Possible protective role of verapamil on ischemia/reperfusion induced changes in the jejunal mucosa of adult male albino rat: Histological and biochemical study
DOI:
https://doi.org/10.56511/JIPBS.2018.v5.i01.pp92-102Abstract
Background: The small intestine is extremely sensitive to ischemia-reperfusion (I/R) injury and a range of microcirculatory disturbances which contribute to tissue damage. Previous studies have shown that verapamil plays an important physiological role in the microvascular environment. This study aimed to evaluate the protective effects of verapamil in ischemia/reperfusion (I/R) – induced mucosal injury in the jejunum. Fifty adult albino rats were divided into five groups: group I (Control untreated); group II (Sham-operated); group III (sham-operated +verapamil); group IV (Ischemia/Reperfusion) and Group V (I/R + verapamil). At the end of the reperfusion period the jejunum was extracted and prepared for histological and immunohistochemical examination. Distribution of myeloperoxidase (MPO) stained cells and the levels of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH,) were also determined in all dissected tissues. Morphometric study was also done and statistical results were analyzed. Intestinal IR caused severe intestinal mucosa injury especialy epithelial cell damage. Group (I/R) revealed villus structural alterations, cellular infiltration and hemorrhage. MPO and goblet cells showed a significant increase in the number with elevation of oxidative stress marker. Co-administration of verapamil ameliorated these histological changes and significantly decreased mucosal damages compared to group I/R. Conclusion: Verapamil can protect against intestinal I/R probably through inhibition of oxidative stress and neutrophil infiltration. So it can be used safely in mesenteric occlusive diseases, since it induces improvement of circulation and relief of concomitant structural changes.
Keywords:
Intestine, Verapamil, I/R, oxidative stressDownloads
References
Buraimoh AA: Effect of aluminium chloride exposure on the histology of small intestine of wistar rats. Int j of Appl. Sci. and Tech. 2012; 2(10):123-132
Clark JA and Coopersmith CM: Intestinal crosstalk: a new paradigm for understanding the gut as the ‘motor’ of critical illness . Shock 2007; 28:384-393.
Sun Y, Xu Y, and Wang GN: Pterostilbene prevents intestinal ischemia reperfusion injury in Wistar rats via modulation of antioxidant defense and inflammation. Trop. J Pharm. Res. 2015; 14(8): 1383-1391.
Pierro A and Eaton S: Intestinal ischemia reperfusion injury and multisystem organ failure. Semin. Pediatr. Surg. 2004;13:11 – 17.IR---
Cerqueira NF, Hussni CA and Yoshida WB: Pathophysiology of mesenteric ischemia/reperfusion: A review. Acta Cir. Bras. 2005; 20:336 – 343.
Koike K, Moore FA, Moore EE, Read RA, Carl VS and Banerjee A: Gut ischemia mediates lung injury by a xanthine oxidase dependent neutrophil mechanism. J Surg. Res. 1993; 54(5): 469-473.
Shirasugi N, Wakabayashi G, Shimazu M, et al.: Up-regulation of oxygen-derived free radical by interleukin-1 in hepatic ischemia/reperfusion injury. Transplantation 1997; 64(10): 1398-1403.
Sagiroglu T, Sezer A and Altaner S: The Effects of Sirolimus on Target Organs During Mesenteric Ischemia and Reperfusion Damage in an Experimental Rat Model. Current Therapeutic Research 2011; 72(2):7993.
Granger DN, Hollwarth ME and Park DA: Ischemia reperfusion injury: role of oxygen-derived free radicals. Acta Physiol Scand 1986; 548:47 – 56.
Sukhotnik I, Brod V and Lurie M et al.: The effect of 100% oxygen on intestinal preservation and recovery following ischemia-reperfusion injury in rats. Crit Care Med. 2009; 37:1054 – 1061.
Carden DL and Granger DN: Pathophysiology of ischaemia-reperfusion injury. J Pathol. 2000; 190:255 – 266.
Jun P, Ko N and English J et al.: Endovascular treatment of medically refractory cerebral vasospasm following aneurysmal subarachnoid hemorrhage. Am. J. Neuroradiol 2010; 31: 1911 – 6.
Bell TJ, et al.: Cell-specific alternative splicing increases calcium channel current density in the pain pathway. Neuron 2004; 41(1):127 – 138.
Qian AH, et al.: Voltage-gated potassium channels in IB4-positive colonic sensory neurons mediate visceral hypersensitivity in the rat. Am J Gastroenterol. 2009; 104(8):2014 – 2027.
Tucci S, Borelli-Bovo T, Cologna A, Tiraboschi B, Martins A and Roselino J: Calcium channel blocker and renal mitochondrial function in warm renal ischemia. Int. Braz. J Urol. 2005; 31: 1677 – 6119.
Dosluoglu HH, Aktan AO, Yegen C, Okboy N, Yalcm AS and Yahn R. et al.: The cytoprotective effects of verapamil and iloprost (ZK 36374) on ischemia/ reperfusion injury of kidneys. Transpl. Int. 1993; 6: 13842.
Qian A, SongD, Li Y, Liu X, Tang D, Yao W and Yuan Y: Role of voltage gated Ca2+ channels in rat visceral hypersensitivity change induced by 2,4,6-trinitrobenzene sulfonic acid Molecular pain 2013; 9(15):1-10.
Osman AS, Osman AH and Kamel MM: Study of the protective effect of ischemic and pharmacological preconditioning on hepatic ischemic reperfusion injury induced in rats. Journal of gastroenterology and hepatology 2017; 1:105-111.
Teke Z, Kabay B, Ozden A, Yenisey C, Bir F, Demirkan NC, Bicakci T and Erdem E: Effects of tempol, a membrane permeable radical scavenger, on local and remote organ injuries caused by intestinal ischemia/reperfusion in rats. J. Surg. Res. 2008; 149(2):259-271.
Messiha B and Abo-Youssef A: Protective effects of fish oil, allopurinol, and verapamil on hepatic ischemia-reperfusion injury in rats. J. Nat. Sci. Biol. Med. 2015; 6: 351 – 5.
Fujino Y, Suzuki Y, Kakinoki K, Tanioka Y, Ku Y and Kuroda Y: Protection against experimental small intestinal ischaemia – reperfusion injury with oxygenated perfluorochemical. British Journal of Surgery 2003; 90: 1015 – 1020.
Bancroft J and Layton C: Hematoxylin and eosin. In: Suvarna SK, Layton C and Bancroft JD (ed). Theory and Practice of histological techniques. 7th edition. Churchill Livingstone of El Sevier, Philadelphia 2013; 172214.
Aban, N., Cinel, L., Tamer, L., Aktas, A. and Aban, M: Ischemic preconditioning reduces caspase-related intestinal apoptosis. Surg. Today 2005; 35:228-34.
Onal O, Yetisir F, Sarer ES, Zeybek ND, Onal CO, Yurekli B, Celik HT, Sirma A, and Mehmet K: Prophylactic Ozone Administration Reduces Intestinal Mucosa Injury Induced by Intestinal Ischemia-Reperfusion in the Rat. Mediators of inflammation 2015; 2015:1-8.
Kurokawa T, Itagaki S, Yamaji T, Nakata C, Noda T, Hirano T and Iseki K: Antioxidant activity of a novel extract from bamboo grass (AHSS) against ischemia-reperfusion injury in rat small intestine. Biol. Pharm. Bull. 2006; 29(11):2301-2303.
Sun Y, Oberley LW and Li Y: A simple method for clinical assay of superoxide dismutase. Clin Chem.1988; 34: 497-500.
Nishikimi, M.; Appaji, N. and Yagi, K: The occurrence of superoxide anion in the reaction of reduced phenazine methosulfate and molecular oxygen'. Biochem. Biophys. Res. Commun. 1972; 46:849-54.
Rotruck JT, Pope AL, Ganther HE, Swanson AB, Hafeman DG and Hoekstra WG: Selenium: biochemicalrole as a component of glutathione peroxidase. Science 1973; 79:588-590.
Jefferies H, Coster J, Khalil A, Bot J, McCauley RD and Hall JC: Glutathione. ANZ J. Surg. Jul. 2003; 73(7):517-522.
Kumral A, Gonenc S, Acikgoz O, Sonmez A, Genc K, Yilmaz O, Gokmen N, Duman N and Ozkan H: Erythropoietin increases glutathione peroxidase enzyme activity and decreases lipid peroxidation levels in hypoxic-ischemic brain injury in neonatal rats. Biol. Neonate 2005; 87(1):15-18.
Frank T, Grulke S, Deby-Dupont G, Deby C, Duvivier H, Peters F and Serteyn D: Development of an enzyme linked immunosorbent assay for specific equine neutrophil myeloperoxidase measurement in blood. J Vet. Diagn. Invest. 2005; 17:412 – 419.
Grisham M, Benoit J and Granger D: Assessment of leukocyte involvement during ischemia and re-perfusion of intestine. Meth. Enzymol.1990; 186:729 – 742.
Junqueira LC and Carneiro J: Basic histology. 11th Ed. USA, McGrawHill. 2005. pp.298311.
Wilson C, Gupta R, Gilmour, DG and Imrie, CW: Acute superior mesenteric ischaemia. Br. J Surg. 1987;74: 279 – 81.
Oldenburg WA, Lau LL, Rodenberg TJ, Edmonds HJ and Burger, CD: Acute mesenteric ischemia: A clinical review. Arch Int. Med. 2004; 164: 1054 – 62.
Levy PJ, Krausz MM and Manny J: Acute mesenteric ischemia: improved results – a retrospective analysis of ninety-two patients. Surgery 1990; 107:372-380.
Benjamin E, Oropello JM and Iberti TJ: Acute mesenteric ischemia: pathophysiology, diagnosis, and treatment. Dis. Mon.1993; 39: 131 – 210.
Berland T. and Oldenburg WA: Acute mesenteric ischemia. Curr. Treat Options Gastroenterol. 2008; 11:3-10.
Rijke RP, Van Der Meer-Fieggen W and Galjaard H: Effect of villus length on cell proliferation and migration in small intestinal epithelium. Cell Tissue Kinet.1974; 7: 577-86.
Rocha BC, Mendes RR, Lima GV, Albuquerque GS, Araújo LL, Jesus MN, Santos WL and Carreiro MC: Experimental model of mesenteric ischemia/reperfusion by abdominal aorta clamping in Wistar rats. Rev. Col. Bras. Cir. 2012; 39(3): 207-210.
Toth S, Jonecova Z, Curgali K, Maretta M, Soltes J, Savana M, Kalpadikis T, Caprenda M, Adamek M, Rodrigo L and Kruzliak P: Quercetini attenuates the ischemia reperfusion induced COX2 and MPO expression in the small intestine mucosa. Biomedicine and Pharmacotherapy 2017; 95:346-354.
Grootjans J, Lenaerts K, Derikx JP, Matthijsen RA, de Bruine AP and van Bijnen AA. et al.: Human intestinal ischemia-reperfusion-induced inflammation characterized: experiences from a new translational model. Am. J Pathol. 2010; 176(5):2283-91.
Yamamoto S, Tanabe M, Wakabayashi G, Shimazu M, Matsumoto K and Kitajima M: The role of tumor necrosis factor-alpha and interleukin-1beta in ischemia reperfusion injury of the rat small intestine. J. Surg. Res. 2001; 99:134-41.
Vincenti M, Behrends M, Dang K, Park YH, Hirose R and Blasi-Ibanez A. et al.: Induction of intestinal ischemia reperfusion injury by portal vein outflow occlusion in rats'. J. Gastroenterol. 2010; 45:1103-10.
Chiu C J, McArdle A H, Brown R, Scott H J and Gurd F N: Intestinal mucosal lesion in low-flow states. I. A morphological, hemodynamic, and metabolic reappraisal. Arch. Surg.1970; 101:478-83.
Uygun M, Yilmaz S, Pekdemir M, Duman C and Gürbüz YS: The diagnostic value of ischemia-modified albumin in a rat model of acute mesenteric ischemia. Acad. Emerg. Med. 2011; 18(4):355-9.
Silva FN: Normothermic hepatic ischemia in rats: study of cellular injury by the use of continuous and intermittent pedicular clamping. Rev Col Bras Cir. 2002; 29(6):342-8.
Tang ZH, Qiang JW, Feng XY, Li RK, Sun RX and Ye XG: Acute mesenteric ischemia induced by ligation of porcine superior mesenteric vein: multidetector CT evaluations. Acad Radiol. 2010; 17(9):1146-52.
Juel IS, Solligård E, Lyng O, Strømholm T, Tvedt KE and Johnsen H. et al.: Intestinal injury after thoracic aortic cross-clamping in the pig. J Surg Res. 2004; 117(2):283-95.
Brasileiro JL, Inoye CM, Aydos RD, Silva IS, Falcao GR, Marks G and Pereira DM: Ischemia and reperfusion of rat small intestine using pentoxyfilline and prostaglandin E1, Acta Cirurgica Brasileira 2013; 28(11):767-773.
Jia Z, Lian W, Shi H, Cao C, Han S, Wang K, Li M and Zhang X: Ischemic precondioning protects against intestinal ischemia/reperfusion injury via the HIF-1 α / miR-21 axis, Scientific reports 2017; 7:1-11
Isik A, Peker K, Gursul C, Sayar I, Firat D, Yilmaz I and Demiryilmaz I: The effect of ozone and naringin on intestinal ischemia/reperfusion injury in an experimental model. International journal of surgery 2015; 21:3844.
Sola A, Hotter G, Prats N, Xaus C, Gelpi E and Roselló-Catafau J: Modification of oxidative stress in response to intestinal preconditioning. Transplantation 2000; 69(5):767-72.
Santos CH, Gomes OM, Pontes JV, Miiji LN and Bispo MA: The ischemic preconditioning and postconditioning effect on the intestinal mucosa of rats undergoing mesenteric ischemia/reperfusion procedure. Acta cir bras. 2008; 23(1):22-8.
Dupre LD, Vallee M, Bossard C, Laboisse CL and Jarry A: A multiparametric approach to monitor the effect of Y-secretase inhibition along the whole intestinal tract. Disease Models & Mechanisms 2012; 5:107-114.
Menke V, van Es JH, de Lau W, van den Born M, Kuipers EJ, Siersema PD, de Bruin RW, Kusters JG and Clevers H: Conversion of metaplastic Barrett’s epithelium into post -mitotic goblet cells by gamma-secretase inhibition.Dis. Model. Mech. 2010; 3: 104-110.
Sekine A, Akiyama Y, Yanagihara K and Yuasa Y. Hath1 up-regulates gastric mucin gene expression in gastric cells. Biochem. Biophys. Res. Commun.2006; 344: 1166-1171.
Matovelo JA, Sund RB and Landsverk T: Morphological and functional recovery following exposure to deoxycholic acid. APMIS 1989; 97:798 – 810.
Ikeda H, Yang C, Tong J, Nishimaki H, Masuda K, Takeo T, Kenji K and Itoh G: Rat small intestine goblet cell kinetics in the process of restitution of surface epithelium subjected to ischemia reperfusion injury. Digestive diseases and science 2002; 47(3):590-600.
Luo C, Laine VJ, Ylinen L, Teros T, Mäkinen M, Ristimäki A and Simell O: Expression of cyclooxygenase-2 in intestinal goblet cells of prediabetic NOD mice. Acta Physiol. Scand. 2002; 174: 265 – 267.
Gomes OM, Brasileiro Filho G, Salles RLA, Linhares RE, Nazar MC, Porto LAB, Paixão RM and Miranda MH: Ischemia-reperfusion histopathology alterations of the rabbit intestinal wall with and without ischemic preconditioning. Acta Cir Bras. 2011; 26(4):285-8.
Bertoletto PR, Chaves JC, Fagundes ATN, Simões RS, Oshima CTF, Simões MJ and Fagundes DJ: Effect of different periods of hyperbaric oxygen on ischemia-reperfusion injury of rat small bowel. Acta Cir Bras. 2008; 23(1):23-6.
Francisco Neto A, Silva JCCB, Fagundes DJ, Percário S, Novos F, Juliano Y and Moreira Neto AA: Study of oxidative changes of total antioxidant capacity of nitric oxide in rats submitted to ischemia and reperfusion of hind limbs. Acta Cir Bras. 2005; 20(2):134-9.
Topcua I, Vatanseverb S, Varc A, Cavusb Z, Cilakera S and Sakarya M; The effect of Misoprostol, a prostaglandin E1 analog, on apoptosis in ischemia – reperfusion-induced intestinal injury. Acta Histochem. 2007;109:322-9.
Guven A, Tunc T, Topal T, Kul M, and Korkmaz A.et al.: α -lipoic acid and ebselen prevent ischemia/reperfusion injury in the rat intestine. Surgery Today 2008; 38:1029-1035.
Cerqueira NF, Hussni CA and Yoshida WB: Pathophysiology of mesenteric ischemia/reperfusion: A review. Acta Cir. Bras. 2005; 20:336-
Powers KA, Zurawska J, Szaszi K., Khadaroo RG, Kapus A, and Rotstein OD: Hypertonic resuscitation of hemorrhagic shock prevents alveolar macrophage activation by preventing systemic oxidative stress due to gut ischemia/reperfusion. Surgery 2005; 137(1): 66 – 74,
Lu YQ, Cai XJ, Gu LH, Wang Q, Huang WD, and Bao DG: Early difference in apoptosis of intestinal mucosa of rats with severe uncontrolled hemorrhagic shock after three fluid resuscitation methods. Chinese Medical J. 2006; 119(10): 858 – 863.
Grotz MR, Deitch EA, Ding J, Xu D, Huang Q, and Regel G : Intestinal cytokine response after gut ischemia: role of gut barrier failure. Annals of Surgery 1999; 229 (4):478 – 486.
Liang JC, Chen HR, Chiu CC, Liou SF, Chen IJ, and Yeh JL: Protective effect of labedipinedilol-A, a novel dihydropyridine-type calcium channel blocker, on myocardial apoptosis in ischemia-reperfusion injury. Life Sciences 2006; 79(13):1248 – 1256.
Chiu WT, Wang YH, Tang MJ, and Shen MR: Soft substrate induces apoptosis by the disturbance of Ca2+ homeostasis in renal epithelial LLCPK1 cells. J Cellul. Physiol. 2007; 212(20): 401 – 410.
Amara WM, El-SiSi AE, AbuRisha SE, ElDesouky KI and Mostafa SA: Rosiglitaone protection against experimentally induced intestinal ischemia/reperfusion. Res. Pharmaceutical Biotechnol. 2012; 4(1):6-17.
Song WB, Wang YY, Meng FS, Zhang QH, Zeng JY, Xiao LP, Yu XP, Peng D, Su L, Xiao B and Zhang ZS: Curcumin protects intestinal mucosal barrier function of rat enteritis via activation of MKP-1 and attenuation of p38 and NF-kB Activation. PloS ONE 2010; 5(9):12969.
Cuzzocrea S, Chatterjee PK, Mazzon E, Dugo L, De Sarro A, Van de Loo FA, Caputi AP and Thiemermann C: Role of induced nitric oxide in the initiation of the inflammatory response after post-ischemic injury. Shock 2002; 18: 169-176.
Vermeiren GL, Claeys MJ, Van Bockstaele D, Grobben B, Slegers H, Bossaert L and Jorens PG: Re-perfusion injury after focal myocardial ischemia: Polymorphonuclear leukocyte activation and its clinical implications. Resuscitation 2002; 45: 35-61.
Grootjans J, Lenaerts K, Derikx JP, Matthijsen RA, de Bruine AP and van Bijnen AA. et al.: Human intestinal ischemia-reperfusion-induced inflammation characterized: experiences from a new translational model. Am J Pathol. 2010; 176(5):2283-91.
Ma NS: Changes in malondialdehyde contents in serum and tissues after ischemia and reperfusion of the bowel in dogs. Zhonghua Zheng Xing Shao Shang Wai Ke Za Zhi 1992; 8:127-129.
Lin ZL, Yu WK, Tan SJ, Duan KP, Dong Y., Bai XW, Xu L, and Li N: Protective effects of terminal ileostomy against bacterial translocation in a rat model of Intestinal ischemia/reperfusion injury. World J. Gastroenterol. 2014; 20:17905-17913.
Ocak T, Duran A, Ozyalvaçli G, Ocak Z, Terzi EH, Tosuns M and Erdem K: Protective effects of montelukast and Hypericum perforatum against intestinal ischemia-reperfusion injury in hamsters, Turk J. Med. Sci.2014; 44:381-386
Boybeyi O, Gunal YD, Atasoy P, Kısa U and Aslan MK: The effect of colchicines and low-dose methotrexate on intestinal ischemia/reperfusion injury in an experimental model, J. Pediatr. Surg. 2014; 49:1471-1474.
Tuboly E, Futakuchi M, a Varga G, Erces D, Tokes T, Meszaros A, Kaszaki J, Suzui M, Imai M, Okada A, Okada N, Boros M and Okada H: C5a inhibitor protects against ischemia/reperfusion injury in rat small intestine. Microbiol Immunol. 2016; 60: 35 – 46.
Carden DL and Granger DN: Pathophysiology of ischaemia-reperfusion injury. J Pathol. 2000; 190:255-266.
Park PO, Haglund U, Bulkley GB, and Falt K: The sequence of development of intestinal tissue injury after strangulation ischemia and reperfusion. Surgery 1990; 107:574-580.
Hacioglu A, Algin C, Pasaoglu O, Pasaoglu E and Kanbak G: Protective effect of leptin against ischemia – reperfusion injury in rat small intestine. BMC Gastroenterol. 2005; 5(37):1-7.
Kuo S, Liu Y, Tsai C and Sheen-Chen S: Ischemic preconditioning in hepatic ischemic – reperfusion injury. Formosan J. Surg. 2016; 49: 169 – 73.
Abd Allaha ESH, Ahmeda MA and Abdel Mola AF: Comparative study of the effect of verapamil and vitamin D on iron overload-induced oxidative stress and cardiac structural changes in adult male ratsPathophysiol. 2014; 21: 293 – 300.
Xu DZ, Lu Q, Kubicka R and Deitch EA: The effect of hypoxia/reoxygenation on the cellular function of intestinal epithelial cells. J. Trauma Fe 1999; 46(2):280-285.
Brower GL, Levick SP and Janicki JS.: Inhibition of matrix metalloproteinase activity by ACE inhibitors prevents left ventricular remodeling in a rat model of heart failure. Am. J. Physiol. Heart Circ. Physiol. 2007; 292: 3057 – 64.
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