Effect Of Siddha Drug (Kantha Chendooram) On


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Effect Of Siddha Drug (Kantha Chendooram) On

  1. 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. 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. 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. 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. 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. 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