Al-Razzuqi R. A, Abu-Rageef A. R, Mehasin W. S, Al-Razzuqi T. R. A. Evaluation of Gastroprotective Effect of Vanadyl Sulfate and Lycopene on rat model with Ethanol-Induced Gastric Mucosal Lesions. Biomed Pharmacol J 2018;11(3).

---

Manuscript received on :4-Jun-2018
Manuscript accepted on :13-Aug-2018
Published online on: 10-09-2018

---

Plagiarism Check: Yes
Reviewed by: Dheeraj Nagore
Second Review by: Arun Singh
Final Approval by: Dr Ayush Dogra

---

How to Citeclose    |   Publication History close

 Views: (Visited 405 times, 1 visits today)    PDF Downloads: 523

Rafi Abdul-Majeed Al-Razzuqi¹, Ahmad Rahma Abu-Rageef², Wesal Sami Mehasin³ and Thulfaqar Rafi Abdul-Majeed Al-Razzuqi⁴

¹Department of Medical techniques, Al-Ma'moon University College, Baghdad, Iraq.

²Department of Pharmacology and Therapeutics, Al-Nahrain College of Medicine, Baghdad, Iraq.

³Department of Clinical Pharmacy, Al-Ma'moon University College, Baghdad, Iraq.

⁴Department of Medical techniques, Al-Yarmouk University College, Baghdad, Iraq.

Corresponding Author E-mail: rafialmajeed@yahoo.com

DOI : https://dx.doi.org/10.13005/bpj/1490

Abstract

Gastric ulcers result from an imbalance between endogenous defense mechanisms and certain aggressive agents. Many drugs were used to overcome this imbalance, but few literatures made on plants. Therefore, we try to evaluate the gastroprotective efficacy of two nutritional supplements (Vanadyl sulfate and Lycopene) in comparison to Lansoprazole.  Five groups of seven healthy albino male rats each were received an oral daily dose of above agents for ten days. Then 1.25 ml of 95% ethanol orally used to induce mucosal injury and animals were sacrificed 1 hour later. Glutathione and malondialdehyde were estimated. A significant elevation in glutathione level found in Vanadyl and Lycopene-received groups in comparison to lansoprazole-received group (717.13±19.47 μmol/gm wet tissue, 609.55±17.6 μmol/gm wet tissue and 512.07±25.32 μmol/gm wet tissue respectively), with a significant reduction in malondialdehyde level (10.63±0.92 nmol/gm wet tissue, 12.66±0.56 nmol/g wet tissue and 14.90±0.33 nmol/gm wet tissue respectively). This revealed gastro-protective effects of Vanadyl and Lycopene in ameliorating the oxidative cellular damage.

Keywords

Malondialdehyde; Gastro-protective; lansoprazole-Received

Introduction

Gastric mucosal injury generally results from an imbalance between the corrosive action of acid-pepsin and the mucosal integrity which is maintained by endogenous defense mechanisms such as glutathione (GSH), gastric mucus, bicarbonate secretion and prostaglandins.¹ This imbalance may also be caused by Helicobacter pylori,² starvation³ and ingestion of some ulcerogenic drugs like aspirin,⁴ indomethacin,⁵ and ethanol.⁶ These aggressive factors lead to (1) significant gastric mucosal damage (2) decreased tissue glutathione levels (the major cellular antioxidant that prevents cell from damage)⁷ (3) increased malondialdehyde levels (a product of the decomposition of polyunsaturated fatty acids during cellular oxidative stress)⁸  (4) increased myeloperoxidase enzyme (MPO) levels.⁹ According to traditional medicine, Ibn Senna; the well-known and greatest philosopher used some food to relieve stomach aches. Nowadays, the ingredients of this food are known as nutritional supplements such as vanadyl sulfate and lycopene. Therefore, it is of interest to evaluate the protective effects of these two nutritional supplements in comparison with lansoprazole in ethanol-induced gastric lesion model in albino rat. Vanadyl sulfate (the oxidative form of the trace element vanadium) is a popular muscle building product, produces a significant anabolic effect through promoting muscle uptake of glucose¹⁰ and is found in eggs, carrots, soybeans and oats. Lycopene is a bright red, fat-soluble carotenoid pigment and is found in tomatoes, watermelon, papaya, and red guava. It exerts an anti-mutagenic effect via reducing the oxidative damage to DNA in humans.¹¹

Methods

Materials

They were bought from Al-Madinah known drug bureau (lansoprazole was procured from Actavis /Barnstaple/UK, Vanadyl sulfate from Aldniuds Co./Germany, and lycopene from Liptis Pharmaceuticals /USA). The kits for biochemical estimations were procured from Fluka /Switzerland, Sigma /St.Louis / USA and from Biolabs SA/Maizy/France.

Acute Toxicity Studies

They were carried out according to (1) the LD₅₀ of 95% ethanol was 6 ml/kg p.o.¹² (2) the safe American recommended dose of oral vanadyl sulfate is 5 mg/kg/day ¹³ and (3) lycopene has been given in doses as high as 2000mg/kg/day without any adverse effects.¹⁴ However, we decided 56% ethanol as 8g/kg once daily¹⁵ and 1/10^th of upper doses of vanadyl sulfate and lycopene to be considered for the experiments.¹⁶

Experimental Design

The protocol of the experiment was approved by Al-Nahrain College of Medicine/ Animal Ethics Committee (Approval No. AEC/31/16/CMANU).

Animals

Thirty-five healthy male albino rats (200-220 grams) supplied by Al-Nahrain College of Medicine, were used after five days of standard housing conditions. The rats were distributed randomly into five groups (n₌7) receiving a daily oral dose for ten days of :  1.5milliliter of distilled water for Group I as a negative control, 1.5milliliter of distilled water for Group II, lansoprazole 30mg/ kilogram body weight for Group III as a positive control,  vanadyl sulfate 0.5mg/ kilogram body weight for Group IV and finally lycopene 200mg/kilogram body weight for Group V .

On 11^th day (at 3.30 p.m), the animals fasted for 18 hours. At 9.30 a.m of next day, a single dose of 56% ethanol as 1.25milliter p.o was given to all rats except Group I. The animals were sacrificed 1 hour later and their stomachs were separated. Then, by a longitudinal incision, gastric mucosa was bared and washed with normal saline to be prepared for examinations.

Histological Study

After 48 hours of submerging in formalin, the specimens were desiccated, cleared and fixed in paraffin. The cut sections were stained with H&E dye to be examined under a polarized microscope.

Ulcer Index

The gastric specimens were laid flat and the lesions (in the form of hemorrhage or linear breaks on the glandular portion of gastric mucosa) were measured using a dissecting microscope (Hamburg/Germany; 10xs) with a square grid.¹⁷ Then the ulcer index of each specimen was calculated.¹⁸

Biochemical Study

Assay of Mucosal Glutathione¹⁹

After rinsing with cold saline, the specimen was sunken in alkaline solution (pH 8) for 5 minutes to maintain mucosal integrity. Then Acivicin was added to preserve glutathione activity. After centrifuging the sample, the supernatant was safeguarded at 4°C for 30 minutes during which 0.5 milliliter of trichloroacetic acid was added. By using spectrophotometer, glutathione level in each supernatant was measured.

Assay of Malondialdehyde²⁰

After rinsing with cold saline, the specimen was put in10 milliliter of potassium chloride (KCl) solution for 45 minutes to have a homogeneous solution. (0.5 milliliter) of this homogenate was added to a mixture of (Sodium dodecyl sulfate [0.2 milliliter]+ acetic acid [1.5 milliliter] + 2-thiobarbituric acid [1.5 milliliter] + distilled water [0.3 milliliter]). two minutes later, the whole mixture was incubated at 98°C for 1 hour. Then mixture was cooled in an ice-containing baker for 10 minutes. At this time, 5 milliliters of n-butanol: pyridine (15:1) was added. After centrifuging for 10 minutes, malondialdehyde level in each supernatant was measured using High Performance Liquid Chromatography (HPLC) with fluorescent detection based on 2-thiobarbituric acid assay.

Statistical Analysis

Data were analyzed using SPSS 13 software (IBM Corp., Armonk, N.Y., USA) as one-way ANOVA followed by student’s t-test. The results were reported as mean ± SEM. And P<0.05 was considered statistically significant.

Results

Induction of gastric lesions by ethanol 95% when was given orally in rats, was found to be approximately of 100% (group II). The obtained results from group IV (Vanadyl sulfate-received) and group V (lycopene-received) revealed significant elevation in GSH level (prevention index) as compared with group III (lansoprazole-received) and this equal to 75.38 ±2.66, 50.31± 4.78 and 91.06 ±0.25 respectively ( Table I). All tested and control drugs showed a high significant reduction in free radicals of gastric tissue extract through increasing GSH and decreasing MDA levels (Table II).

Table 1: Effect of lansoprazole, vanadyl sulfate and lycopene on ethanol -induced gastric ulcer parameters in rats.

Data are expressed as: mean ± Standard error of the mean, group no. =7. *P<0.05, **P<0.01. mm² ; millimeter square

Table 2: Effect of lansoprazole, vanadyl sulfate and lycopene on gastric lesion healing parameters in rats.

Data are expressed as: mean ± Standard error of the mean, group no. =7. *P<0.05, **P<0.01. GSH ; glutathione. MDA ; malondialdehyde. μmol/g; micromole per gram  nmol/g ; nanomole per gram.

Discussion

Gastric mucosal lesion is formed when aggressive factors go beyond the cellular self-defense mechanisms. This leads to an excessive generation of reactive oxygen species (ROS) that promote degradation of the epithelial membrane components causing mucosal damage to the acinar part of the stomach²¹ while the non-acinar portion remained relatively intact.²² Some natural substances for example Emblica Officinalis have been known to strengthen gastric defense in healing induced gastric ulcers, enhancing cellular detoxification mechanisms and repairing the damaged non-proliferating cells.²³ Therefore, our study evaluated two dietary supplements (vanadyl sulfate and lycopene) compared to Lansoprazole (a proton pump inhibitor) that has a significant gastric mucosa protecting effect. The results showed that vanadium sulphate and lycopene exhibit a gastro-protective effect on ethanol-induced ulcers by increasing the Prostaglandins E2 production which in turn increasing the reduced GSH levels that responsible with sticking to ROS for wash out.²²

Conclusion

Vanadium sulphate and lycopene, through their property of ROS suppression, appear to improve the destructive effects of ethanol on the gastric mucosa.

Acknoledgements

We like to express our appreciation to Professor Dr. Farooq Hassan Al-Jawad (Department of Pharmacology and therapeutics, Al-Yarmouk University College) for his valuable notes that enrich the study. Besides, Professor Dr. Abdul-Karim Al-Kufi (Department of Pharmacology and therapeutics, Al-Nahrain College of Medicine) for his help in funding agreement.

References

1. Laine L., Takeuchi K., Tarnawski A. Gastric mucosal defense and cytoprotection: bench to bedside. Gastroenterology. 2008;135:41–60.
   CrossRef
2. Sonnenberg A. Review article: historic changes of Helicobacter pylori-associated ulcers. Aliment Pharmacol Ther. 2013;38:329–42.
   CrossRef
3. Yamamoto T., Isono A., Mishina Y., et al. Gastroduodenal mucosal injury in patients taking low-dose aspirin and the role of gastric mucoprotective drugs: possible effect of rebamipide. J Clin Biochem Nutr. 2010;47:27–31.
   Crossref
4. Miura T., Muraoka S., Fujimoto Y. Lipid peroxidation induced by indomethacin with horseradish peroxidase and hydrogen peroxide: involvement of indomethacin radicals. Biochem. Pharmacol. 2002;63: 2069-74.
   CrossRef
5. Bujanda L. The effects of alcohol consumption upon the gastrointestinal tract. Am J Gastroenterol. 2000;95(12):3374-82.
   CrossRef
6. Chen-Road., HungandSu-Lin N. Acid-Induced Gastric Damage in Rats Is Aggravated by Starvation and Prevented by Several Nutrients. Biochemical Pharmacology. 2003;66(8):1499–503.
7. Halliwell B. Antioxidant characterization: methodology and mechanism. Biochemical Pharmacology. 1995;49:1341-8.
   CrossRef
8. Sener G.,Kapucu C.,Cetinel S., Cikler E., Ayanoğlu-Dülger G. Gastroprotective effect of leukotriene receptor blocker montelukast in alendronate-induced lesions of the rat gastric mucosa. Prostaglandins Leukot Essent Fatty Acids. 2005;72(1):1-11.
   CrossRef
9. Boden G., Chen X., Ruiz J., Van Rossum G. D.  Effects of vanadyl sulfate on carbohydrate and lipid metabolism in patients with non-insulin dependent diabetes mellitus. Metabolism: Clinic and Exper 1996;45(9):1130-5.
   CrossRef
10. Porrini1 M & Riso P. Lymphocyte Lycopene Concentration and DNA Protection from Oxidative Damage Is Increased in Women after a Short Period of Tomato Consumption. J. Nutr. 2000;130:189-92.
    CrossRef
11. Kulkarni S. K.  Experiments on intact preparations (in vitro and in vivo studies). Handbook of Experimental Pharmacology.3^rd edit. Delhi: Vallabh Prakashan. 1999;148-67.
12. Fawcett J. P., Villani P., Brichard S. M. Evaluation of Genotoxicity of Oral Vanadyl Sulfate in Vivo. Toxicology Letters. 2007;13:47-53
13. Jian W. Q., Liu Y. Z., Fukuda T. Twenty-eight-day oral toxicity study of lycopene from recombinant Escherichia coli in rats. Regul. Toxicol. Pharmacol. 2008;52:163-8.
    CrossRef
14. Park J. H., Jang K. J., Kim C. H., Kim J. H., et al. Ganoderma Lucidum Pharmacopuncture for Treating Ethanol-induced Chronic Gastric Ulcers in Rats. See comment in PubMed Commons below. J Pharma copuncture. 2015;18:72-8.
    CrossRef
15. Ahmad A., Gupta G., Afzal M., Kazmi I., Anwar F.  Antiulcer and antioxidant activities of a new extract from Morus alba. Life Sci 2015;92(3):202–10.
    CrossRef
16. Lee T. H., Lin C. C., Chung C. S., Lin C. K., Liang C. C., Tsai K. C. Increasing biopsy number and sampling from gastric body improve the sensitivity of rapid urease test in patients with peptic ulcer bleeding. Dig Dis Sci. 2015;60:454–457.
    CrossRef
17. Halland M., Young M., Fitzgerald M. N., Inder K., Duggan J. M., Duggan A.  Bleeding peptic ulcer: characteristics and outcomes in Newcastle, NSW. Intern Med J. 2011;41(8):605-9.
    CrossRef
18. Beutler E., Duron O and Kelly B. Reduced glutathione estimation. J Lb Clinical Med. 1996;61:82–6.
19. Ohkawa H.,Ohishi N., Yagi K.  Assay for lipid peroxides in animal tissues by thiobarbi-turic acid reaction. Anal. Biochem. 1979;95:351-8.
    CrossRef
20. Hidekazu S., Toshihiro N., Hitoshi T., Sachiko M., Toshifumi H. Roles of oxidative stress in stomach disorders. J Clin Biochem Nutr. 2012;50(1): 35–9.
21. Kenneth R. M.  Drugs Used in the Treatment of Gastrointestinal Diseases. Katzung’s Basic and Clinical Pharmacology 12^th ed. McGraw-Hill Companies. New York. 2012:1081-1114.
22. Vasanthkumar M., Parameswari R. P.,Kumar V. V.,Sangeetha M. K., Raghavendran H.B., Gayathri V., et al. Anti-ulcer role of herbo-mineral Siddha drug on experimentally induced gastric mucosal damage in rats. Hum Exp Toxicol. 2010;29(3):161-73.
    CrossRef
23. Okazaki M., Shimizu I., Ishikawa M., Fujiwara S., Yamamoto H., Shiraishi T., et al. Gastric mucosal levels of prostaglandins in patients with the gastric ulcer after treatment with lansoprazole in comparison to treatment with ranitidine. J Med Invest. 2007;54(12):83-90.
    CrossRef