Protective effect of aqueous extract of Bombax malabaricum DC on experimental models of inflammatory bowel disease in rats and mice.

Abstract

There is little evidence regarding role of B. malabaricum in the treatment of inflammatory bowel disease (IBD); though it is clinically employed as a constituent of a polyherbal preparation for IBD. To establish its role as a monotherapy for IBD, preliminary phytochemical screening of aqueous extract of B. malabaricum (AEBM) was undertaken. Subsequently, its protective effect in indomethacin and iodoacetamide induced colitis in rats (45, 90, 180, 270 mg/kg) and acetic acid induced colitis in mice (65, 130, 250, 500 mg/kg) was assessed. AEBM (270 mg/kg) in indomethacin and iodoacetamide induced colitis significantly reduced the ulcer score and myeloperoxidase (MPO) activity. AEBM/500 mg/kg dose/significantly reduced the ulcer score and MPO activity in acetic acid induced colitis. The extract (270 mg/kg in rats and 500 mg/kg in mice) was found to be comparable with prednisolone (10 mg/kg) and 5-aminosalicylic acid (5-ASA) (100 mg/kg) used as standard treatments. AEBM provided reduction in edema of the intestinal tissues, ulcer protection and lowering of MPO activity in a dose dependent manner. AEBM (500 mg/kg) significantly reduced colonic and serum TNF-alpha level when compared with the positive control in acetic acid induced colitis model. The results suggest a protective role of AEBM in IBD.

Full text

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Indian Journal of Experimental Biology
Vol. 49, May 2011, pp. 343-351






Protective effect of aqueous extract of Bombax malabaricum DC on experimental
models of inflammatory bowel disease in rats and mice
A G Jagtap* & P V Niphadkar
Department of Pharmacology, Bombay College of Pharmacy, Kalina, Mumbai 400 098, India
and
A S Phadke
Centre for Ayurveda and Panchkarma Therapy, Vashi, Navi Mumbai 400 703, India
Received 3 December 2009; revised 11 January 2011
There is little evidence regarding role of B. malabaricum in the treatment of inflammatory bowel disease (IBD); though it
is clinically employed as a constituent of a polyherbal preparation for IBD. To establish its role as a monotherapy for IBD,
preliminary phytochemical screening of aqueous extract of B. malabaricum (AEBM) was undertaken. Subsequently, its
protective effect in indomethacin and iodoacetamide induced colitis in rats (45, 90, 180, 270 mg/kg) and acetic acid induced
colitis in mice (65, 130, 250, 500 mg/kg) was assessed. AEBM (270 mg/kg) in indomethacin and iodoacetamide induced colitis
significantly reduced the ulcer score and myeloperoxidase (MPO) activity. AEBM/500 mg/kg dose/significantly reduced the
ulcer score and MPO activity in acetic acid induced colitis. The extract (270 mg/kg in rats and 500 mg/kg in mice) was found to
be comparable with prednisolone (10 mg/kg) and 5-aminosalicylic acid (5-ASA) (100 mg/kg) used as standard treatments.
AEBM provided reduction in edema of the intestinal tissues, ulcer protection and lowering of MPO activity in a dose dependent
manner. AEBM (500 mg/kg) significantly reduced colonic and serum TNF-α level when compared with the positive control in
acetic acid induced colitis model. The results suggest a protective role of AEBM in IBD.
Keywords: Bombax malabaricum, Colitis, Inflammatory bowel disease, Myeloperoxidase
Epidemiological studies suggest an increase in the
incidence and prevalence of inflammatory bowel
disease (IBD) including both Crohn’s disease (CD)
and ulcerative colitis (UC)1. The etiology of IBD
appears multifactorial. An underlying immune
dysregulation coupled with an intolerance to gut flora
seems fundamental to the pathogenesis associated
with genetic mutations or initiated by environmental
factors2.
While a cure remains still unknown, both CD and
UC can be treated with medications that induce and
maintain remission. Aminosalicylates form the
mainstay of the therapy for both CD and UC.
Antibiotics have a limited role as the disease
progresses. Steroids and immunosuppressives are
drugs of choice during the active phase of the disease
and for maintenance of remission. Newer anti-TNF-α
agents are getting approved for delivering better
results2.
Because of potential adverse events and lack of
effectiveness of standard therapies, the use of
complementary and alternative medicines, particularly
of herbal therapies, for chronic illness such as
diabetes, osteoporosis, cancer, and liver diseases is
widespread and increasing, and IBD is not excluded
from this trend3. Herbal remedies like polyherbal
extracts consisting of plants such as Aegle marmeloes,
Coriandrum sativum and individual herbal therapy
like manuka honey, etc are reported to have good
ulcer protection in IBD4,5.
Traditional Indian systems of medicines like
Siddha and Ayurveda have suggested means to
increase the body's natural resistance to disease6. A
number of Indian medicinal plants e.g. Withania
somnifera, Tinospora cordifolia, etc. known for
their ‘Rasayan’ properties have been claimed to
possess immunomodulatory activity7. Rasayana,
listed as a class in the texts of traditional Indian
medicine literature, consists of a number of plants
reputed to promote physical and mental health,
improve defense mechanisms of the body and
enhance longevity. Bombax malabaricum DC
(Sanskrit: Shalmali, Hindi: Semul, English: Red
silk cotton tree) does find a place in the traditional
rasayanas8.
____________________
*Correspondent author
Telephone: 9619058218
E-mail: jagtaparti@yahoo.com

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Preliminary phytochemical tests showed the
presence of glycosides and tannins in the root, stem
and leaf of B. malabaricum9,10. The stem, in
addition showed some alkaloids while the root
revealed the presence of proteins9,10. The isolation
of lupeol and β-sitosterol from the petroleum
extract of the stem bark has been reported9,10. The
latex contains large amounts of tannins useful as
astringent9,10. It is generally used in ‘vata’ and
‘pitta’ disorders11. Shalmali is known to be ‘Sheet’
(cold in properties-veerya-potency) and also to be a
Rasayan drug (immunomodulator drug). It is known
to be Kapha shamak, Pitta shamak and Rakta
stambhaka (haemostatic) and also known in
conquering
Rakta-Pitta-Vyadhi
disease)13. It helps in relieving the burning
sensation after burns and also does not leave the
mark11. Mocha-rasa (latex of B. malabaricum DC)
is known to possess properties such as Hima (cold),
Grahi (capable of absorbing excessive moisture or
fluid, at the same time helping in enhancing the
digestive fire – Jathragni), Snigdha (unctuous,
thereby creating smooth
preventing friction thus enhancing lubrication),
Vrushyah (helping for regeneration of tissue). Thus,
it is known for treating diseases like Pravahika
(dysentery), Atisara (diarrhea), Ama, Kapha
Shamak, Pitta Shamak and Rakta Stambhaka
(haemostatic) and anti-Dah (anti-burning)12.
In view of above information, attempts have been
made to study the effects of aqueous extract of B.
malabaricum in three different animal models of IBD,
viz. indomethacin and iodoacetamide induced colitis in
rats and acetic acid induced colitis in mice. Attempts
have also been made to elucidate the mechanism of its
ulcer protective effects by estimating myeloperoxidase
(MPO) activity and TNF-α level.

Materials and Methods
Collection and preparation of plant extractThe
dried latex (mocha-rasa) of Bombax malabaricum DC
freshly procured during the month of October was
duly authenticated by Dr. A S Phadke, Centre for
Ayurveda and Panchkarma Therapy, Vashi, Navi
Mumbai, India. The dried latex was further coarsely
powdered and extracted with distilled water. Powder
(1 kg) was macerated with 1 litre of distilled water
followed by filtration through muslin cloth. The
resultant solution was evaporated by boiling to obtain
aqueous extract of B. malabaricum (AEBM). The
yield of AEBM was 28%.
(hemorrhagic
environment and
AnimalsIn-house bred healthy adult Sprague
Dawley rats of either sex, weighing between
150-200 g and Swiss albino mice of either sex
weighing between 20-30 g, were used. Animals were
kept in temperature controlled experimental room of
animal house (23° ± 1°C) and 55 ± 10% RH. They
were subjected to 12:12 h light: dark cycle for at least 5
days prior to the study treatment ensuring their
acclimatization to the experimental conditions.
Animals were housed in standard polypropylene cages
with wire mesh top. They were fed with commercially
available rodent food pellets (Amrut rat and mice feed,
manufactured by Pranav Agro Industries Ltd., Sangli)
and water (supplied by Municipal Corporation of
Greater Mumbai). The food was withdrawn 12-18 h
prior to the induction of colitis though water. Care of
animals was taken as per the guidelines of Committee
for the Purpose of Control and Supervision of
Experiments on Animals (CPCSEA). The experiments
were performed after the approval by the Institutional
Animal Ethics Committee (IAEC) and were carried out
in accordance with the current guidelines for the care
of laboratory animals.
Drugs and chemicals usedIndomethacin (Lord
Venky’s Pvt Ltd, India) dissolved in 5% of sodium
bicarbonate (NaHCO3), acetic acid glacial AR
(Thomas Baker Pvt Ltd) diluted in distilled water,
iodoacetamide (Acros Organics) as a suspension in
1% sodium carboxy methyl cellulose (Na CMC) were
used as ulcer inducing agents. Indomethacin was
administered subcutaneously (sc) while acetic acid
and iodoacetamide were administered intrarectally
(ir). A suspension of AEBM in 1% Na CMC was
prepared. Increasing doses of suspension (45, 90, 180,
270 mg/kg, po for rats) and 65, 130, 250, 500 mg/kg,
po for mice) were used. Standard drugs used for
comparison were prednisolone (10 mg/kg, po; Lord
Venky’s Pvt Ltd, India) and 5-aminosalicylic acid
(100 mg/kg, po; Sun Pharma, India). Both of which
were given as a suspension in 1% Na CMC.
Grouping of animalsAnimals were distributed
into 8 groups of 6 animals each and received the
treatment as given in Table 1.
In indomethacin colitis model, 5 cm proximal
duodenum, 10 cm distal jejunum, 10 cm proximal
ileum, whole caecum, and 5 cm proximal colon were
isolated from all the animals for further evaluation.
In acetic acid and iodoacetamide colitis models,
5 cm distal colon and whole caecum were isolated
from all the animals for further evaluation.

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Evaluation
Morphological analysisWeight of the tissue: The
intestinal tissues (5 cm proximal ileum in indomethacin
colitis model and 5 cm distal colon in acetic acid and
iodoacetamide colitis model) were cleaned with saline,
blotted dry, placed on aluminium foil and weighed on
electronic balance. The mean weight of the intestinal
tissues of the drug and the standard treatment groups
were compared with that of the control group.
Scoring of the animal intestinal tissue: Tissues (5 cm
proximal duodenum, 10 cm distal jejunum, 10 cm
proximal ileum, whole caecum, and 5 cm proximal
colon in indomethacin colitis model; 5 cm distal colon
and whole caecum in acetic acid and iodoacetamide
colitis model) were isolated and were scored visually
for inflammation. The mean score of the drug treated
and the standard treated groups were compared with
that of the control treated group.

Scoring pattern followed is given IN Tables 2 and 3.
The final scoring is reported as sum of scores of
each part of git.
Biochemical observationsMyeloperoxidase
(MPO) assay14 and TNF-α estimation in acetic acid
induced colitis model in mice15 were carried out.
Histological analysisHistopathological analysis of
tissues for the highest dose of AEBM (270 mg/kg) was
carried out in indomethacin colitis model and in acetic
acid colitis model for 500 mg/kg close. Proximal ileum
and distal colon were isolated respectively from
indomethacin and acetic acid colitis models for
evaluation. Tissues were preserved in 10% formalin.
Microscopic studies were done by a qualified pathologist.
Table 1  Animal grouping and treatment schedule
Group

Treatment [AEBM (rats/mice in mg/kg]
po
Induction of colitis

Gr. 1
Gr. 2
Gr. 3
Gr. 4
Gr. 5
Gr. 6
Gr. 7
Gr. 8

Vehicle control
Positive control
(AEBM=45/65)
(AEBM=90/130)
(AEBM=180/250)
(AEBM=270/500)
(Standard treatment 1; Prednisolone=10)
(Standard treatment 2; 5-ASA=100)
Indomethacin (in rats, sc) Acetic acid (in mice, ir) Iodoacetamide (in rats, ir)
- -
10mg/kg 0.1ml of 7%
” ”
” ”
” ”
” ”
” ”
” ”
-
0.1ml of 1.5%
”
”
”
”
”
”
The treatment schedule was as follows:
Group

1
2
Day

1-9
1-9
8 & 9

1-9

8 & 9

1-4
Treatment

Vehicle control (1% Na CMC, po)
Vehicle control (1% Na CMC, po)
Inducing agent (in indomethacin model: 10mg/kg of indomethacin, sc; acetic acid model: 0.1ml
of 7% acetic acid, ir; iodoacetamide model: 0.1ml of 1.5% iodoacetamide, ir)
AEBM (in indomethacin and iodoacetamide models: 45, 90, 180, 270 mg/kg, po; acetic acid
model: 65, 130, 250, 500 mg/kg, po)
Inducing agent (in indomethacin model: 10 mg/kg of indomethacin, sc; acetic acid model:
0.1ml of 7% acetic acid, ir; iodoacetamide model: 0.1ml of 1.5% iodoacetamide, ir)
Standard drug (Gr. 7: 10 mg/kg prednisolone, po; Gr. 8: 100 mg/kg 5-ASA, po)
Inducing agent (in indomethacin model: 10 mg/kg of indomethacin, sc; acetic acid model:
0.1ml of 7% acetic acid, ir; iodoacetamide model: 0.1ml of 1.5% iodoacetamide, ir)
Day of
sacrifice
10
10


3-6 10

7 & 8 5
3 & 4
Table 2  Scoring pattern for rat proximal duodenum (5 cm),
distal jejunum (10 cm), proximal ileum (10 cm), and proximal
colon (5 cm) in indomethacin colitis model13
Score Observation
0
1
2
3
4
5
6
7
8
9
Table 3  Scoring pattern for rat distal colon (5 cm), mouse distal
colon (5 cm) and whole caecum (rat and mouse) in acetic acid
(in mice) and iodoacetamide (in rats) induced colitis models13
No visible change
Hyperemia at sites
Loss of mucosal integrity
Lesions having diameter 1 mm or less
Lesions having diameter 2 mm or less (no<5)
Lesions having diameter 2 mm or less (no 5-10)
Lesions having diameter 2 mm or less (no>10)
Lesions having diameter more than 2 mm (no<5)
Lesions having diameter more than 2 mm (no 5-10)
Lesions having diameter more than 2 mm (no>10)
Score Area affected (%)
0
1
2
3
4
5
6
0
0-5
5-10
10-25
25-50
50-75
75-100

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Statistical analysisStatistical analysis was done
using Student’s t test. One Way ANOVA was applied
to observe for any significant difference between the
AEBM and the standard drugs treated groups. All the
data are expressed as mean ± SD and P<0.05 was
considered to be statistically significant.

Results
During the experimental period of 10 days the
changes in weight of animals from all the groups were
comparable and none of the rats or mice was excluded
from the study.
After the induction of inflammation in the animals of
the study, colour of the stools markedly changed to
black indicative of ulceration. The stool frequency (the
need to change the animal cage) was increased after
colitis induction. This was observed in almost all the
study animal population. The behaviour of both rats and
mice was aggressive and restless after colitis induction.
Indomethacin induced colitis modelMost of the
rats in the positive control group of indomethacin
induced colitis had local peritonitis which was
reduced markedly with AEBM treatment. Rats had
transmural necrosis and inflammatory masses in small
and large bowel respectively. The mean ileum weight,
lesion score, MPO values reduced in a dose dependent
manner with AEBM treatment (Table 4). In all the three
parameters considered, the highest dose of AEBM i.e
270 mg/kg was found to be comparable with the
standard treatments. The histopathological features of
normal rat colon and caecum featured with normal intact
mucosa and with all the intestinal layers distinctly
visible with no abnormality (Fig. 1). In indomethacin
induced colitis model, transmural necrosis, edema and
inflammatory infiltrates in the mucosa were observed in
positive control rats (Fig. 1). Treatment with 270 mg/kg
of AEBM significantly attenuated the extent and
severity of the histological signs of cell damage (Fig. 1);
restoration of intestinal epithelial surface was observed
in case of standard treatments (Figs 1, 3 and 4).
Acetic acid colitis modelIn this model the
inflammation was restricted mostly to the colon. The
mean distal colonic weight, lesion score and MPO
values exhibited a significant reduction in AEBM
treated groups. AEBM (500 mg/kg) was found to be
comparable with the standard treatments. TNF-α level
Table 4  Effects of treatments on various parameters in the three models of induced colitis namely (A) indomethacin colitis in rats,
(B) acetic acid colitis in mice and (C) iodoacetamide colitis in rats
[Values are mean ± SD of 6 rats in each group]
Treatment (mg/kg, po)
[AEBM (rats/mice)]
Model Weight of the tissue
(g)
1.03 ± 0.09
0.51 ± 0.01
0.522 ± 0.019
2.9 ± 0.05
0.54 ± 0.03
2.28 ± 0.083
2.68 ± 0.24*,#
0.52 ± 0.03#
2.14 ± 0.11*,#
1.97 ± 0.12*,#
0.51 ± 0.03#
2.16 ± 0.11*,#
1.7 ± 0.7*,#
0.52 ± 0.03#
2.04 ± 0.11*,#
1.6 ± 0.04*
0.48 ± 0.03*
1.968 ± 0.1*,#
1.58 ± 0.02*
0.49 ± 0.02*
1.54 ± 0.42*
1.57 ± 0.03*
0.48 ± 0.02*
1.21 ± 0.28*
Score
(small and large intestine)
0
0
0
30.4 ± 2.3
5.6 ± 0.54
7 ± 0.7
27.1 ± 3.43# [10.85 %]
4.6 ± 1.14# [17.85%]
6 ± 0.7*,# [14.28%]
23.1 ± 1.19*,# [24.01%]
3.4 ± 1.14*,# [39.28%]
4.8 ± 0.836*,# [31.42%]
19.8 ± 1.25*,# [34.86 %]
3 ± 1.22*,# [46.42%]
3.2 ± 0.836*,# [54.28%]
14.4 ± 1.78* [52.63 %]
2 ± 1.09* [64.28%]
2.4 ± 0.894*,# [65.71%]
13.7 ± 1.15* [54.93 %]
1 ± 0.7* [84.14%]
1.6 ± 0.547* [77.14%]
13 ± 1.62* [57.23 %]
0.8 ± 0.44* [85.72%]
1.4 ± 0.547* [80%]
MPO (U/g)
(in 10 cm proximal ileum)
5.04 ± 0.25
1.19 ± 0.57
15.37 ± 0.766
44.992 ± 1.86
7.4 ± 1.09
41.954 ± 3.405
23.71 ± 3.07*,#
6.7 ± 0.94,#
38.19 ± 1.885*,#
21.876 ± 3.4*,#
5.7 ± 1.01*,#
35.49 ± 2.316*
18.522 ± 2.30*,#
5.3 ± 0.99*,#
27.7 ± 2.57*
10.25 ± 0.97*,#
3.46 ± 1.41*
23.29 ± 1.89*
6.09 ± 0.93*
2.88 ± 0.63*
19.62 ± 2.568*
5.57 ± 0.74*
2.6 ± 0.64*
18.83 ± 1.077*
Vehicle control
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
Positive control
1st dose of AEBM
(45/65)
2nd dose of AEBM
(90/130)
3rd dose of AEBM
(180/250)
4th dose of AEBM
(270/500)
Prednisolone = 10
5-ASA = 100
P<0.05 significant with respect to *positive control; standard treatments (Predrisolone and 5-ASA) One way ANOVA followed by
Bonferroni’s multiple comparison test

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estimated for highest effective dose of AEBM 500
mg/kg (serum/colonic) was found to be reduced
significantly compared to the respective positive control
groups (Fig. 2). The colonic TNF-α level was found to
be comparable with the standard drugs treated group.
Iodoacetamide induced colitisInflammation was
limited to the distal part of the colon. The mean distal
colonic tissue weight, lesion score, MPO values after
AEBM treatment were found to be significantly lower
than the positive control group and showed dose


Fig. 1
showing massive necrosis and inflammatory infiltrate; D: AEBM (270 mg/kg) pretreatment attenuated the extent and severity of damage;
e and f: prednisolone and 5-ASA treated sections with much improved histology (M: mucosa, S: submucosa, ML: muscle layer, V: villi,
I: inflammatory infiltrate, N: necrosis, VC: villi collapsed, MI: mild inflammation)]

 Histological sections of rat ileum in indomethacin induced colitis model[ Normal intact mucosa; b and c: positive control

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dependent effect with AEBM (270 mg/kg) being
comparable to the standard treatments.

Discussion
Over the last few decades, inflammatory bowel
disease including both Crohn’s disease and ulcerative
colitis has claimed many lives globally. New insights
in the treatment strategies provide hope that a curative
measure would be developed against IBD1.
In the present study three different animal models
were employed in an attempt to diversify the animal
models to mimic human
Indomethacin16,17 and iodoacetamide18 induced colitis
in rats elicit features resembling human CD, whereas
acetic acid induced colitis in mice resembles human
UC19. Based on the consistency of observations of the
ulcerated animals, parts of intestine were isolated for
scoring and assay purposes.
Not much data are available regarding the use
of Bombax malabaricum as a monotherapy for IBD.
intestinal disease.
B. malabaricum (Shalmali) is a Rasayan drug mentioned
in authentic scripts12. Further, there are references
available on Rasayan drugs with regard to their
immunomodulatory/anti-oxidant activities that helped us
identify the drug for this study6,7. The clinical experience
of one of the authors, Dr. Ashish Phadke revealed that
Shalmali when administered by different routes
(oral and enema) lessened the severity of the
manifestations of IBD (Phadke, unpublished data).
Therefore it was thought of establishing and elucidating
its role and mechanism involved in reducing
inflammation in IBD as per the principles of reverse
pharmacology20.
Non-steroidal anti-inflammatory drugs (NSAIDs) like
indomethacin produce acute ileitis. NSAID induced
enteropathy is one of the most interesting models of CD
since it shares the same early modification observed in
human CD, like increased intestinal permeability
followed by intestinal
inhibits cyclo-oxygenase (COX) enzyme thus inhibiting
lesions17. Indomethacin

Fig. 2 TNF-α estimation in intestinal tissues and serum in acetic acid induced colitis model in mice [Values are mean ± SD of 3 mice in
each group P<0.05 significant with respect to * positive control; # standard treatments (Prednisolone and 5-ASA) (One way ANOVA
followed by Bonferroni’s multiple comparison test)]



Fig. 3 Photographs of indomethacin induced colitis in rat ileum [The number and severity of ulcers seen in Grp 2 significantly reduces
with increasing doses of AEBM (groups 3-6)].

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the formation of prostaglandins (PGs). As a result, the
protective action of PGs is lost which is a predisposing
factor for gastric ulceration and erosion16,17. In the
present study, indomethacin produced gross ulceration
in the small and large bowel with skip lesions. In all the
animals of this model,
(morphological and histological) was observed in the
proximal ileum. Caecum was also affected severely
exhibiting gross ulceration or in some cases patches of
inflammation (Figs 3 and 4). From morphological
analysis like edema measurement and lesion scoring of
tissues it can be concluded that AEBM was effective in
reducing the edematous growth in IBD. The stool colour
and frequency was observed to be returning to normalcy
in animals treated with higher doses of AEBM.
Reduction of MPO levels showed anti-oxidant potential
of AEBM. Histopathological findings clearly show the
presence of inflammatory infiltrate cells (granulocytes
and macrophages) in the mucosa and severely collapsed
villi in the ileum of positive control group whereas it
markedly improved after AEBM treatment (Fig 2).
UC is a chronically recurrent inflammatory bowel
disease of unknown origin. Oxidative stress has been
implicated in the pathogenesis of UC in experimental
animals21,22. Epithelial injury in acetic acid induced
UC model in mice is caused by the entry of lipid
soluble (protonated) form of acetic acid into
epithelium which dissociates to liberate protons into
intracellular space21. The distal colonic weight
reduced with AEBM treatment indicating its potential
to reduce edema. AEBM was also effective in
reducing ulceration (lesion score) of the colon in UC
induced animals. Colitis could also be caused by the
activated neutrophils which pass out of the circulation
and enter the inflamed mucosa and submucosa during
acute inflammation leading to overproduction of
consistent ulceration
reactive oxygen species that leads to further intestinal
injury22. AEBM treatment led to a dose dependent
decrease in MPO values (index of neutrophil
recruitment) which is evocative of its anti-oxidant
property. Histopathological analysis of highest dose
of AEBM treatment revealed that the extent of
damage produced by acetic acid was reduced
significantly. The present results also showed that the
proinflammatory cytokine TNF-α production was
increased in colonic mucosa after acetic acid
instillation. Cytokines are key signals in the intestinal
immune system, and are known to participate in the
disruption of the so-called normal state of controlled
inflammation23. TNF-α is one of the most important
cytokine participating in the inflammatory process of
patients with IBD24,25. TNF-α may amplify mucosal
inflammation by a number of different mechanisms. It
induces the production of other cytokines including
adhesion molecules, arachidonic acid metabolites, and
activation of immune and non-immune cells.
Antibodies of avian TNF-α effectively treated
inflammatory bowel diseases in rats26 and in
humans27. TNF-α expression in human macrophages
was discovered in the colonic tissue in both patients
with CD and UC28 and serum levels of TNF-α
correlate with clinical and laboratory indices of
intestinal disease activity29. So, it was thought
worthwhile to assess the TNF-α levels in mice (acetic
acid colitis model) to ascertain its role in UC. TNF-α
level in serum/colonic samples reduced significantly
as compared to their respective positive control group.
This anti-inflammatory property could be a probable
result of its anti-oxidant property or Nuclear Factor-
κB inhibition. NF-κB is activated in patients with
inflammatory bowel disease30 and in rats with
trinitrobenzene sulfonic acid-induced colitis31.


Fig. 4 Photographs of indomethacin induced colitis in rat caecum [The ulcerated patches on the caecum (Group 2) is visibly reduced
with AEBM treatment (Groups 3-6)].

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Iodoacetamide colitis follows a biochemical
mechanism of action. Endogenous sulfhydryl
compounds have an important role in maintaining
mucosal integrity in the GIT18. The injury induced
by iodoacetamide is maximal when assessed both
macroscopically and histologically up to seven days
after its exposure to the colonic mucosa32.
Macroscopical analysis like lesion scores of the
distal colon and edema (weight of the colon) was
found to be reducing with increasing doses of
AEBM. The colonic MPO activity was found to be
stimulated in positive control treated group,
indicating the contribution of granulocytes to the
inflammatory cascade. AEBM treated tissues
exhibited reduction in MPO activity in a dose
dependent manner.
Accumulating data from three different animal
models producing colitis by way of different
mechanisms indicate and reconfirm the multifactorial
etiology of IBD. Based on the availability of animals,
rats and mice were used in the study accordingly.
Only one strain of rat (Sprague Dawley) and mice
(Swiss albino) was used.
Thus, the present study supports the proposition of
above mentioned ayurvedic literature that Shalmali
(B. malabaricum) possesses
properties as well as antioxidant properties.
The mocha-rasa of Shalmali is known to contain
large amounts of tannic and gallic acids11 acting as
astringents which precipitate proteins helpful in
restoring the damaged epithelial mucosal lining of
the ulcerated mucosa. Also, the animal population
with AEBM pretreatment exhibited significantly
lower edema, macroscopic score and MPO value
which reveal that AEBM does possess a protective
effect against the known colitis inducing agents.
Shalmali which has properties like Sheet (cold in
property-relieve burning sensation of ulceration),
Kashaya (Astringent), haemostatic, usefulness in
Pravahika and also possessing quality of Rasayan
drug, coupled with the findings. The present study
revalidates its use as a drug of choice in treatment of
IBD.
Further characterization of the plant extract could
lead to interesting findings which can provide more
accurate details about the mechanisms by which
Shalmali acts as a protective agent. Thus,
B. malabaricum (shalmali) may prove to be an
important and effective indigenous drug in future for
the treatment of IBD.
anti-inflammatory
Acknowledgement
Thanks are due to Dr R D Gupte (Piramal Life
Sciences) for help in TNF-estimation and Dr Sanjay
Pawar for histopathological analysis of the samples.

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