|Year : 2021 | Volume
| Issue : 1 | Page : 6-10
Effect of renin-angiotensin-aldosterone system blockade on intestinal injury induced by indomethacin in rats
Asmaa Abdulaziz A. Rabee1, Yahya Saber E. Mansour2, Huda A Mariz3, Salwa Muftah Eljamay4
1 Department of Pharmacology, Faculty of Medicine, Omar Elmokhtar University, Derna, Libya
2 Department of Pharmacy and Toxicology, Faculty of Pharmacy, Omar Elmokhtar University, Elbieda, Libya
3 Department of Pharmaceutical Technology, College of Medical Technology, Derna, Libya
4 Department of Public Health Technology, College of Medical Technology, Derna, Libya
|Date of Submission||18-Nov-2020|
|Date of Acceptance||14-Mar-2021|
|Date of Web Publication||10-Apr-2021|
Dr. Salwa Muftah Eljamay
Department of Public Health Technology, College of Medical Technology, Derna
Source of Support: None, Conflict of Interest: None
Aim: The aim of this experimental study was to evaluate the possible cytoprotective effects of renin-angiotensin-aldosterone system (RAAS) blockade, captopril (10 mg/kg), telmisartan (10 mg/kg), and a proton-pump inhibitor (lansoprazole) on intestinal injury induced by indomethacin in rats. Materials and Methods: This effect was evaluated through the assessment of intestinal inflammatory biomarkers and oxidative stress parameters. Forty adult male albino rats weighing 170–200 g were used and divided equally into five groups. After the treatment, the following were assessed: Myeloperoxidase (MPO) enzyme activity, pro-inflammatory (interleukin-1beta [IL-1 β]) content and anti-inflammatory (IL-10) content, malondialdehyde (MDA) content, reduced glutathione (GSH) content, and superoxide dismutase (SOD) enzyme activity, and histopathological examination of the intestinal tissues was conducted. Results: Indomethacin group induced a significant increase in MDA content, also in inflammatory biomarker (MPO enzyme activity, IL-1 β content, and IL-10 content) compared to normal control, while indomethacin group induced a significant decrease in GSH content, SOD enzyme activity compared to normal control group. Captopril, telmisartan, lansoprazole administration before indomethacin-induced significant increase in GSH content, SOD enzyme activity, while induced significant decrease in MDA content as well as in inflammatory biomarker compared to indomethacin group, which indicate the antioxidant, anti-inflammatory effects, and the gastroprotective properties of RAAS blockers, which also shown in the histopathological examination of intestinal tissues. Conclusion: This study showed the possible antioxidant and anti-inflammatory effects as well as the gastroprotective properties of RAAS blockers. Telmisartan has more antioxidant and anti-inflammatory effect as well as cytoprotective action than captopril.
Keywords: Captopril, indomethacin, lansoprazole, renin-angiotensin-aldosterone system blockade, telmisartan
|How to cite this article:|
A. Rabee AA, E. Mansour YS, Mariz HA, Eljamay SM. Effect of renin-angiotensin-aldosterone system blockade on intestinal injury induced by indomethacin in rats. Libyan J Med Sci 2021;5:6-10
|How to cite this URL:|
A. Rabee AA, E. Mansour YS, Mariz HA, Eljamay SM. Effect of renin-angiotensin-aldosterone system blockade on intestinal injury induced by indomethacin in rats. Libyan J Med Sci [serial online] 2021 [cited 2021 Jul 24];5:6-10. Available from: https://www.ljmsonline.com/text.asp?2021/5/1/6/313532
| Introduction|| |
Peptic ulcer disease is a complex and multi-causal disease that occurs when biological balance between defense and aggressive factors in the gastrointestinal tract is disturbed. Among aggressive factors are endogenous factors such as gastric acid and pepsin secretion, active free radicals and oxidants, leukotrienes, and endothelins as well as exogenous factors such as ethanol or nonsteroidal anti-inflammatory drugs (NSAIDs). Indomethacin induces its gastrointestinal toxicity by interfere with mucosal cell regeneration through inhibition of prostaglandin-E2 (PGE2) synthesis, production of free radicals, reduction of gastric nitric oxide level, and invasion of activated neutrophils as well as induction of gastric cells apoptosis. Angiotensin II, a key peptide hormone in the renin-angiotensin-aldosterone system (RAAS), induces oxidative stress and inflammation. Telmisartan that can concurrently block the angiotensin II T1-receptor and activate peroxisome proliferator-activated receptor gamma (PPAR-gamma). PPAR-gamma, a ligand-dependent nuclear receptor, has been reported to exert a variety of pleiotropic effects, including anti-oxidative and anti-inflammatory effects, also has the potential to be implicated in the protection against gastric ulcer. Proton-pump inhibitors, a strong anti-secretary agent, have anti-inflammatory action beyond acid suppression agents that act on gastric (H+/K+) ATPase of parietal cells, have been extensively used for therapeutic control of acid-related disorders, including gastroesophageal reflux disease and peptic-ulcer diseases caused by stress, NSAIDs, and Helicobacter pylori infection.
Angiotensin II is involved in the several key steps of the inflammatory cascade that ultimately provokes intestinal injury and ulceration including polymorphonuclear leukocyte infiltration, probably, through the upregulation of adhesion molecules. The aim of the present experimental study is to investigate and compare the reffect of pretreatment with captopril and telmisartan on indomethacin-induced small intestinal injuries in rats.
| Materials and Methods|| |
Animals and experimental design
Forty adult male albino rats weighing 170–200 g were used and divided equally into five groups; each contained eight rats, in clean capacious macrolane cages under standard laboratory conditions, as the following: Group I: Control group. Group II: indomethacin was administered in a dose of 10 mg/kg i. p. for 3 days. Group III: Captopril was administered in a dose of 10 mg/kg p. o. for 14 days and indomethacin was administered in a dose of 10 mg/kg i. p. in last 3 days. Group IV: temisartan was administered in a dose of 10 mg/kg p. o. for 14 days and indomethacin was administered in a dose of 10 mg/kg i. p. in last 3 days. Group V: Lansoprazole was administered (as a reference drug) in a dose of 50 mg/kg, p. o. for 14 days and indomethacin was administered in a dose of 10 mg/kg i. p. in last 3 days. Experiments were conducted to study the effect of the drugs under investigation on indomethacin-induced small intestinal injury through measuring the following parameters:
Oxidative stress parameters
Malondialdehyde (MDA) content, reduced glutathione (GSH) content, and superoxide dismutase (SOD) enzyme activity.
Myeloperoxidase (MPO) enzyme activity, pro-inflammatory interleukin-1beta (IL-1 β) content, and anti-inflammatory (IL-10) content, in addition to histopathological examinations.
Drugs and chemicals
- Indomethacin vials, 50 mg, EL Nile Co. for pharmaceutical and chemical industries (Egypt)
- Captopril tablet, 25 mg, Epico (Egypt)
- Telmisartan tablets, 80 mg (Hikma Pharmaceuticals USA Inc.)
- Lansoprazole capsules 30 mg, Sedico (Egypt)
- Tween 80 (Sigma-Aldrich): It was used for the emulsification of captopril, telmisartan, and lansoprazole. All other chemicals used were of good quality and analytical grade.
MPO activity, Pro-inflammatory IL-1 β content and anti-inflammatory (IL-10) content measured using a rat-specific enzyme-linked immunosorbent assay kits.
All animals were scarified by cervical dislocation and their small intestine were quickly excised and washed with saline and used for the preparation of tissue homogenate 20% (in 0.9% NaCl using homogenizer (X620 CAT, made in Germany) and stored at − 80°C until analysis. The small intestine of two animals from each group was kept in 10% formalin and used for histopathology examination.
The animals were handled according to the guidelines of local ethical committee which comply with the international laws for use and care of laboratory animals. Animal Ethics Committee Approval No. Ethics/research/OMU/2020-87.
Determination of oxidative stress parameters
The antioxidant parameters were determined in small intestine homogenate (20%) after centrifugation of tissues at 3.000 rpm for 10 min, and the supernatant was used.
Determination of malondialdehyde content
The colorimetric determination of Thiobarbituric acid reactive substances (TBARS) content is based on the reaction of one molecule of MDA with two molecules of thiobarbituric acid in acidic medium at the temperature of 95°C for 45 min. The resultant pink pigment product is extracted by n-butanol, and then its absorbance is determined at 535 nm and 520 nm. The difference in absorbance between the two determinations was used to calculate the TBARS value.
Determination of reduced glutathione content
Determination of nonprotein sulfhydryl compounds (indicative to GSH content) was done according to the method of Ellman. The method depends on the reduction of Ellman's reagent (5, 5'-dithio-bis (2-nitrobenzoic acid) by SH group in GSH to form an intense yellow product (5–thio-2 nitrobenzoic acid) which can be measured colorimetrically at 412 nm.
Determination of superoxide dismutase activity
SOD activity in the homogenate was determined by the difference between auto-oxidation of pyrogallol alone and in the presence of homogenate that contain SOD.
Determination of inflammatory biomarkers
- Determination of MPO enzyme activity (ELISA kit CAT. NO. SEA601Ra)
- Determination of IL-1 β content (ELISA kit CAT. NO. EL10028)
- Determination of IL-10 content (ELISA kit CAT. NO. ELR-IL10-001C).
Statistical analysis was done using the computer program (prism 5). The quantitative data were presented in the form of mean ± standard error of the mean. Statistical analysis of the difference between the groups was performed using the one–way analysis of variance followed by Tukey-Kramer test for multiple comparisons.
| Results|| |
Indomethacin group, induced a significant increase in MDA content compared to normal control group, while pretreated of captopril-induced significant decrease in MDA content compared to indomethacin group as well as with telmisartan, and with lansoprazole indomethacin group induced a significant decrease in GSH content, SOD enzyme activity compared to normal control group.
Captopril, telmisartane, lansoprazole administration before indomethacin induced significant increase in GSH content, SOD enzyme activity compared to indomethacin group. Indomethacin group induced a significant increase in inflammatory biomarker (MPO enzyme activity, IL-1 β content and IL-10 content compared to normal control while decreased with other pretreated groups [Table 1] and [Figure 1].
|Table 1: Results of biochemical tests among the four groups and compare with control group|
Click here to view
Histopathological examination of rat intestinal tissues in the four groups (i.e., indomethacin group [Figure 2], captopril and indomethacin group, valsartan and indomethacin group, and lansoprazol and indomethacin group), as compared with control group showed inflammatory cells infiltration in the lamina propria of the villi with diffuse goblet cells formation in the lining mucosal epithelium and severe congestion in the blood vessels in submucosa.
|Figure 2: Transverse section of rat small intestine in indomethacin group showing inflammatory cells infiltration in the lamina propria of the villi with diffuse goblet cells formation in the lining mucosal epithelium|
Click here to view
| Discussion|| |
NSAIDs such as indomethacin are extensively recommended in the management of pain, fever, and inflammation while its gastrointestinal tract adverse effects disturb its use. Indomethacin induces its gastrointestinal toxicity through several mechanisms such as an increase in gastric acid secretion; interfere with mucosal cell regeneration through inhibition of PGE2 synthesis, production of free radicals, reduction of gastric nitric oxide level, and invasion of activated neutrophils as well as induction of gastric cells apoptosis. Many previous studies have been done by damaging effect of indomethacin on gastrointestinal mucosa., The present study revealed that indomethacin-induced duodenal ulcer through increase gastric juice volume, free and total acidity as well as a decrease in gastric protective prostaglandin. Pretreatments with captopril or talmisartane have ability to decrease gastric acid secretion induced by indomethacin. Gastric secretion is a controlled by captopril or telmisartan is characterized by the property of stimulating PPARgamma. Telmisartan reduced the ulcer index in both ulcer models. These results are supported by the findings of Nakagiri et al.
Indomethacin group, induced a significant increase in MDA content compared to normal control group, which indicates that tissues exposed to oxidative stress include large amounts of toxic oxygen radicals, which induce lipid peroxidation leading to MDA formation., The lowest MDA values were detected in the groups treated by captopril, telmisartan, and lansoprazole compared with indomethacin group, there was an inhibition in SOD as well as GSH content in indomethacin treated rats compared to the corresponding control. The decline in SOD and GSH level in the intestinal mucosa of rats treated with indomethacin hence may render the mucosa more susceptible to injury by indomethacin. Thus, the inhibition of antioxidants leads to the accumulation of ROS., On the other hand, pretreated of rats with captopril, telmisartan, lansoprazole before indomethacin induced significant increase in GSH content, SOD enzyme activity compared to indomethacin group which indicates the cytoprotective activity of these drugs.
Indomethacin group induced a significant increase in inflammatory biomarker (MPO enzyme activity, IL-1 β content, and IL-10 content compared to normal control. Indomethacin prevents synthesis of PGE2, which is the main factor that involves in gastric mucosal defense mechanism through many of its actions. First, it regulates gastric blood flow to maintain the level of mucus and bicarbonate secretion as these two substances are secreted in flow-dependent fashion. Furthermore, endogenous PGE2 also controls the epithelial cell proliferation and restitution, mucosal immunocyte function, and basal acid secretion, also increased oxygen-free radicals increases the protease secretion, and capillary blood flow obstruction which will result in gastric injury., Besides interfering of PGE2 pathway, NSAIDs also raise the nitric oxide synthase activity which causes increase in pro-inflammatory cytokine and damage of intestinal mucosa while decreased with captopril, and telmisartan groups which indicate anti-inflammatory and cytoprotective effect of RAAS blockade. Angiotensin II, through the binding with angiotensin receptor, is also capable of promoting inflammation indirectly through the stimulation of several transcription factors, also Angiotensin II stimulates the production of proinflammatory cytokines and chemokines, such as IL-6, IL-8, which is an important mediator for the direct migration of monocytes.
The results of telmisartan is nearly to the result of standard gastroprotective (lansoprazole), which indicates that telmisartan has better antioxidant and cytoprotective properties than theirs of captopril. In histopathological examination, we found lymphoid hyperplasia in the follicles of the submucosal layer associated with inflammatory cells infiltration in the mucosa and diffuse goblet cells formation in the lining mucosal epithelium of the villi in indomethacin group, but in pretreating groups of telmisartan, captopril, and lansoprazole, we found diffuse goblet cells formation in the lining mucosal epithelium of the villi which indicate anti-inflammatory and cytoprotective action of RAAS blockade.
| Conclusion|| |
This study showed the possible antioxidant and anti-inflammatory effects as well as the gastroprotective properties of RAAS blockers. Telmisartan has more antioxidant and anti-inflammatory effect as well as cytoprotective action than captopril.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Sumbul S, Ahmad MA, Mohd A, Mohd A. Role of phenolic compounds in peptic ulcer: An overview. J Pharm Bioallied Sci 2011;3:361-7.
Borrelli F, Izzo AA. The plant kingdom as a source of anti-ulcer remedies. Phytother Res 2000;14:581-91.
Wallace JL, McKnight GW, Keenan CM, Byles NI, MacNaughton WK. Effects of leukotrienes on susceptibility of the rat stomach to damage and investigation of the mechanism of action. Gastroenterology 1990;98:1178-86.
Soll A, Graham D. Peptic ulcer disease. In: Yamada T, editor. Text Book of Gastroenterology. 5th
ed. USA: Blackwell Publication Ltd.; 2009. p. 936-41.
Takeuchi K. Pathogenesis of NSAID induced gastric damage: Importance of cyclooxygenase inhibition and gastric hypermotility. WJG 2012;18:2147-60.
Matsui H, Shimokawa O, Kaneko T, Nagano Y, Rai K, Hyodo I. The pathophysiology of non-steroidal antiinflammatory drug (NSAID)-Induced mucosal injuries in stomach and small intestine. J Clin Biochem Nutr 2011;48:107-11.
Welch WJ. Angiotensin II-dependent superoxide: Effects on hypertension and vascular dysfunction. Hypertension 2008;52:51-6.
Benson SC, Pershadsingh HA, Ho CI, Chittiboyina A, Desai P, Pravenec M, et al
. Identification of telmisartan as a unique angiotensin II receptor antagonist with selective PPARgamma-modulating activity. Hypertension 2004;43:993-1002.
Takano H, Komuro I. Peroxisome proliferator-activated receptor gamma and cardiovascular diseases. Circ J 2009;73:214-20.
Fellenius E, Berglindh T, Sachs G, Olbe L, Elander B, Sjöstrand SE, et al
. Substituted benzimidazoles inhibit gastric acid secretion by blocking (H++K+) ATPase. Nature 1981;290:159-61.
Langtry HD, Wilde MI. Omeprazole. A review of its use in Helicobacter pylori
infection, gastro-oesophageal reflux disease and peptic ulcers induced by nonsteroidal anti-inflammatory drugs. Drugs 1998;56:447-86.
Yamamoto A, Itoh T, Nasu R, Nishida R. Sodium alginate ameliorates indomethacin-induced gastrointestinal mucosal injury via inhibiting translocation in rats. World J Gastroenterol 2014;20:2641-52.
Sulpizio AC, Pullen MA, Edwards RM, Brooks DP. The effect of acute angiotensin-converting enzyme and neutral endopeptidase 24.11 inhibition on plasma extravasation in the rat. J Pharmacol Exp Ther 2004;309:1141-7.
Morsy M, Ashour O, Amin E, Rofaeil R. Gastroprotective effects of telmisartan on experimentally-induced gastric ulcers in rats. Pharmazie 2009;64:590-94.
Patil AS, Singh AD, Mahajan UB, Patil CR, Ojha S, Goyal SN. Protective effect of omeprazole and lansoprazole on β-receptor stimulated myocardial infarction in Wistar rats. Mol Cell Biochem 2019;456:105-13.
Mihara M, Uchiyama M. Determination of malonaldehyde precursor in tissues by thiobarbituric acid test. Anal Biochem 1978;86:271-8.
Ellman GL. Tissue sulphydryl groups. Arch Biochem Biophys 1959;82:70-7.
Marklund SL. Pyrogallol autooxidation. In: Handbook of Methods for Oxygen Radical Research. Boca Raton, Florida: C.R.C Press; 1985. p. 243-7.
Abdel-Raheem IT. Gastroprotective effect of rutin against indomethacin-induced ulcers in rats. Basic Clin Pharmacol Toxicol 2010;107:742-50.
Cheng YT, Lu CC, Yen GC. Phytochemicals enhance antioxidant enzyme expression to protect against NSAIDinduced oxidative damage of the gastrointestinal mucosa. Mol Nutr Food Res 2017;61:1-9.
Ibrahim FM, Attia HN, Aly Maklad YA, Ahmed KA, Ramadan MF. Biochemical characterization, anti-inflammatory properties and ulcerogenic traits of some cold-pressed oils in experimental animals. Pharm Biol 2017;55:740-8.
Nakagiri A, Sunamoto M, Murakami M. Angiotensin AT1 receptor blockers suppress ischemia/reperfusion-induced gastric injury in rats. Inflammopharmacology 2007;15:171-4.
Talas DU, Nayci A, Polat G, Atis S, Comelekoglu U, Bagdatoglu OT, et al
. The effects of dexamethasone on lipid peroxidation and nitric oxide levels on the healing of tracheal anastomoses: An experimental study in rats. Pharmacol Res 2002;46:265-71.
Bilici M, Ozturk C, Dursun H, Albayrak F, Saglam MB, Uyanik A, et al
. Protective effect of mirtazapine on indomethacin-induced ulcer in rats and its relationship with oxidant and antioxidant parameters. Dig Dis Sci 2009;54:1868-75.
Othman AI, El-Missiry MA, Amer MA. The protective action of melatonin on indomethacin-induced gastric and testicular oxidative stress in rats. Redox Rep 2001;6:1-5.
Ito M, Suzuki Y, Ishihara M, Suzuki Y. Anti-ulcer effects of antioxidants: Effect of probucol. Eur J Pharmacol 1998;354:189-96.
Hawkey CJ. Nonsteroidal anti-inflammatory drug gastropathy. Gastroenterology 2000;119:521-35.
Wallace JL. Nonsteroidal anti-inflammatory drugs and gastroenteropathy: The second hundred years. Gastroenterology 1997;112:1000-16.
Appleyard CB, McCafferty DM, Tigley AW, Swain MG, Wallace JL. Tumor necrosis factor mediation of NSAID-induced gastric damage: Role of leukocyte adherence. Am J Physiol 1996;270:G42-8.
Rainsford KD. Microvascular injury during gastric mucosal damage by anti-inflammatory drugs in pigs and rats. Agents Actions 1983;13:457-60.
Funakoshi Y, Ichiki T, Shimokawa H, Egashira K, Takeda K, Kaibuchi K, et al
. Rho-kinase mediates angiotensin II-induced monocyte chemoattractant protein-1 expression in rat vascular smooth muscle cells. Hypertension 2001;38:100-4.
[Figure 1], [Figure 2]