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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
                                                     (g)]
      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
                                                          70
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
                                                          10
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
                                                  270
    II        233.66           Sucralfate                              3/6                 7.0 ±0.86**                  82.66
                                                 mg/kg
                                                  20
   III        227.58      Kantha Chendooram                            5/6                9.83±0.79**                   74.72
                                                 mg/kg
                                                  40
   IV         226.35      Kantha Chendooram                            3/6                 6.5±1.23**                   82.87
                                                 mg/kg
  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
                                                                     al., 2000)
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)
                                  Solvent Control
      I           243.47                                     1ml/kg             155.13 ± 3.2               0.1834 ± 0.0017
                                   (0.3% CMC)
     II           233.66                Sucralfate         270 mg/kg          369.93 ± 2.89**              0.6058± 0.003**
                                         Kantha
     III          227.58                                   20 mg/kg           243.73 ± 3.46**              0.4295± 0.006**
                                       Chendooram
                                         Kantha
     IV           226.35                                   40 mg/kg           281.39 ± 1.41**             0.5666 ± 0.002**
                                       Chendooram
             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
                        Solvent Control
         I                                       1ml/kg               ++            ++             ++++              ++++
                         (0.3% CMC)
      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
showed not only the maintenance but also the                        of Kantha Chendooram. Ancient Science of
regeneration of gastric mucosa in the damaged                       life. Vol: XXVI (1&3), 89-91 (2006).
                                                           4.       Formulary of Siddha Medicines : The Indian
regions. These findings confirm the
                                                                    Medical Practitioners. Co-operative Pharmacy
cytoprotective nature of Kantha Chendooram.                         and Stores Ltd., Publication, Chennai. IV
(Jainu M and Devi CSS, 2004)                                        edition 215-515 (1993).
       The antiulcer activity of the Siddha drug           5.       Ghosh M. N : Fundamentals of Experimental
may be due to presence of metals (zinc, lead                        Pharmacology. Calcutta, Scientific Book
and mercury) and alkaloid present in it. (Elango                    Agency, II edition, 5 & 155 (1984).
K et al., 2006)                                            6.       Jainu M and Devi C. S. S : Effect of Cissus
       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).
safety profile of the drug, so far not attempted.
                                                           7.       Kulkarni S.K and Goel R. K : Gastric
                                                                    antiulcer activity UL-409 in rats. Indian J.
Acknowledgements                                                    Exp Biol. 34, 683-688 (1996).
       The authors are grateful to the Director,           8.       Njar V. C. O, Adesanwo J. K, Makinde
Microlaboratories, Coimbatore for the                               J.M and Jaiwo O. B : Antiulcer activity of
histopathology studies, Technical Officer,                          the stem bark extract of Entandrophargma
National Metallurgical Laboratory, Chennai for                      angolenses. Phytother Res. 8, 44-8 (1994).
the analytical report of the Siddha drug and the           9.       Sanyal A. K, Mitra P. K and Goel R. K : A
Management, J.S.S. College of Pharmacy, Ooty,                       modified method to estimate dissolved
India for providing the necessary infrastructure                    mucosubstance in gastric juice. Indian J Exp
                                                                    Biol. 21: 78-80 (1983).
to carry out this research work in successful
                                                           10.      Sedlak J and Lindsay R : Estimation of total,
manner.                                                             protein bound sulphydryl groups in tissue
                  References                                        with Ellmans reagent. Anal Biochem. 25,
1.         Asuzu I. U and Omu U : Antiulcer activity of             192- 5 (1968).
           the ethanol extract of Combretum                11.      Suleyman H, Demirezer L.O and
           dolichopetalum. Int. J. Crude Drugs Res.                 Buyukokurolu M. E : Antiulcerogenic effect
           28 (1): 27-32 (1990).                                    of Ippophae rhamnoides L. Phytother Res.
2.         Bandyopadhyay S. K and Pakrashi S. C                     15: 625-7 (2001).
           and Pakrashi A : The role of antioxidant        12.      Younan F, Person J, Allen A and Enables C :
           activity of Phyllanthus emblica fruits on                Changes in the structure of the mucus gel in
           prevention from indomethacin-induced                     the mucosal surface of the stomach in
           gastric ulcer. Ethnopharmacol. 70: 171-6                 association with peptic ulcer disease.
           (2000).                                                  Gastroenterology. 82, 827-31 (1982).




Address for correspondence: R. Vadivelan, Lecturer, Department of Pharmacology, J.S.S. College of Pharmacy,
Ooty – 643001 Tamilnadu (India). E-mail: velcology@yahoo.com
044_2007

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Anti-ulcer effects of Siddha drug Kantha Chendooram

  • 1. 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
  • 2. 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 (g)] 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
  • 3. 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 70 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 10 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 270 II 233.66 Sucralfate 3/6 7.0 ±0.86** 82.66 mg/kg 20 III 227.58 Kantha Chendooram 5/6 9.83±0.79** 74.72 mg/kg 40 IV 226.35 Kantha Chendooram 3/6 6.5±1.23** 82.87 mg/kg Values are mean ± SEM **P<0.01 No. of animals in each group = 6
  • 4. 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 al., 2000) 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) Solvent Control I 243.47 1ml/kg 155.13 ± 3.2 0.1834 ± 0.0017 (0.3% CMC) II 233.66 Sucralfate 270 mg/kg 369.93 ± 2.89** 0.6058± 0.003** Kantha III 227.58 20 mg/kg 243.73 ± 3.46** 0.4295± 0.006** Chendooram Kantha IV 226.35 40 mg/kg 281.39 ± 1.41** 0.5666 ± 0.002** Chendooram 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 Solvent Control I 1ml/kg ++ ++ ++++ ++++ (0.3% CMC) 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
  • 5. 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
  • 6. 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 showed not only the maintenance but also the of Kantha Chendooram. Ancient Science of regeneration of gastric mucosa in the damaged life. Vol: XXVI (1&3), 89-91 (2006). 4. Formulary of Siddha Medicines : The Indian regions. These findings confirm the Medical Practitioners. Co-operative Pharmacy cytoprotective nature of Kantha Chendooram. and Stores Ltd., Publication, Chennai. IV (Jainu M and Devi CSS, 2004) edition 215-515 (1993). The antiulcer activity of the Siddha drug 5. Ghosh M. N : Fundamentals of Experimental may be due to presence of metals (zinc, lead Pharmacology. Calcutta, Scientific Book and mercury) and alkaloid present in it. (Elango Agency, II edition, 5 & 155 (1984). K et al., 2006) 6. Jainu M and Devi C. S. S : Effect of Cissus 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). safety profile of the drug, so far not attempted. 7. Kulkarni S.K and Goel R. K : Gastric antiulcer activity UL-409 in rats. Indian J. Acknowledgements Exp Biol. 34, 683-688 (1996). The authors are grateful to the Director, 8. Njar V. C. O, Adesanwo J. K, Makinde Microlaboratories, Coimbatore for the J.M and Jaiwo O. B : Antiulcer activity of histopathology studies, Technical Officer, the stem bark extract of Entandrophargma National Metallurgical Laboratory, Chennai for angolenses. Phytother Res. 8, 44-8 (1994). the analytical report of the Siddha drug and the 9. Sanyal A. K, Mitra P. K and Goel R. K : A Management, J.S.S. College of Pharmacy, Ooty, modified method to estimate dissolved India for providing the necessary infrastructure mucosubstance in gastric juice. Indian J Exp Biol. 21: 78-80 (1983). to carry out this research work in successful 10. Sedlak J and Lindsay R : Estimation of total, manner. protein bound sulphydryl groups in tissue References with Ellmans reagent. Anal Biochem. 25, 1. Asuzu I. U and Omu U : Antiulcer activity of 192- 5 (1968). the ethanol extract of Combretum 11. Suleyman H, Demirezer L.O and dolichopetalum. Int. J. Crude Drugs Res. Buyukokurolu M. E : Antiulcerogenic effect 28 (1): 27-32 (1990). of Ippophae rhamnoides L. Phytother Res. 2. Bandyopadhyay S. K and Pakrashi S. C 15: 625-7 (2001). and Pakrashi A : The role of antioxidant 12. Younan F, Person J, Allen A and Enables C : activity of Phyllanthus emblica fruits on Changes in the structure of the mucus gel in prevention from indomethacin-induced the mucosal surface of the stomach in gastric ulcer. Ethnopharmacol. 70: 171-6 association with peptic ulcer disease. (2000). Gastroenterology. 82, 827-31 (1982). Address for correspondence: R. Vadivelan, Lecturer, Department of Pharmacology, J.S.S. College of Pharmacy, Ooty – 643001 Tamilnadu (India). E-mail: velcology@yahoo.com 044_2007