The rats were dissected and their stomachs were cut open along the greater curvature. The gastric lumen was rinsed with normal saline and examined. The ulcer scoring was carried out using a method described in a previous study. Animals were pre-treated for a period of seven days as described earlier and the rats were fasted for 24 h into the eighth day to ensure complete emptying of the stomach and water was permitted ad libitum.

After anesthetic, the abdomen was opened and pylorus ligation was carried out without causing any damage to its blood supply.

Then, the stomach was replaced carefully and the abdominal wall was closed in two layers with interrupted sutures followed with a moist swab of normal saline.

After 4 h, each stomach was dissected out and cut open along the greater curvature [ 26 , 27 ]. The ulcer index UI was determined using the scoring scaling of Minano et al. After dissection and removal of the stomach, gastric juice was collected.

Than it was centrifuged for 5 min at × g, while the supernatant was separated and used to analyze for pH, gastric juice volume, and total acidity. Total acid was estimated by titrating against 0.

Gastric mucus content was determined using the method described by Corne et al. The stomach was removed, opened along the lesser curvature, and rinsed with cold saline. The levels of superoxide dismutase SOD , glutathione reductase GR , glutathione peroxidase GP , catalase CAT , malondialdehyde MDA and nitrite were determined using a commercially available ELISA kit Cayman, USA [ 30 ].

All the grouped data were presented as mean ± standard error of the mean SEM. The percentage of inhibition was Anti-ulcer effects of virgin coconut oil VCO on cold-stress-induced rats.

Comparatively, the percentage of inhibition for the omeprazole-treated group was As reported above, similar trends were observed for the piroxicam-induced ulcer model. The highest UI 3.

Anti-ulcer effects of virgin coconut oil VCO on piroxicam-induced rats. In the pylorus experimental model, VCO evoked gastric acid secretion in rats. The gastric juice induced by VCO showed significant reduction of total acidity and ulcer scoring. On the other hand, the total acid, for the negative control reported as The gastric mucus content of pylorus ligation negative control rats was 4.

The pH for the negative control group was found to be 2. The UI for the negative control group was 4. The percentage of inhibition noted for the positive control group versus the pylorus-ligated group was Anti-ulcer effects of virgin coconut oil VCO on pylorus ligated model.

negative control. The level of PGE 2 content in the negative control was Anti-ulcer effect of virgin coconut oil VCO on prostaglandin E 2 PGE 2 synthesis. Different factors are involved in the pathogenesis of peptic ulcer in human beings such as chronic use of NSAIDs, stress, H.

pylori infection, alcohol consumption, smoking and inappropriate dietary lifestyle. Although various kinds of medication are available to treat peptic ulcer disease such as H-2 receptor antagonist, proton pump inhibitors PPIs , antacids and anti-muscarinics [ 32 ], most of them cause side effects to patients, yet do not providing a complete recovery [ 33 ].

Earlier studies revealed the effectiveness of VCO in treating ulcer in animal models [ 13 , 19 ]. In the last decade, the exploitation of VCO for their fatty acid and vitamin E composition followed by antioxidant properties and their effects on various ailments have been reported by researchers [ 15 , 16 ].

There has been growing interest and attention on VCO and their potential effects on humans include anti-microbial, anti-viral, anti-diabetic, hypocholesterolemic [ 16 ] and anti-ulcer effect. To the best of our knowledge, no detailed study has been conducted on different ulcer models that elucidated the possible mechanism.

Therefore, the present study was designed to further evaluate the antiulcer activity of VCO compared to a standard drug omeprazole. For this purpose, the effects of VCO were evaluated using different ulcer models induced by cold restraint stress, ethanol, piroxicam NSAIDs , ethanol and pylorus ligation.

Moreover, the effects of VCO on the gastric secretions from a pylorus-ligated model, level of PGE 2 secretion from a piroxicam model and antioxidant assays from an ethanol-induced model were also studied.

Omeprazole is an extensively used anti-ulcer drug belonging to proton pump inhibitors PPIs , to treat peptic ulcer disease caused by stress, non-steroidal anti-inflammatory drugs and by H.

pylori infections [ 34 ]. However, another study revealed that omeprazole acts as a potent antioxidant to scavenge the oxygen free radical and prevents oxidative damage by increasing lipid peroxidation and protein oxidation, in an experiment conducted using three different ulcer models, stress, indomethacin-induced and pylorus ligation-induced models [ 35 ].

Thus, in the present investigation omeprazole was chosen as the standard drug for the positive control treated group. Stress-induced ulcer may be caused by histamine secretion by the parietal cells. In addition, presence of reactive oxygen species ROS due to stress also may provoke damage to he stomach leading to ulceration.

Apart from that, acid and pepsin-associated factors also play a role in the pathogenesis of stress-induced ulcer. Increase in generation of ROS during stress will lead to oxidative damage and cause injury to the mucosal layer. Stress-induced ulcer also leads to a decrease in mucus production.

This finding indicates that VCO may enhance mucus secretion and also play a role in suppressing formation of ROS. As VCO has been reported to contain high anti-oxidant properties followed by flavonoid and other fatty acid components, VCO might contribute to the opposing effect [ 8 ].

From the present investigation, the increase in the levels of GSH and nitrite corresponding to the reduction in MDA, CAT, SOD and GP shown by VCO suggests that there is a strong correlation of its antiulcer activity with the free radical scavenging activity, similar to omeprazole [ 35 ].

Several previous studies have reported on the pathogenesis of ethanol in ulcer induction. It has been proven that exposure to ethanol leads to gastric lesions and mucosal damage [ 36 ]. Similarly, another study mentioned that the occurrence of cellular damage caused by ethanol exposure is dose dependent.

The higher the dose of ethanol, the greater is the damage to the mucosal layer [ 37 ]. Moreover, the involvement of oxygen-derived free radicals has been associated with ethanol-induced ulceration.

Ethanol-induced ulcers are also strongly related to the generation of leukotriene C4 and mast cell secretory products, which also alter the mucosal permeability, reduce gastric mucus and cause severe damage to the gastric mucosal layer [ 38 ].

Therefore, the above information suggests that an agent with high antioxidant properties such as VCO can act as a protectant for the mucosal layer by protecting against necrotizing agents such as ethanol.

This indicates that VCO displays a defensive characteristic against ethanol. As similar trend was observed in cold restraint stress-induced ulcer.

The increase in the levels of GSH and nitrite corresponding to the reduction in MDA, CAT, SOD and GP shown by VCO may also be associated with its defensive action in the ethanol-induced model.

Piroxicam is a known anti-inflammatory drug widely prescribed for patients but it produces side effects and induces ulcer. NSAIDs act via inhibition of cyclooxygenase COX -1 and COX-2 enzymes, which leads to accumulation of intracellular arachidonic acid that inhibits PG synthesis [ 7 ].

Alteration in the PG level promotes acid secretion in the mucosa which disturbs the gastric equilibrium, increases the neutrophil infiltration, induces TNF-α expression and disrupts the balance between nitric oxide NO and other free radical expression [ 39 ].

As PG plays an important role in preventing mucosal injury and shows a protective role via enhancing bicarbonate and mucus production, it is important to prevent the suppression of PG. These results suggest the possibilities of PG and mucus involvement in the anti-ulcer activity of VCO.

Pylorus ligation is a procedure used in enhancing the secretion of gastric acid and breakdown of the gastric mucosal barrier [ 40 ]. This indicates that VCO possesses a protective role in inhibiting ulceration.

Again, the enhanced production of PG as well as up-regulation and down-regulation of antioxidants observed might also be responsible for their anti-ulcerogenic effect.

A previous study reported the presence of phenols, flavonoids, and vitamin E in VCO, which highlights its high antioxidant properties [ 9 ]. Furthermore, presence of some fatty acid components such as lauric acid might also contribute to the effective role of VCO in preventing ulceration.

In conclusion, virgin coconut oil shows potential gastro-protective activity among different kinds of ulcer models. As pathogenesis of peptic ulcer disease is associated with various factors, VCO can be considered as a potential therapy to be used for treating and preventing this ailment.

Current issue Archive Manuscripts accepted About the Journal Editorial office Editorial board Abstracting and indexing Subscription Contact Ethical standards and procedures Most read articles Instructions for authors Article Processing Charge APC Regulations of paying article processing charge APC.

Manuscripts accepted. About the Journal Editorial office Editorial board Abstracting and indexing Subscription Contact Ethical standards and procedures Most read articles. Instructions for authors Article Processing Charge APC Regulations of paying article processing charge APC.

Current issue. Study of the mechanism of anti-ulcer effects of virgin coconut oil on gastric ulcer-induced rat model. Jie Meng 1.

Taoping Chen 1. Yu Zhao 2. Sucai Lu 1. Huiling Yu 1. Ying Chang 1. Dalei Chen 1. Department of Orthopedic, Affiliated Hospital of Hebei University, Baoding, China. Introduction: This study aims to evaluate the gastro-protective effects of virgin coconut oil VCO on different ulcer models as compared to the standard drug omeprazole.

Animals were pre-treated for 7 days and ulcers were induced with cold restraint stress, piroxicam, ethanol and pylorus ligation. On day eight, animals were sacrificed and ulcer scores were determined based on macroscopic evaluation.

The gastric volume, pH, total acidity and mucus content were measured in the pylorus-ligated model. The levels of antioxidants were determined from the gastric tissue homogenates.

Results: Virgin coconut oil significantly p Conclusions: Virgin coconut oil shows a possible association with antioxidant properties to control the regulation of prostaglandin synthesis and protect against reactive oxygen species damage. Introduction Peptic ulcer is a multifactorial and chronic disease [ 1 ].

Material and methods Preparation of virgin coconut oil Low temperature technique was applied in the preparation of VCO. Drugs and chemicals All chemicals and solvents were of analytical grade and included omeprazole Sigma Aldrich, St.

Animal preparation A total of 72 Wistar rats 40 days old , weighing — g, were obtained from our animal feeding center. Anti-ulcer and cytoprotective studies The experimental rats were divided into three groups consisting of 6 rats per group.

Gastric ulcer induction Cold restraint stress-induced gastric ulceration On day 8, the rats were paralyzed by strapping the fore and hind limbs on a flat wooden plank, and transferred into the refrigerator 4—6°C. Ethanol-induced gastric ulceration The experimental rats of each group were pre-treated as explained previously for 7 days and fasted for 24 h into the eighth day.

Piroxicam-induced ulcer model Another set of rats were pre-treated for 7 days as described earlier. Pylorus-ligated rats Animals were pre-treated for a period of seven days as described earlier and the rats were fasted for 24 h into the eighth day to ensure complete emptying of the stomach and water was permitted ad libitum.

Measurement of gastric juice volume, total acidity, pH and gastric mucus content After dissection and removal of the stomach, gastric juice was collected. Analysis Antioxidant enzyme assay The levels of superoxide dismutase SOD , glutathione reductase GR , glutathione peroxidase GP , catalase CAT , malondialdehyde MDA and nitrite were determined using a commercially available ELISA kit Cayman, USA [ 30 ].

Statistical analysis All the grouped data were presented as mean ± standard error of the mean SEM. Table I Anti-ulcer effects of virgin coconut oil VCO on cold-stress-induced rats. Table II Anti-ulcer effects of virgin coconut oil VCO on ethanol-induced rats.

Effects of virgin coconut oil on piroxicam-induced ulcer As reported above, similar trends were observed for the piroxicam-induced ulcer model. Table III Anti-ulcer effects of virgin coconut oil VCO on piroxicam-induced rats.

Effects of virgin coconut oil on pylorus ligation-induced ulcer model In the pylorus experimental model, VCO evoked gastric acid secretion in rats.

Table IV Anti-ulcer effects of virgin coconut oil VCO on pylorus ligated model. Table V Anti-ulcer effects of virgin coconut oil VCO on antioxidant assays. Effects of virgin coconut oil on mucosal prostaglandin E 2 content The level of PGE 2 content in the negative control was Table VI Anti-ulcer effect of virgin coconut oil VCO on prostaglandin E 2 PGE 2 synthesis.

Discussion Different factors are involved in the pathogenesis of peptic ulcer in human beings such as chronic use of NSAIDs, stress, H. Conflict of interest The authors declare no conflict of interest.

Arrieta J , Benitez J , Flores E , Castillo C , Navarrete A. Purification of gastroprotective triterpenoids from the stem bark of Amphipterygium adstringens: role of prostaglandins, sulfhydryls, nitric oxide and capsaicin-sensitive neurons Planta Medica.

Google Scholar. Mustafa M , Menon J , Muiandy RK , Fredie R , Sein MM , Fariz A. Risk factors, diagnosis, and management of peptic ulcer disease J Dental Med Sci.

Sung J , Kuipers E , El-serag H. Systematic review: the global incidence and prevalence of peptic ulcer disease Aliment Pharm Ther.

Barbastefano V , Cola M , Luiz-Ferreira A , et al. Vernonia polyanthes as a new source of antiulcer drugs Fitoterapia. Ode OJ , Asuzu OV. Investigation of cassia singuena leaf extract for antiulcer effects using ethanol-induced gastric ulcer model in rats Int J Plant Animal Environ Sci. Sabiu S , Garuba T , Sunmonu T , et al.

Despite the unknown mechanisms of ethanol-mediated gastric ulcers, still, multiple pieces of evidence show that pro-inflammatory cytokines, oxidative damage, and cellular apoptosis play a vital role in its ethanol-induced progression Al Batran et al.

Various additional factors, such as the environment cigarettes, alcohol, and infectious agents , chronic use of pain relievers such as nonsteroidal anti-inflammatory medicines, and incidents of stress, are among the leading causes of stomach ulcers Tabiri et al.

Changes in the stomach mucosal barrier, blood flow, and degenerative gastric secretion are all contributors to gastric ulcers in terms of major pathophysiology Poonam et al.

Due to disturbance in the mucosal blood flow, angiogenesis, and reduction of cell proliferation, ulcer healing in cigarette smokers is delayed Zhang et al.

Helicobacter pylori H. pylori is a Gram-negative bacterium that can cause chronic gastritis, peptic ulcers, gastric adenocarcinoma, and mucosa-associated lymphoid tissue lymphoma in humans. The infection caused by this pathogen estimates about half of the world's population Zamani et al.

However, infection rates vary by region, with developing countries having a higher frequency than developed countries. All stimulations of stomach acid production lead to the proton pump which is the common and final pathway. Ethanol causes changes in the cytokine equilibrium in the stomach mucosa, which causes inflammation Almasaudi et al.

Antioxidants seemed to have a protective role in gastric ulcers and carcinomas, whereas any imbalance in the activity of these oxidative stress enzymes usually results in improper free radical disposal, which results in the ulceration of gastric tissues Tandon et al. Triple-based eradication therapy, which comprises a proton pump inhibitor PPI and two antimicrobial drugs, amoxicillin and clarithromycin, is thought to be the most effective treatment for H.

pylori -induced ulcer eradication Tulassay et al. Currently, commercially available worldwide PPIs are omeprazole, lansoprazole, and pantoprazole. Proton pump inhibitors have been the treatment of choice for stomach acid hypersecretion Richardson et al. Joint discomfort, irregular heartbeat, hemopoietic changes, gynecomastia, impotence, and systemic alkalosis are all common side effects of these medicines.

Nowadays, more research is being conducted on the utilization of natural products in the development of medications with fewer adverse effects Mahmoud and Ghffar, The ethanol-induced gastric ulcer rodent model is considered a well-established model that reflects several features of human gastric injury and thus provides a means for testing unexplored test samples with anti-ulcer potential Liu et al.

Brucine Figure 1 , one of the principal bioactive constituents isolated from the seeds of Strychnos nux-vomica L. Loganiaceae , is a weak indole alkaloid.

Maqianzi, also known as Nux-vomica, has a bitter flavor Lu et al. Wide pharmacological activities of brucine have been reported by many basic and clinical researchers such as anti-tumor Qin et al. To date, to the best of our knowledge, no detailed study has been conducted that highlights the possible therapeutic potential of brucine in gastric ulcers.

The current study has been conducted to evaluate the brucine anti-ulcer effect using molecular, in silico , in vitro , and in vivo techniques. All standard chemicals used were purchased from verified sources.

Brucine was purchased from Sigma CO, Merck. Normal saline, absolute ethanol, and chloroform were acquired from Sigma-Aldrich St. Louis, MO, United States. Omeprazole and metronidazole were purchased from Barrett Hodgson and Sanofi Aventis, Pakistan. Abcam, United Kingdom, provided secondary antibodies.

Elabscience, China, provided the rat NFB ELISA kit catalog no. E-EL-R , rat TNF-α ELISA kit catalog no. E-EL-ROO19 , and rat PGE2 ELISA kit catalog no. All of the chemicals used in this experiment were of analytical quality. Sprague—Dawley rats weighing — g of either gender, purchased from the Riphah Institute of Pharmaceutical Sciences RIPS in Islamabad, were subjected to conduct this research work.

The rats were kept in standard cages with a constant temperature of 22°C. All of the animals were fed a regular meal and had unlimited access to water. The experiments were performed by following the guidelines and principles of the Institute of Laboratory Animal Resources, Commission on Life Sciences University, National Research Council The study was approved by the Research and Ethics Committee of RIPS Ref.

The three-dimensional structure of a typical medication was obtained using a Biovia Discovery Studio Visualizer DSV. The standard drug omeprazole was used.

In gastric ulcer pathophysiology, human protein targets 3D structure involved were selected and acquired from an online protein data bank, Research Collaboratory for Structural Bioinformatics RCSB PDB.

The ligand and water molecules were removed from DSV, H-atoms polar were inserted, and the file was saved in PDB format. For molecular docking, the AutoDock tool By the disc diffusion method, the antibacterial activity of the selected natural compound against H.

pylori was evaluated. From the gastric biopsy of a patient with gastric ulcer, three strains of H. pylori were received under consent at the care endoscopy clinics and laboratories Rawalpindi, Pakistan. Biopsies to be analyzed were placed in a modified Campy-Thio medium.

Plates were kept at 37°C in a microaerophilic environment. Identification of isolates was carried out by morphology and using a urease test kit. Isolates were kept at 80°C in sterile McCartney bottles containing 0. Frozen clinical isolates were injected onto Mueller—Hinton agar plates for subsequent inoculation.

Brucine with various concentrations of 0. Following an incubation period of 3 to 5 days at 37°C, the zone of inhibition for each disk was measured. All tests were carried out in triplicate, and the antibacterial activity was calculated as the average inhibitory diameter mm.

Metronidazole was used as a positive control Foroumadi et al. All rats were sacrificed by cervical dislocation after 1 h of ethanol administration. The stomachs of the participants were separated and cleaned with normal saline before the lesion index was computed by measuring each lesion in millimeters along its greater curvature.

For each stomach, the sum of the length mm of all lesions was used as the ulcer index UI. The homogenate of stomach tissue was centrifuged for 10—15 min at 3, rpm. As a result of the supernatant, spectrophotometrically at nm, the inorganic phosphate release was determined.

One ATPase activity unit has been described as one µ mole of inorganic phosphorus released by ATP hydrolysis through ATPase at 1 mg per hour of tissue protein. The antioxidant activity of brucine was analyzed in the isolated tissues of the pretreated animals and compared with disease and positive control group tissues.

The stomach tissues were homogenized and centrifuged at 1, rpm for 30 min to collect the supernatant. The obtained supernatant was estimated for glutathione GSH , catalase, glutathione-S-transferase GST , and lipid peroxidation LPO.

The oxidation of GSH and DTNP produced a yellow end product, which was used to determine their levels. Using a GSH microplate reader, the absorbance of 2-nitrothiobenzoic acid was determined at nm.

The synthesis of the CDNB conjugate was used to determine the GST level, and the absorbance at nm was measured. In the presence of catalase, the degradation of H 2 O 2 was measured. A microplate reader was used to measure absorbance at nm.

The end product of LPO, malondialdehyde, was used to determine the level of LPO MDA. On coated slides, in percent pure xylene, tissue sections were deparaffinized and then rehydrated in 70 percent ethyl alcohol.

The slides were immersed in hematoxylin for 10 min after being cleaned with distilled water. The slides were immersed in eosin solution for 5—10 min. After the required length of time had passed, the slides were cleaned in water and air—dried. In graded ethyl alcohol, the dried slides were dehydrated 70 percent, 95 percent, and percent.

The slides were cleaned with xylene and mounted with glass coverslips. The slides were photographed using an Olympus light microscope and analyzed by ImageJ, a computer-based application, with particular attention paid to the gastric cell size and shape, inflamed infiltrating cells, and vacuolation.

The TIF images were arranged at the same threshold intensity for all groups and examined in GraphPad Prism. The immunohistochemical examination was carried out, as previously described by Gim et al. The slides were deparaffinized, then treated for antigen retrieval enzymatic technique , and washed with PBS.

For 10 min, 3 percent hydrogen peroxide was used to block the endogenous peroxidase in methanol H 2 O 2. The slides were incubated for a period of time in a solution containing 0.

After being blocked, the slides were treated overnight with murine anti-TNF-α, p-NFkB, and anti-COX-2 antibodies dilution , Santa Cruz Biotechnology. Slides were handled for incubation with the biotinylated secondary antibody dilution , depending on the source of the primary antibody, and the serum was used the next morning after cleaning with 0.

The ABC Elite kit was used to incubate slides in a humidified room for 1 h after secondary antibody application Santa Cruz Biotechnology. The slides were cleaned in 0. A light microscope was used to capture immunohistochemical TIF pictures. By modifying the backdrop of photos according to the threshold intensity and analyzing p-NFκB, COX-2, and TNF-α positive cells at the same threshold intensity for all groups, ImageJ software was utilized to quantitatively detect hyperactivated p-NFκB, COX-2, and TNF-α.

The relative integrated density of the samples compared to saline is used to compute the intensity Ansari et al. Using the Silent Crusher M Heidolph apparatus, the tissues were homogenized at 15, RPM; after centrifugation, the supernatant was collected for 1 h at 1, g.

TNF-α, PGE2, and p-NFκB concentrations were measured using an ELISA microplate reader. Ansari et al. Gastric tissues were homogenized after being lysed in a buffer. A bicinchoninic acid BCA protein test kit was used to assess the protein content. On a 12 percent sodium dodecyl sulfate-polyacrylamide gel, 30 g of protein homogenate was electrophoretically separated and transferred to a polyvinylidene fluoride membrane.

After being cleansed three times with Tris-buffered saline with 0. To see the immunoreactive bands, an enhanced Western blotting substrate kit was used. Densitometry was used to assess the measurement of protein expression by ImageJ software. Irshad et al. The TRIzol technique was used to extract total ribonucleic acid RNA from gastric tissues, as directed by the manufacturer.

Using a reverse transcriptase enzyme mix and a PCR thermocycler, the first strand of cDNA was generated from 1 to 2 g of total RNA. The data are presented as a mean with a standard error of the mean SEM. In this study, brucine binds to a variety of protein receptors with varying affinities.

Supplementary Figures S1—S8 illustrate the 2D depiction of brucine interactions with common medications. Brucine had an E-value of Brucine has an E-value of —2. Table 1. TABLE 1.

The minimum inhibitory concentration MIC and zone of inhibition of three different strains of H. pylori against brucine were measured.

Different concentrations of brucine were used against strain A that are 0. The same concentration of brucine was used for strain B , and the inhibition values are 1.

For strain C , same concentrations of brucine were used, and the inhibitory values are 1. TABLE 2. Antibacterial effect of brucine against three clinical strains of H.

pylori using the disk diffusion method. The stomach mucosa of rats was observed under a microscope Figure 2. TABLE 3. Protective effect of brucine and omeprazole against ethanol-induced gastric ulcer in rats.

FIGURE 2. FIGURE 3. ethanol group. In ethanol-induced ulcer stomach tissues, GST, GSH, and catalase levels were decreased, but LPO levels were increased. FIGURE 4. Effect of brucine and omeprazole against glutathione sulfotransferases GST , reduced glutathione GSH , catalase, and lipid peroxidase LPO in ethanol-induced ulcer rat gastric tissues.

FIGURE 5. Histopathological slides represent effect of brucine and omeprazole in ethanol- induced ulcer rats gastric tissues. Saline group showing normal histological features. Ethanol ulcer group showing marked histopathological deformities by loss of stomach architecture, vacuolization and cloudy swelling.

Brucine and omeprazole treatment groups showing near normal architecture with mild to moderate deformities. Vacuolation, necrotic cells, and disruption of morphological cell boundaries were found in the disease group.

FIGURE 6. Slides represent the effect of brucine and omeprazole against the expressions of cyclooxygenase COX-2 , nuclear factor kappa B p-NFκB , and tumor necrosis factor alpha TNF-α in ethanol-induced ulcer rat gastric tissues, using the immunohistochemical technique.

FIGURE 7. Effects of brucine and omeprazole against cyclooxygenase COX-2 , nuclear factor kappa B p-NFκB , and tumor necrosis factor alpha TNF-α in ethanol-induced ulcer rat gastric tissues, using the immunohistochemical technique.

saline group. FIGURE 8. Effects of brucine and omeprazole against nuclear factor kappa B p-NFκB , prostaglandin PGE 2 , and tumor necrosis factor TNF-α levels in ethanol-induced ulcer rat gastric tissues, using the enzyme-linked immunosorbent assay technique.

FIGURE 9. Bands A and graphical B representation of effects of brucine and omeprazole against phosphorylated nuclear factor kappa B p-NFκB and tumor necrosis factor TNF-α expressions in ethanol-induced ulcer rat gastric tissues, using the Western blot technique.

FIGURE This research work was conducted to explore the protective actions of brucine against ethanol-induced gastric ulcers in Sprague—Dawley rats. Ethanol is known for its notorious gastric ulceration by directly enhancing the disruption of mucosal cellular membranes, dehydration, and cytotoxicities, followed by the propagation of inflammatory cytokines, oxidative damage, and apoptosis Park et al.

The currently used model of ethanol-induced gastric ulcer is a well-established rodent model commonly implicated for preclinical evaluation of new drug molecules having anti-ulcer potential since ethanol has been regarded as a leading cause of gastric injuries in humans Augusto et al.

Drug discovery and development is incomplete without the evaluation of the structure of a compound. For this purpose, computational studies were carried out where the ligand was docked with its respective target using compound and protein databases Nayarisseri, This study displayed results of all ligands compared with standard drugs, obtained from the PubChem database and RCSB.

In the present study, the ethanol-induced gastric ulcer model was used for in vivo experimentation Li et al. In an in vivo study, pretreatment of rats with brucine at both doses significantly reduced the ulcer index relative to the disease group.

In comparison to the usual treatment, ulcerated animals pretreated with brucine demonstrated a greater reduction in the ulcer index as compared to omeprazole Abebaw et al. In human and experimental animals, oxygen-derived free radicals have been implicated in the etiology of a wide range of clinical diseases and stomach injuries, resulting in gastrointestinal ulcers Yeo et al.

In the in vitro conformational analysis, H. pylori is the main risk factor for gastric ulcer disease Graham, Brucine possesses antibacterial activity as it inhibits H.

Moreover, VCO is an edible food which can be included in our diet and taken directly without prescription from a physician, without causing any side effects. Although studies have reported on the effects of VCO on peptic ulcer [ 13 ], there has been no study yet to elucidate the mechanism involved in inhibiting ulceration.

The present study aims to reveal the possible mechanism involved in the action of VCO in ulcerative models. Low temperature technique was applied in the preparation of VCO. The solid endosperm of matured fresh coconut was crushed and made into a viscous slurry [ 16 ].

It was later mechanically squeezed through a cheese cloth to collect the coconut milk. The milk was kept in the refrigerator and frozen for 48 h, then heated at °C using a thermostat oven. After the heating process, the collected oil was filtered using a cheese cloth.

All chemicals and solvents were of analytical grade and included omeprazole Sigma Aldrich, St. Louis, MD, USA , ethanol Merck, UK , phenolphthalein Merck, UK , alcian blue Merck, UK , magnesium chloride Merck, UK , sucrose Merck, UK and sodium citrate Merck, UK.

A total of 72 Wistar rats 40 days old , weighing — g, were obtained from our animal feeding center. All studies carried out were approved by the animal care and ethical committee following the employed protocols ACUC No: HDFY-LL The animals were kept at room temperature 25—28°C for a minimum of seven days for acclimatization to room conditions prior to the experimental procedure.

The experimental rats were divided into three groups consisting of 6 rats per group. The grouping was the same for each ulcer induction model. On day 8, the rats were paralyzed by strapping the fore and hind limbs on a flat wooden plank, and transferred into the refrigerator 4—6°C.

After two hours, the rats were sacrificed by cervical dislocation. The stomach was incised along the greater curvature and the lumen was rinsed with normal saline.

The ulcer scoring was carried out based on the method described by Minano et al. The acquired supernatant was used to study activity using an antioxidant enzyme assay. The experimental rats of each group were pre-treated as explained previously for 7 days and fasted for 24 h into the eighth day.

After 1 h of ulcer induction, the animals were sacrificed by cervical dislocation. The stomach was dissected out for the macroscopic examination of ulcers. The scoring for ulcer was determined using a method described in a previous study.

Another set of rats were pre-treated for 7 days as described earlier. The animals were then fasted for 24 h into the eighth day. After 6 h of piroxicam induction, the animals were sacrificed by cervical dislocation. The rats were dissected and their stomachs were cut open along the greater curvature.

The gastric lumen was rinsed with normal saline and examined. The ulcer scoring was carried out using a method described in a previous study. Animals were pre-treated for a period of seven days as described earlier and the rats were fasted for 24 h into the eighth day to ensure complete emptying of the stomach and water was permitted ad libitum.

After anesthetic, the abdomen was opened and pylorus ligation was carried out without causing any damage to its blood supply. Then, the stomach was replaced carefully and the abdominal wall was closed in two layers with interrupted sutures followed with a moist swab of normal saline.

After 4 h, each stomach was dissected out and cut open along the greater curvature [ 26 , 27 ]. The ulcer index UI was determined using the scoring scaling of Minano et al. After dissection and removal of the stomach, gastric juice was collected.

Than it was centrifuged for 5 min at × g, while the supernatant was separated and used to analyze for pH, gastric juice volume, and total acidity. Total acid was estimated by titrating against 0. Gastric mucus content was determined using the method described by Corne et al.

The stomach was removed, opened along the lesser curvature, and rinsed with cold saline. The levels of superoxide dismutase SOD , glutathione reductase GR , glutathione peroxidase GP , catalase CAT , malondialdehyde MDA and nitrite were determined using a commercially available ELISA kit Cayman, USA [ 30 ].

All the grouped data were presented as mean ± standard error of the mean SEM. The percentage of inhibition was Anti-ulcer effects of virgin coconut oil VCO on cold-stress-induced rats.

Comparatively, the percentage of inhibition for the omeprazole-treated group was As reported above, similar trends were observed for the piroxicam-induced ulcer model.

The highest UI 3. Anti-ulcer effects of virgin coconut oil VCO on piroxicam-induced rats. In the pylorus experimental model, VCO evoked gastric acid secretion in rats. The gastric juice induced by VCO showed significant reduction of total acidity and ulcer scoring.

On the other hand, the total acid, for the negative control reported as The gastric mucus content of pylorus ligation negative control rats was 4. The pH for the negative control group was found to be 2. The UI for the negative control group was 4. The percentage of inhibition noted for the positive control group versus the pylorus-ligated group was Anti-ulcer effects of virgin coconut oil VCO on pylorus ligated model.

negative control. The level of PGE 2 content in the negative control was Anti-ulcer effect of virgin coconut oil VCO on prostaglandin E 2 PGE 2 synthesis. Different factors are involved in the pathogenesis of peptic ulcer in human beings such as chronic use of NSAIDs, stress, H.

pylori infection, alcohol consumption, smoking and inappropriate dietary lifestyle. Although various kinds of medication are available to treat peptic ulcer disease such as H-2 receptor antagonist, proton pump inhibitors PPIs , antacids and anti-muscarinics [ 32 ], most of them cause side effects to patients, yet do not providing a complete recovery [ 33 ].

Earlier studies revealed the effectiveness of VCO in treating ulcer in animal models [ 13 , 19 ]. In the last decade, the exploitation of VCO for their fatty acid and vitamin E composition followed by antioxidant properties and their effects on various ailments have been reported by researchers [ 15 , 16 ].

There has been growing interest and attention on VCO and their potential effects on humans include anti-microbial, anti-viral, anti-diabetic, hypocholesterolemic [ 16 ] and anti-ulcer effect.

To the best of our knowledge, no detailed study has been conducted on different ulcer models that elucidated the possible mechanism. Therefore, the present study was designed to further evaluate the antiulcer activity of VCO compared to a standard drug omeprazole. For this purpose, the effects of VCO were evaluated using different ulcer models induced by cold restraint stress, ethanol, piroxicam NSAIDs , ethanol and pylorus ligation.

Moreover, the effects of VCO on the gastric secretions from a pylorus-ligated model, level of PGE 2 secretion from a piroxicam model and antioxidant assays from an ethanol-induced model were also studied.

Omeprazole is an extensively used anti-ulcer drug belonging to proton pump inhibitors PPIs , to treat peptic ulcer disease caused by stress, non-steroidal anti-inflammatory drugs and by H.

pylori infections [ 34 ]. However, another study revealed that omeprazole acts as a potent antioxidant to scavenge the oxygen free radical and prevents oxidative damage by increasing lipid peroxidation and protein oxidation, in an experiment conducted using three different ulcer models, stress, indomethacin-induced and pylorus ligation-induced models [ 35 ].

Thus, in the present investigation omeprazole was chosen as the standard drug for the positive control treated group. Stress-induced ulcer may be caused by histamine secretion by the parietal cells. In addition, presence of reactive oxygen species ROS due to stress also may provoke damage to he stomach leading to ulceration.

Apart from that, acid and pepsin-associated factors also play a role in the pathogenesis of stress-induced ulcer.

Increase in generation of ROS during stress will lead to oxidative damage and cause injury to the mucosal layer. Stress-induced ulcer also leads to a decrease in mucus production. This finding indicates that VCO may enhance mucus secretion and also play a role in suppressing formation of ROS.

As VCO has been reported to contain high anti-oxidant properties followed by flavonoid and other fatty acid components, VCO might contribute to the opposing effect [ 8 ]. From the present investigation, the increase in the levels of GSH and nitrite corresponding to the reduction in MDA, CAT, SOD and GP shown by VCO suggests that there is a strong correlation of its antiulcer activity with the free radical scavenging activity, similar to omeprazole [ 35 ].

Several previous studies have reported on the pathogenesis of ethanol in ulcer induction. It has been proven that exposure to ethanol leads to gastric lesions and mucosal damage [ 36 ].

Similarly, another study mentioned that the occurrence of cellular damage caused by ethanol exposure is dose dependent. The higher the dose of ethanol, the greater is the damage to the mucosal layer [ 37 ].

Moreover, the involvement of oxygen-derived free radicals has been associated with ethanol-induced ulceration. Ethanol-induced ulcers are also strongly related to the generation of leukotriene C4 and mast cell secretory products, which also alter the mucosal permeability, reduce gastric mucus and cause severe damage to the gastric mucosal layer [ 38 ].

Therefore, the above information suggests that an agent with high antioxidant properties such as VCO can act as a protectant for the mucosal layer by protecting against necrotizing agents such as ethanol.

This indicates that VCO displays a defensive characteristic against ethanol. As similar trend was observed in cold restraint stress-induced ulcer. The increase in the levels of GSH and nitrite corresponding to the reduction in MDA, CAT, SOD and GP shown by VCO may also be associated with its defensive action in the ethanol-induced model.

Piroxicam is a known anti-inflammatory drug widely prescribed for patients but it produces side effects and induces ulcer. NSAIDs act via inhibition of cyclooxygenase COX -1 and COX-2 enzymes, which leads to accumulation of intracellular arachidonic acid that inhibits PG synthesis [ 7 ]. Alteration in the PG level promotes acid secretion in the mucosa which disturbs the gastric equilibrium, increases the neutrophil infiltration, induces TNF-α expression and disrupts the balance between nitric oxide NO and other free radical expression [ 39 ].

As PG plays an important role in preventing mucosal injury and shows a protective role via enhancing bicarbonate and mucus production, it is important to prevent the suppression of PG. These results suggest the possibilities of PG and mucus involvement in the anti-ulcer activity of VCO.

Pylorus ligation is a procedure used in enhancing the secretion of gastric acid and breakdown of the gastric mucosal barrier [ 40 ]. This indicates that VCO possesses a protective role in inhibiting ulceration. Again, the enhanced production of PG as well as up-regulation and down-regulation of antioxidants observed might also be responsible for their anti-ulcerogenic effect.

A previous study reported the presence of phenols, flavonoids, and vitamin E in VCO, which highlights its high antioxidant properties [ 9 ]. Furthermore, presence of some fatty acid components such as lauric acid might also contribute to the effective role of VCO in preventing ulceration.

In conclusion, virgin coconut oil shows potential gastro-protective activity among different kinds of ulcer models. As pathogenesis of peptic ulcer disease is associated with various factors, VCO can be considered as a potential therapy to be used for treating and preventing this ailment.

Current issue Archive Manuscripts accepted About the Journal Editorial office Editorial board Abstracting and indexing Subscription Contact Ethical standards and procedures Most read articles Instructions for authors Article Processing Charge APC Regulations of paying article processing charge APC.

Manuscripts accepted. About the Journal Editorial office Editorial board Abstracting and indexing Subscription Contact Ethical standards and procedures Most read articles.

Instructions for authors Article Processing Charge APC Regulations of paying article processing charge APC. Current issue. Study of the mechanism of anti-ulcer effects of virgin coconut oil on gastric ulcer-induced rat model. Jie Meng 1. Taoping Chen 1. Yu Zhao 2.

Sucai Lu 1. Huiling Yu 1. Ying Chang 1. Dalei Chen 1. Department of Orthopedic, Affiliated Hospital of Hebei University, Baoding, China. Introduction: This study aims to evaluate the gastro-protective effects of virgin coconut oil VCO on different ulcer models as compared to the standard drug omeprazole.

Animals were pre-treated for 7 days and ulcers were induced with cold restraint stress, piroxicam, ethanol and pylorus ligation. On day eight, animals were sacrificed and ulcer scores were determined based on macroscopic evaluation.

The gastric volume, pH, total acidity and mucus content were measured in the pylorus-ligated model. The levels of antioxidants were determined from the gastric tissue homogenates.

Results: Virgin coconut oil significantly p Conclusions: Virgin coconut oil shows a possible association with antioxidant properties to control the regulation of prostaglandin synthesis and protect against reactive oxygen species damage. Introduction Peptic ulcer is a multifactorial and chronic disease [ 1 ].

Material and methods Preparation of virgin coconut oil Low temperature technique was applied in the preparation of VCO. Drugs and chemicals All chemicals and solvents were of analytical grade and included omeprazole Sigma Aldrich, St. Animal preparation A total of 72 Wistar rats 40 days old , weighing — g, were obtained from our animal feeding center.

Anti-ulcer and cytoprotective studies The experimental rats were divided into three groups consisting of 6 rats per group.

Gastric ulcer induction Cold restraint stress-induced gastric ulceration On day 8, the rats were paralyzed by strapping the fore and hind limbs on a flat wooden plank, and transferred into the refrigerator 4—6°C.

Ethanol-induced gastric ulceration The experimental rats of each group were pre-treated as explained previously for 7 days and fasted for 24 h into the eighth day.

Piroxicam-induced ulcer model Another set of rats were pre-treated for 7 days as described earlier. GERD is caused by excessive hydrochloric acid that tends to back up, or reflux, into the lower esophagus.

See Figure 7. Peptic ulcer disease PUD occurs when gastric or duodenal ulcers are caused by the breakdown of GI mucosa by pepsin, in combination with the caustic effects of hydrochloric acid.

PUD is the most harmful disease related to hyperacidity because it can result in bleeding ulcers, a life-threatening condition. Stress-related mucosal damage is another common condition that can occur in hospitalized patients leading to PUD. Thus, many post-operative or critically ill patients receive medication to prevent the formation of a stress ulcer, which is also called prophylaxis.

Links to supplementary videos illustrating heartburn and gastric ulcers:. Heartburn [5]. Gastric ulcer [6]. Assessments: Whenever a nurse administers hyperacidity medications, there are common assessments that should be documented, such as an abdominal assessment and documentation of bowel patterns.

During therapy, the nurse should continue to assess for potential medication interactions and side effects and be aware that vitamin B12 malabsorption may occur whenever stomach acidity levels are altered.

Based on the category of medication, renal and liver function may require monitoring. Implementation: The nurse should read the drug label information and follow the recommendations for administering hyperacidity medications with other medications or the intake of food. Cultural preferences should also be accommodated when safe and feasible because the patient may believe in alternative methods for treating GI discomfort.

A written plan of care with modifications for safe use of medications with these alternative methods may be required. Evaluation: Patients should experience improvement of symptoms within the defined time period; if not, the provider should be notified.

There are four major classes of medications used to treat hyperacidity conditions: antacids, H2-receptor antagonists, proton pump inhibitors, and mucosal protectants.

Each class of medication is further described below. Antacids see Figure 7. There are many OTC medications available for this purpose, such as calcium carbonate, aluminum hydroxide, and magnesium hydroxide.

Calcium carbonate is the prototype discussed as an example. Be sure to read drug label information regarding antacids as you administer them because each type has its own specific side effects. Many antacids also contain simethicone, an antiflatulent used for gas relief.

Simethicone is further described in the medication grid below. Antacids neutralize gastric acidity and elevate the pH of the stomach. Elevated pH also inactivates pepsin, a digestive enzyme.

Calcium carbonate comes in various formations such as a tablet, a chewable tablet, a capsule, or liquid to take by mouth. It is usually taken three or four times a day. Chewable tablets should be chewed thoroughly before being swallowed; do not swallow them whole. The patient should drink a full glass of water after taking either the regular or chewable tablets or capsules.

Some liquid forms of calcium carbonate must be shaken well before use. Do not administer calcium carbonate within hours of other medicines because calcium may decrease the effectiveness of the other medicine.

Calcium carbonate may be contraindicated in patients with preexisting kidney disease because it may cause hypercalcemia. Common side effects of calcium carbonate include constipation and rebound hyperacidity when it is discontinued. Other interventions to prevent hyperacidity can also be recommended, such as smoking cessation and avoiding food and beverages that can cause increased acidity alcohol, high-fat or spicy foods, and caffeine.

A common H2-receptor antagonist is famotidine. It is available OTC and is also often prescribed orally or as an IV injection in the hospital setting. Other H2-receptor antagonists include cimetidine and ranitidine. Cimetidine has a high risk of drug interactions, especially in elderly patients because of its binding to cytochrome P enzymes in the liver, which affects the metabolism of other drugs.

Famotidine see Figure 7. OTC famotidine is also used to treat heartburn or sour stomach. To prevent symptoms, oral famotidine is taken 15 to 60 minutes before eating foods or drinking drinks that may cause heartburn.

Preexisting liver and kidney disease may require dosage adjustment. Famotidine is supported by evidence as safe for use in pediatric patients younger than 1 year old, as well as in geriatric patients. Patients taking the oral suspension should be instructed to shake it vigorously for 5 to 10 seconds prior to each use.

Additionally, smoking interferes with histamine antagonists and should be discouraged. A common proton pump inhibitor PPI is pantoprazole see Figure 7. It may be prescribed in various routes including orally, with an NG tube, or as an IV injection in the hospital setting.

Other PPIs include esomeprazole, lansoprazole, and omeprazole. PPIs are more powerful than antacids and H2-receptor antagonists. Pantoprazole is used to treat damage from gastroesophageal reflux disease GERD in adults and children five years of age and older by allowing the esophagus to heal and prevent further damage.

It is also used to treat conditions where the stomach produces too much acid, such as Zollinger-Ellison syndrome in adults. PPIs may also be given in combination with antibiotics to treat H.

Pylori infections, a common cause of duodenal ulcers. PPIs inhibit the secretion of hydrochloric acid, and the antisecretory effect lasts longer than 24 hours. Packets of delayed-release granules must be mixed with applesauce or apple juice and taken by mouth or given through a feeding tube.

Consult the labeling of concomitantly used drugs to obtain further information about interactions because PPIs can interfere with the liver metabolism of other drugs. IV pantoprazole can potentially exacerbate zinc deficiency, and long-term therapy can cause hypomagnesemia, so the nurse should monitor for these deficiencies.

In addition to the considerations above, instruct patients to call their provider if their condition does not improve or gets worse, especially if bleeding occurs.

Sucralfate locally covers the ulcer site in the GI tract and protects it against further attack by acid, pepsin, and bile salts. It is minimally absorbed by the gastrointestinal tract. Administer sucralfate on an empty stomach, 2 hours after or 1 hour before meals.

Constipation may occur. Sucralfate should be cautiously used with patients with chronic renal failure or those receiving dialysis due to impaired excretion of small amounts of absorbed aluminum that can occur with sucralfate.

In addition to the considerations above, instruct patients to call their provider if their condition does not improve or gets worse. Simethicone is an antiflatulent that is commonly found in other OTC antacids see Figure 7.

It is also safe for use in infants. Gas commonly occurs in the GI tract due to digestive processes and the swallowing of air. Gaseous distension can also occur postoperatively.

Simethicone is used to treat the symptoms of gas such as uncomfortable or painful pressure, fullness, and bloating. Simethicone works by altering the elasticity of the mucous-coated gas bubbles, which cause them to break into smaller bubbles, thus reducing pain and facilitating expulsion.

Simethicone is usually taken four times a day, after meals and at bedtime. For liquid form, shake drops before administering. Patients can be instructed about other measures to assist with gas expulsion such as changing position, ambulation, avoiding the use of straws, and tapering intake of beans and cruciferous vegetables.

Medication grids are intended to assist students to learn key points about each medication.