The cytoprotective consequence of four preparations variables ( Batch A, Batch B, Batch C and Batch D ) of snail mucin and Tagamet were evaluated utilizing three animate being ( albino rats ) theoretical accounts ( Indocin, histamine and stress-induced ulcer theoretical accounts ) . Results show that the cytoprotective consequence of the preparation variables increased with addition in the concentration of snail mucin infusion. The negative control which was Tween 85 had no important consequence on the ulcer ( p & lt ; 0.05 ) . This suggests that a combination of snail mucin and Tagamet in drug bringing may hold some curative importance and should be decently harnessed.
Keywords: peptic ulcer ; snail mucin ; Tagamet ; cytoprotective ; carnal theoretical accounts
Peptic ulcer is a breach 1 or a sore 2 in the liner ( mucous membrane ) of the digestive piece of land produced by digestion of the mucous membrane by acid-pepsin3. Peptic ulceration develops in the epithelial lined surfaces exposed to the acerb secernment of the stomachic secretory organs. The sites most frequently affected are the tummy itself, the duodenum bulb and the distal portion of the oesophagus4.
It is known that non-steroidal anti-inflammatory agents are capable of doing a assortment of acute stomachic lesions, which may climax in stomachic erodings or even blunt ulceration. Sometimes drug induced mucosal harm is accompanied by extended bleeding into the mucosa5. Histamine, by exciting the cAMP-dependent tract, leads to increased stomachic secernment 6.The function of emphasis is controversial7. Wilson and Waugh, 2000 8 reported that with emphasis, there is addition in secernment of norepinephrine and epinephrine endocrines that cause bottleneck of the blood vass providing the alimental canal.
The presence or absence of peptic ulcer is determined by the delicate interplay between aggressive factors ( secreted gastric acid and pepsin ) and defensive factors ( mucosal opposition ) . Peptic ulcer is produced when the aggressive effects of acid-pepsin dominate the protective effects of gastric or duodenal mucosal resistance5, 9.
Mucus is a extremely syrupy fluid secreted by mucose membranes and secretory organs, dwelling of mucin, leukocytes, inorganic salts, H2O and epithelial cells10. It lubricates the walls of the piece of land and protects them from digestive enzymes11. It is extremely biocompatible, non toxic and easy biodegradable. In future, it may play a cardinal function in the pharmaceutical industry as a drug bringing agent12.
H2-receptor adversaries have been observed to protect experimental animate beings from stomachic ulceration induced by emphasis, pyloric ligation, acetylsalicylic acid, H2- receptor agonist, or cholimimetics but with no consistence on stomachic emptying rate13.
Although several chemical challenges still need to be met in this country of research, it is sensible to reason ( ? ? expect ) that with the coming of drug-mucin combinations, the depredations of gastric and duodenal ulcers will be reduced.
[ Explain clearly why the drug mucin combination is expected to increase antiulcerative belongings of the exemplary drug. ]
The following were procured from their local providers and used without farther purification ; Tagamet, Indocin, histamine, snail mucin infusion and distilled H2O.
The elephantine African snails, Archachatina marginata, Fam. Arionidae were procured from Ibagwa-Nkwo market in Nsukka zone of Enugu State. White albino rats ( 110-180g ) of either sex, obtained from the carnal house of the Department of Pharmacology and Toxicology, University of Nigeria, Nsukka, were used for the survey. While in the animate being house, they had free entree to nutrient and H2O and were maintained under standard conditions. All carnal handling and experiments were conducted following the guidelines stipulated by University of Nigeria Research Ethics Committee on Animal Handling and Use.
Extraction of snail mucin
After procurance, the shells of 50 elephantine African land snails were knocked unfastened at the vertex and a spirally coiled rod inserted to take the heavy organic structure. This heavy portion was placed in 250 milliliter of H2O and washed each clip until the mucin was thoroughly washed off. The mucin was pooled together and lyophilized manually. It was later dried in air. The flakes were pulverized and stored in an airtight container until used14.
Preparation of exogenic mucin and Tagamet alloies
Binary alloies of Tagamet and exogenic mucin were assorted exhaustively in the ratios of 1:1 ; 1:2 ; 1:3 ; 1:4 severally. Table 1 shows the drug-mucin preparation variables.
Table 1: combination ratios of drug: mucin
Cimetidine ( milligram )
Snail Mucin ( milligram )
Ulcer experiments in albino rats
White albino rats ( 110-180g ) of either sex, obtained from the carnal house of the Department of Pharmacology and Toxicology, University of Nigeria, Nsukka, were used for the survey. Three theoretical accounts of bring oning experimental stomachic ulcers were used to measure the anti-ulcer activity of the preparation batches. The theoretical accounts were: Indocin, histamine and stress- induced ulcer theoretical accounts. The method followed was as described by Akah and Nwafor 15.
Indomethacin induced ulcer
Rats were fasted for 18 H prior to the beginning of the experiment and there were 24 rats for each theoretical account. They were divided into six groups of four rats each. The first group ( A ) received cimetidine merely. It served as positive control at a dosage of 500mg/kg. The 2nd group ( B ) received 1:1 combination of Tagamet and exogenic snail mucin. The 3rd group ( C ) received combination of Tagamet and snail mucin at a ratio of 1:2. The 4th group ( D ) received combination of Tagamet and exogenic snail mucin at a ratio of 1:3 and the 5th group ( E ) received Tagamet and exogenic snail mucin at a ratio of 1:4. The 6th group ( F ) group received 3 % tween 85. [ Explain the ground for including Tween 85 in the survey and why it has been called negative control in the abstract ] Thirty proceedingss subsequently, ulcer was induced by administrating 30mg/kg organic structure weight of Indocin ( dissolved in 3 % tween 85 ) to the different groups of animate beings severally. All disposals were by the unwritten path.
After 8 H for indomethacin- induced ulcer theoretical account, the animate beings were killed and the tummies removed and opened along the greater curvature. The tummies were rinsed under a watercourse of H2O and pinned level on a corkboard. The tummies were observed with a manus lens ( x10 ) .
Histamine induced ulcer
Ulcer was induced in experimental animate beings utilizing 2mg/kg of histamine disposal orally to the animate beings. The process for indomethacin-induced ulcer was followed. Ulcer was observed every bit at the junction between the antrum and fundus of the tummy. The ulcer was viewed and counted, each given a badness evaluation as follows harmonizing to Main and Whittle 16: & lt ; 1 millimeter = 1 ; & gt ; 1 & lt ; 2 millimeter = 2 ; & gt ; 2mm = 3. The overall sum divided by a factor of 10 was designated as the ulcer index ( ui ) for that tummy.
Based on their strength, the ulcer can besides be given tonss as follows: 0 = no ulcer, 1 = superficial mucosal eroding, 2 = deep ulcer, 3 = perforated ulcers 17.
Ulcer index = arithmetic mean of strength in a group + figure of ulcer positive animate beings / entire figure of animals18
Cold restrained emphasis induced ulcer
The method demonstrated by Suzuki 19 was adopted to bring on ulcer by emphasis in the research lab animate beings. The Tagamet aa‚¬ ” mucin ratios were administered 30 min prior to subjecting the animate beings to emphasize. The animate beings were placed in a reticent coop and the coops were placed at icebox temperature for 3h. The animate beings were sacrificed and so the ulcer protection was determined20, 21.
Percentage ulcer protection = 1- ( ulcer index for trial agent/ulcer index for negative control ) .
Gastric motility trial in albino rats
Twenty rats of either sex were indiscriminately divided into five animate beings per group. The animate beings were starved for 24 H prior to the experiment, but had free entree to H2O. One group received Tween 85 ( 5 ml/kg ) , while the staying four groups received the different doses of the alloies. All disposals were made by the unwritten path. Five proceedingss after drug disposal, 0.5ml of a 5 % wood coal suspension in 10 % aqueous solution of tragacanth pulverization was administered to each animate being. The animate beings were sacrificed 30 min subsequently and the venters opened. The per centum distance of the little bowel ( from pylorus to caecum ) traveled by the wood coal stopper in the treated animate beings were determined15
The consequences were analyzed utilizing pupils t-test and were regarded as important at P & lt ; 0.05.
RESULTS AND DISCUSSION
Consequence of ulcer experiment
Indomethacin-induced ulcer was significantly ( p & lt ; 0.05 ) protected by combinations of Tagamet and snail mucin. It showed a dosage related dependent protection with the least ulcer index from 1:3 and 1:4 ; Tagamet: snail mucin combination severally. Histamine -induced ulcer besides showed dose-related protection of the rats. The stress-induced ulcer showed higher protection from the 1:1 combination of drug: Tagamet ( intending non clear ) than from the drug-snail mucin combination. There was no protection from the negative control which was tween 85. The ends of therapy for ulcers are to alleviate hurting, to advance complete healing, to forestall reoccurrence and to forestall the development of complications22. Although, the addition of Tagamet: mucin combinations decresed ulcer indices in two of the three theoretical accounts, better protection appears to be shown against indomethacin-induced ulcer proposing better cytoprotective mechanism of action. The combination did non demo important protection in the stress-induced ulcer particularly with the higher dose combinations.
The consequence of the wood coal meal trial is shown in Table 3. The disposal of increasing concentration of mucin significantly reduced the wood coal repast theodolite. Since the little bowel is the primary site of drug soaking up, the longer the abode clip in this part the greater is the possible for efficient drug soaking up presuming that the drug is stable in the enteric fluid and does non respond with endogenous stuffs to organize hapless absorbable aa‚¬E?complexesaa‚¬a„? . The abode clip in the little bowel as determined by enteric motility, 23 may be an of import factor with regard to drug bioavailability. The longer a drug is in contact with the soaking up site ( s ) the greater the sum of drug absorbed24. In ulcer patients, decrease in gut motility helps to better the ulcer hurting and rush the healing of ulcer wounds25.
Table 2a: Indocin and histamne induced ulcer
Number of animate beings used
ulcer mean index for Indocin
ulcer mean index for histamine
26.67 + 1.43
8.67 + 1.77
5.67 + 0.65
5.67 + 2.19
11.33 + 0.35
7.33 + 2.72
3.33 + 2.03
4.00 + 0.58
3.33 + 2.03
4.00 + 0.58
Table 2b: Stress induced ulcer
Number of animate beings used
Table 3: Consequence of trial agents on GI motility
Dose ( mg/kg )
% distance travelled
Cimetidine: mucin 1:1
Cimetidine: mucin 1:2
18.14 + 1.0
Cimetidine: mucin 1:3
Cimetidine: mucin 1:4
Significant ( P & lt ; 0.05 ) relation to 3 % tween 85