + + + +
III Kantha Chendooram 20 mg/kg + + ++ ++
IV Kantha Chendooram 40 mg/kg + + ++ +
+: little effect ++: appreciable effect +++: severe effect ++++: very severe effect
Effect of Kantha Chendooram in Gastric Ulcer 27
Figure 1. Histological examinations of gastric mucosal tissue sections of
control and experimental rats. (Haemotoxylin and Eosin, 20X)
Indomethacin – Induced Ulcer – Solvent Indomethacin – Induced Ulcer – Sucralfate
Control (0.3% CMC - 1 ml/kg) Treated (270 mg/kg)
Indomethacin – Induced Ulcer – Kantha Indomethacin – Induced Ulcer – Kantha
Chendooram Treated (20 mg/kg) Chendooram Treated (40 mg/kg)
defensive factor in gastric mucus barrier. A In the present study, the ulcerated region
decrease in synthesis of sulphated mucus had less mucosal NP-SH content than the intact
glycoprotein has been implicated in aetiology region in normal rats. An increase in NP-SH
of gastric ulcer. (Younan et al., 1982) content limits the production of oxygen free
The wide distribution of adherent mucus radicals and could be related with gastric
content in the gastrointestinal tract plays a protection in Indomethacin induced model. In
pivotal role in cytoprotection and repair of the the present study, rats pretreated with Kantha
gastric mucosa. (Sanyal AK et al., 1983) Chendooram showed a significant increase in
The results showed increased levels of NP-SH concentration, which might attribute to
adherent mucus content of gastric tissue its direct cytoprotection and antioxidant
pretreated with Kantha Chendooram indicating activities.
its cytoprotective action on experimentally In histological study, pretreatment with
induced gastric ulcer. Kantha Chendooram was found to preserve
28 Vadivelan et al.
the functional cytoarchitecture of the entire 3. Elango K, Suresh B, Ramesh B. R, Vadivelan
gastric mucosa. Kantha Chendooram treatment R and Sampath Kumar R : Standardization
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may be due to presence of metals (zinc, lead Pharmacology. Calcutta, Scientific Book
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In future, this work can be extended quadrangularis on gastric mucosal difensive
further for antisecretory, cytoprotective and factors in experimentally induced gastric
antioxidant studies in different ulcer models and ulcer - a comparative study with sucralfate.
J. Med Food. 7, 372-6 (2004).
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Address for correspondence: R. Vadivelan, Lecturer, Department of Pharmacology, J.S.S. College of Pharmacy,
Ooty – 643001 Tamilnadu (India). E-mail: email@example.com