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 Table of Contents  
ORIGINAL ARTICLE
Year : 2017  |  Volume : 20  |  Issue : 2  |  Page : 189-195

Healing effect of Allium sativum on induced upper gastrointestinal tract injury in albino Wistar rats


1 Department of Medical Laboratory Sciences, University of Benin, Benin City, Edo State, Nigeria
2 Department of Anatomy, University of Benin, Benin City, Edo State, Nigeria

Date of Web Publication14-Nov-2017

Correspondence Address:
Frederick Olusegun Akinbo
Department of Medical Laboratory Sciences, University of Benin, Benin City, Edo State
Nigeria
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DOI: 10.4103/tjmr.tjmr_13_16

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  Abstract 

Background: Peptic ulcer is regarded as a chronic disease that impairs the quality of life and has been linked to increased morbidity and mortality. This study was conducted to determine the ameliorative effect of Allium sativum on induced gastrointestinal tract injury in albino Wistar rats. Materials and Methods: A. sativum was purchased from vegetable market and authenticated in the Department of Pharmacognosis of University of Benin, Benin City, Edo State. A total of 24 albino Wistar rats of both sexes weighing between 120 g and 150 g were used. The rats were randomly shared into six study groups: two control groups and four treated groups (four rats of each). Group A consisted of rats given 1 ml of normal saline and sacrifi ced after 6 h. Group B were rats given 20 mg/kg of indomethacin (n = 4) and sacrifi ced after 6 h. Group C consisted of four rats given 20 mg/kg of indomethacin (n = 4) and treated with aqueous extract of A. sativum (100 mg/kg) body weight, twice daily for 8 days and thereafter sacrificed (n = 4). Group D were given 20 mg/kg of indomethacin (n = 4) and then treated with aqueous extract of A. sativum (200 mg/kg) body weight, twice daily for 8 days and sacrifi ced (n = 4). Group E consisted of four rats given 20 mg/kg of indomethacin (n = 4) and treated with aqueous extract of A. sativum (300 mg/kg) body weight, twice daily for 8 days and sacrifi ced (n = 4). Group F consisted of four rats given 20 mg/kg of indomethacin (n = 4) and afterward treated with cimetidine (50 mg/kg) body weight twice daily for 8 days and sacrificed. Results: A. sativum exhibited better healing activities at 100 mg/kg dosage than 300 mg/kg while cimetidine produced fairly normal mucosa in the stomach and mild mucosa sloughing in the duodenum of the Wistar rats. The 200 mg/kg and the 300 mg/kg doses of A. sativum presented with similar results. Conclusion: Suffice to say that garlic extract is better than cimetidine in the treatment of gastric and duodenal ulcer. Further studies into the antiulcer activity of A. sativum are advocated.

Keywords: Albino rats, Allium sativum, gastrointestinal tract injury


How to cite this article:
Akinbo FO, Eze G. Healing effect of Allium sativum on induced upper gastrointestinal tract injury in albino Wistar rats. Trop J Med Res 2017;20:189-95

How to cite this URL:
Akinbo FO, Eze G. Healing effect of Allium sativum on induced upper gastrointestinal tract injury in albino Wistar rats. Trop J Med Res [serial online] 2017 [cited 2018 Dec 14];20:189-95. Available from: http://www.tjmrjournal.org/text.asp?2017/20/2/189/218206


  Introduction Top


Peptic ulcer is regarded as a chronic disease that impairs the quality of life and has been linked to increased morbidity and mortality.[1] An estimated 4 million people are affected by peptic ulcer disease yearly all around the world.[2] Of this estimate, complications are encountered in 10%–20% of these patients and 2%–14% of the ulcers will perforate.[3] Although rare, perforated peptic ulcers could be life-threatening with mortality varying from 10% to 40%.[4],[5]

The etiology of gastric ulcers is influenced by various aggressive factors which may include bile acids, acid-pepsin secretion, Helicobacter pylori infection; protective factors including parietal cell tight junction, mucosal barrier, mucus secretion, blood flow, cellular regeneration, and endogenous protective agents, namely – prostaglandins and epidermal growth factors.[6] Other causes of this disease may be stress, smoking, nutritional deficiencies, and ingestion of nonsteroidal anti-inflammatory drugs.[7],[8]

The most known common ulcer symptom is a burning pain in the abdomen between the breastbone and the navel occurring between meals and in the early hours of the day spanning a few minutes to hours. The uncommon ulcer symptoms are anemia, bloating, belching, nausea and vomiting, blood-tinged vomit, poor appetite, loss of weight, and feeling tired and weak.[9] People with ulcers may experience serious complications such as bleeding, perforation, and narrowing and obstruction in the posterior end of the stomach and the duodenum.[10]

Allium sativum (commonly known as garlic) belongs to the Liliaceae (onion) family and is one of the nature's most versatile medicinal plants. It has been in use for over thousands of years for a wide range of health conditions and traditional dietary supplement for diabetes in Asia, Europe, and the Middle East.[11] Fresh garlic has been found to be of some interesting biological and pharmacological activities including antifungal and antibacterial effects.[12] The garlic plant's bulb is the most commonly used part of the plant as it contains 84.09% water, 13.38% organic matter, and 1.53% inorganic matter, while the leaves are 87.14% water, 11.27% organic matter, and 1.59% inorganic matter.[13] The phytochemicals for the strong flavor of garlic are produced when the cells of the plants are damaged through chopping, chewing, or crushing, thereby causing the breakdown of several sulfur-containing compounds stored in the cell fluids. The resultant products (compounds) are responsible for the strong or hot taste and smell of garlic.[13]

The drugs that are currently available for the treatment of peptic ulcers are not free from side effects. Against this background, this study was conducted to determine the ameliorative effect of A. sativum on induced gastrointestinal tract injury in albino Wistar rats.


  Materials and Methods Top


Plant collection and extraction

A. sativum was purchased from the vegetable market at Airport Road, Benin City, Edo State. The plant sample was authenticated in the Department of Pharmacognosis of University of Benin, Benin City, Edo State. About 250 g of A. sativum plant was washed in water and crushed to semi-liquid form using a mechanical blender. About 500 ml of distilled water was added and shaken by mechanical shaker for 4 h and left overnight. The mixture was sieved with a Whatman (number 1) filter paper and filtrate allowed to evaporate in an oven at 40°C. The resultant residue was resuspended and then used as the aqueous extract. Of this powder, 100, 200, and 300 mg/kg body weight were prepared in 100 ml of distilled water before use.[14] Approximately 1 ml of the extract was administered to the rats with the help of a gavage while giving them their normal feed (livestock grower's marsh) and drinking water ad libitum.

Animal

A total of 24 albino Wistar rats of both sexes weighing between 120 g and 150 g were used. The animals were randomly divided into groups and were kept under standard conditions of temperature (23°C ± 2°C) and humidity. The rats were kept for 2 weeks in the animal house at the Faculty of Life Sciences, University of Benin, before the experiment to acclimatize. They were maintained on unrestricted supplies of food and water. Food was withdrawn 24 h before the inducement of ulcer on the rats while water was maintained. The rats were randomly shared into six study groups: two control groups and four treated groups (four rats of each). In handling the rats, the National and Institutional Guidelines for the Protection of Animal were followed.

Induction of ulcer

Following 24 h of fasting, 20 mg/kg of indomethacin in 1 ml of distilled water was administered orally to each rat using gavage to all groups, except Group A which served as the negative control group.

Study design

Group A (negative control group) consisted of rats given 1 ml of normal saline and sacrificed after 6 h. Group B (positive control group) were rats given 20 mg/kg of indomethacin (n = 4) and sacrificed after 6 h. Group C consists of four rats given 20 mg/kg of indomethacin (n = 4) and treated with aqueous extract of A. sativum (100 mg/kg) body weight, twice daily for 8 days and thereafter sacrificed (n = 4). Group D consisted of four rats given 20 mg/kg of indomethacin (n = 4) and then treated with aqueous extract of A. sativum (200 mg/kg) body weight, twice daily for 8 days and sacrificed (n = 4). Group E consisted of four rats given 20 mg/kg of indomethacin (n = 4) and treated with aqueous extract of A. sativum (300 mg/kg) body weight, twice daily for 8 days and sacrificed (n = 4). Group F consisted of four rats given 20 mg/kg of indomethacin (n = 4) and afterward treated with cimetidine (50 mg/kg) body weight twice daily for 8 days and sacrificed.[15]

At the end of the experiment, the animals were sacrificed following mild anesthesia using chloroform inhalation and the stomach opened along the greater curvature. The gastric lesions induced by indomethacin served as ulcer index (U.I).

Ulcer index

The dissected stomachs were opened along the lesser curvature while the duodenum was also cut opened; the inner surface was rinsed in saline water and examined for ulceration using the Adami et al.[16] method. Magnifying lens was used to measure the length of lesions within 1 mm. U.I was carried out using the modified scoring system of Adami et al. where 0 = no lesion, 1 = hemorrhagic suffusions, 2 = 1–5 small ulcers that are up to 3 mm in length, 3 = many small ulcers more than 5 or 1 ulcer of more than 3 mm, 4 = many ulcers of more than 3 mm, and 5 = perforated ulcers.

The curative ratio (CR) was calculated using the formula below:

CR = (LC − LT/LC) × 100.

Where LC is the length of gastric ulcer in positive group and LT is the length of gastric ulcer in treated group.

Histopathological investigation

Following the dissection and macroscopic examination of the stomach and duodenum, they were fixed in 10% formol saline for 24 h, and tissues were processed in an automatic tissue processor machine (Shandon 2000, Leica, Frankfurt, Germany). While in the processor machine, tissues were dehydrated in different grades of alcohol, cleared in toluene, and impregnated in molten paraffin wax for specified periods. Processed tissues were embedded in fresh molten paraffin wax and allowed to set. Sections were at 3 μ and dried on a hot plate for 15 min and stained with Cole's hematoxylin and 1% aqueous eosin to demonstrate general tissue structure. Stained slides were dehydrated in various ascending grades of alcohol, cleared in xylene, and mounted in Canada balsam.[17] Sections were viewed microscopically using ×10 and ×40 objective lenses.


  Results Top


[Figure 1] shows the effect of indomethacin on the stomach of Wistar rats. [Figure 1]a shows the stomach of Wistar rat that was not exposed to indomethacin. There was no visible ulceration observed. However, there were ulcerations and perforations on the stomach of rats exposed to indomethacin in [Figure 1]b and [Figure 1]c.
Figure 1: Effect of indomethacin on stomach of rats. (a) Stomach without indomethacin (normal macroscopic view. (b) Ulcer as indicated by arrows caused by indomethacin. (c) Perforations caused by indomethacin

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[Figure 2] shows the effect of indomethacin on the duodenum of Wistar rats. Indomethacin produced only visible ulceration on the duodenum as shown in [Figure 2]b while [Figure 2]a shows a normal Wistar rat duodenum not exposed to indomethacin and shows no sign of ulceration or perforation.
Figure 2: Effect of indomethacin on the duodenum of Wistar rats. (a) Normal duodenum rats without indomethacin. (b) Ulcers (indicated by arrows) in the duodenum of Wistar rats given indomethacin

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The effect of treatment with garlic (100–300 mg/kg) on the stomach of Wistar rats are shown on [Figure 3]. In all cases, after 5–8 days of treatment, no visible signs of ulceration or perforation were observed (a and b) in all plates when compared before treatment (c and d). A similar picture was observed in the duodenum following the administration of garlic extract [Figure 4]. In addition, the 200 mg/kg and the 300 mg/kg doses of A. sativum presented with similar results in both the stomach and duodenum.
Figure 3: Effect of garlic treatment (a = 100 mg/kg; b = 300 mg/kg) on the stomach of Wistar rats. (c and d) Ulcers and perforations on the stomach of indomethacin induced rat

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Figure 4: Effect of garlic treatment (a = 100 mg/kg; b = 300 mg/kg) on the duodenum of Wistar rats. (c) Visible ulcers on the duodenum of induced indomethacin rat

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Cimetidine, a standard drug used to treat stomach and duodenal ulcers, was used at a dosage of 50 mg/kg on indomethacin-induced rats. The drug cleared all ulcers and perforations in the stomach [Figure 5] as well as all ulcers in the duodenum [Figure 6].
Figure 5: Effect of cimetidine treatment (a = 50 mg/kg) on the stomach of Wistar rats. (b and c) Visible ulcers and perforations on the stomach of induced indomethacin rat

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Figure 6: Effect of cimetidine treatment (a = 50 mg/kg) on duodenum of Wistar rats. (b) Visible ulcers on the duodenum of induced indomethacin rat

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[Figure 7] shows the effect of indomethacin on sections of the stomach of albino Wistar rats revealing areas of mucosal devitalization, erosion of the villi with ulceration, and gastric pits as represented by the arrows. [Figure 8] shows crater with granulation and inflammatory cells.
Figure 7: Stomach given Indomethacin only showing A, mucosal devitalisation, B erosion, C ulceration and D gastric pit (H and E, ×40)

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Figure 8: Gastric ulcer showing A, crater with B, granulation tissue and C, inflammatory cells (H and E, ×100)

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[Figure 9] shows sections of rat's duodenum given only indomethacin for ulcer revealing devitilized mucosa which composed of necrotic debris, inflammatory cells, and granulation tissue. [Figure 10] shows patchy mucosal ulceration and muscularis mucosa.
Figure 9: Duodenum induced for ulcer showing devitalised mucosa composed of A Necrotic debris, B inflammatory cells and C granulation tissue (H and E, ×40)

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Figure 10: Duodenum given Indomethacin only showing A, patchy mucosal erosion and B, muscularis mucosa (H and E, ×100)

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Sections of stomach and duodenum of albino Wistar rats given 100 mg/kg garlic following the inducement of ulcer by indomethacin showed normal mucosa as shown in [Figure 11] and [Figure 12].
Figure 11: Stomach given 100mg/kg garlic and indomethacin showing A, normal mucosal (H and E, ×40)

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Figure 12: Duodenum given 100mg/kg garlic and indomethacin showing A, fairly normal mucosa (H and E, ×40)

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[Figure 13] and [Figure 14] show sections of rat's stomach and duodenum given 300 mg/kg garlic and indomethacin revealing normal mucosa.
Figure 13: Duodenum given 100mg/kg garlic and indomethacin showing A, fairly normal mucosa (H and E, ×40)

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Figure 14: Duodenum given 300mg/kg Garlic and Indomethacin showing A, fairly normal mucosa (H and E, ×100)

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  Discussion Top


[Figure 1] and [Figure 2] show perforations and ulcerations induced by indomethacin in albino Wistar rats' stomach and duodenum, respectively. This confirms indomethacin as an agent that can induce gastric and duodenal ulcers[1] as Wistar rats not treated with indomethacin have no ulceration and perforation in their stomach and duodenum [Figure 1]a and [Figure 2]a.

A part of the mechanisms by which indomethacin causes ulcer is by increase in the level of oxidative stress as a result of increase in the level of reactive oxygen species (free radicals).[18] These free radicals cause lipid peroxidation which plays a critical role in the development of pathogenesis of acute gastric damage.[19],[20] Aqueous garlic extract is known to exert antioxidant action by scavenging reactive oxygen species, thereby enhancing cellular antioxidant enzymes superoxide dismutase, catalase, and glutathione peroxidase.[21] Garlic represents an important source of antioxidant phytochemicals such as diallyl sulfide, S-allylmercaptocysteine, and ajoene, which is the optimal assurance for neutralizing free radical-mediated inflammation.[22] Aqueous garlic extract inhibits lipid peroxidation and inhibits activation of oxidant-induced transcription factors.[21] These mechanisms may explain the activity of garlic in treating gastric and duodenal ulcers. Indeed, Badr and Al-Mulhim[22] reported that scavenging activity of garlic, the inhibition of lipid peroxidation, its anti-inflammatory action, and antioxidant properties were responsible for its healing and gastroprotective effect of the stomach damaged by indomethacin. These may explain the findings in this study where garlic-treated Wistar rats did not show any sign of macroscopic ulcerations and perforations compared with nongarlic-treated Wistar rats [Figure 3]a,[Figure 3]b,[Figure 3]c,[Figure 3]d. Similar findings were observed in the duodenum of adult Wistar rats treated with garlic after indomethacin was used to induce ulcer [Figure 4]a,[Figure 4]b,[Figure 4]c.

Following treatment with 100 mg/kg garlic, section shows normal mucosa in both the stomach and duodenum [Figure 11] and [Figure 12]. This confirms the macroscopic healing observed in [Figure 3]a and [Figure 4]a. Garlic has been reported to have therapeutic and preventive activities against peptic ulcers.[23] A similar picture was seen in the duodenum, albeit a fairly normal mucosa. This may indicate that garlic is more effective in healing gastric than duodenal ulcers. Therefore, microscopic examination is better index of healing than macroscopic examination.


  Conclusion Top


A. sativum exhibited better healing activities at 100 mg/kg dosage than the 300 mg/kg while cimetidine produced fairly normal mucosa in the stomach and mild mucosa sloughing in the duodenum of the Wistar rats. The 200 mg/kg and the 300 mg/kg doses of A. sativum presented with similar results in the stomach and duodenum. Suffice to say that garlic extract is better than cimetidine in the treatment of gastric and duodenal ulcer. Further studies into the antiulcer activity of A. sativum are advocated.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

 
  References Top

1.
Borra SK, Lagisetty RK, Mallela GR. Anti-ulcer effect of Aloe vera in non-steroidal anti-inflammatory drug induced peptic ulcers in rats. Afr J Pharmacol 2011;5:1867-71.  Back to cited text no. 1
    
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Macpherson LJ, Geierstanger BH, Viswanath V, Bandell M, Eid SR, Hwang S, et al. The pungency of garlic: Activation of TRPA1 and TRPV1 in response to allicin. Curr Biol 2005;15:929-34.  Back to cited text no. 13
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Oguwike FN, Offor CC, Nwadighoha AN, Ebede SO. Evaluation of efficacy of cabbage juice (Brassica oleracea Linne) as potential antiulcer aggent and its effect on the haemostatic mechanism of male albino wistar rats. J Dent Med Sci 2014;13:92-7.  Back to cited text no. 14
    
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Abdel AM, Abdulsalam MA, Ibrahim ND. Effect of aqueous extract of glove (Syzygium aromaticum) on normal gastric secretions and indomethacin induced gastric ulcer in rats. Sci J Al Azhar Med Fac 2006;27:2217-25.  Back to cited text no. 15
    
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Das D, Bandyopadhyay D, Bhattacharjee M, Banerjee RK. Hydroxyl radical is the major causative factor in stress-induced gastric ulceration. Free Radic Biol Med 1997;23:8-18.  Back to cited text no. 20
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Sener G, Sehirli O, Ipçi Y, Ercan F, Sirvanci S, Gedik N, et al. Aqueous garlic extract alleviates ischaemia-reperfusion-induced oxidative hepatic injury in rats. J Pharm Pharmacol 2005;57:145-50.  Back to cited text no. 21
    
22.
Badr GM, Al-Mulhim JA. The protective effect of aged garlic extract on nonsteroidal anti-inflammatory drug-induced gastric inflammations in male albino rats. Evid Based Complement Alternat Med 2014;2014:759642.  Back to cited text no. 22
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Sumiyoshi H. New pharmacological activities of garlic and its constituents. Nihon Yakurigaku Zasshi 1997;110 Suppl 1:93P-7P.  Back to cited text no. 23
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    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7], [Figure 8], [Figure 9], [Figure 10], [Figure 11], [Figure 12], [Figure 13], [Figure 14]



 

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