Skip to main content
Top
Published in: Digestive Diseases and Sciences 8/2016

01-08-2016 | Original Article

Co-lyophilized Aspirin with Trehalose Causes Less Injury to Human Gastric Cells and Gastric Mucosa of Rats

Authors: Lee-Shuan Lin, Yuko Kayasuga-Kariya, Shugo Nakamura, Nobuyuki Shimohata, Takamasa Sakai, Ayano Fujisawa, Yuki Akagi, Shigeki Suzuki, Ung-il Chung, Nobuo Sasaki, Manabu Mochizuki

Published in: Digestive Diseases and Sciences | Issue 8/2016

Login to get access

Abstract

Background

Aspirin is one of the most popular NSAIDs worldwide because of its anti-inflammatory and anticoagulant effects, and however, gastrointestinal injury remains a major complication. We previously reported co-lyophilized aspirin/trehalose (Lyo A/T) decreased the aspirin-induced gastric lesions in dogs.

Aim

This study investigated the mechanism of gastroprotective effects of trehalose in vitro and in vivo.

Methods

The apoptotic assays were performed in a human gastric carcinoma cell line, which was treated with aspirin, mixed aspirin/trehalose (Mix A/T) or Lyo A/T. Gastric ulcer severity was examined after oral administration of drugs in rats. In addition, the mucosal tissue apoptotic status in drug-treated rats was evaluated. Molecular dynamics simulations and laser Raman spectroscopy were performed in order to examine the molecular properties of Lyo A/T.

Results

DNA fragmentation was detected in AGS cells that were treated with aspirin and Mix A/T, but not in the Lyo A/T-treated cells. There were fewer apoptotic cells in the Lyo A/T-treated cells than in the other cells. Gastric injury was reduced in rats that received oral Lyo A/T compared with the others, while PGE2 synthesis was equally decreased in all groups. TUNEL assay and immunohistochemistry of cleaved caspase-3 in the mucosal tissues also revealed that Lyo A/T treatment induced less apoptosis than the others. The Lyo A/T spectrum showed clear differences in several Raman bands compared with that of Mix A/T.

Conclusions

Our data showed that co-lyophilization of aspirin with trehalose reduced gastric injury, potentially through suppression of aspirin-induced mucosal cell apoptosis while retaining its anti-inflammatory effects.
Literature
2.
go back to reference Vane JR. Inhibition of prostaglandin synthesis as a mechanism of action for aspirin-like drugs. Nat New Biol. 1971;231:232–235.CrossRefPubMed Vane JR. Inhibition of prostaglandin synthesis as a mechanism of action for aspirin-like drugs. Nat New Biol. 1971;231:232–235.CrossRefPubMed
3.
go back to reference Wallace JL, Keenan CM, Granger DN. Gastric ulceration induced by nonsteroidal anti-inflammatory drugs is a neutrophil-dependent process. Am J Physiol. 1990;259:G462–G467.PubMed Wallace JL, Keenan CM, Granger DN. Gastric ulceration induced by nonsteroidal anti-inflammatory drugs is a neutrophil-dependent process. Am J Physiol. 1990;259:G462–G467.PubMed
4.
go back to reference Yoshida N, Yoshikawa T, Nakamura Y, et al. Role of neutrophil-mediated inflammation in aspirin-induced gastric mucosal injury. Dig Dis Sci. 1995;40:2300–2304.CrossRefPubMed Yoshida N, Yoshikawa T, Nakamura Y, et al. Role of neutrophil-mediated inflammation in aspirin-induced gastric mucosal injury. Dig Dis Sci. 1995;40:2300–2304.CrossRefPubMed
5.
go back to reference Kitahora T, Guth PH. Effect of aspirin plus hydrochloric acid on the gastric mucosal microcirculation. Gastroenterology. 1987;93:810–817.CrossRefPubMed Kitahora T, Guth PH. Effect of aspirin plus hydrochloric acid on the gastric mucosal microcirculation. Gastroenterology. 1987;93:810–817.CrossRefPubMed
6.
go back to reference Tanaka K, Tomisato W, Hoshino T, et al. Involvement of intracellular Ca2+ levels in nonsteroidal anti-inflammatory drug-induced apoptosis. J Biol Chem. 2005;280:31059–31067.CrossRefPubMed Tanaka K, Tomisato W, Hoshino T, et al. Involvement of intracellular Ca2+ levels in nonsteroidal anti-inflammatory drug-induced apoptosis. J Biol Chem. 2005;280:31059–31067.CrossRefPubMed
7.
go back to reference Tomisato W, Tsutsumi S, Hoshino T, et al. Role of direct cytotoxic effects of NSAIDs in the induction of gastric lesions. Biochem Pharmacol. 2004;67:575–585.CrossRefPubMed Tomisato W, Tsutsumi S, Hoshino T, et al. Role of direct cytotoxic effects of NSAIDs in the induction of gastric lesions. Biochem Pharmacol. 2004;67:575–585.CrossRefPubMed
8.
go back to reference Fiorucci S, Antonelli E, Santucci L, et al. Gastrointestinal safety of nitric oxide-derived aspirin is related to inhibition of ICE-like cysteine proteases in rats. Gastroenterology. 1999;116:1089–1106.CrossRefPubMed Fiorucci S, Antonelli E, Santucci L, et al. Gastrointestinal safety of nitric oxide-derived aspirin is related to inhibition of ICE-like cysteine proteases in rats. Gastroenterology. 1999;116:1089–1106.CrossRefPubMed
9.
go back to reference Fiorucci S, Santucci L, Antonelli E, et al. NO-aspirin protects from T cell-mediated liver injury by inhibiting caspase-dependent processing of Th1-like cytokines. Gastroenterology. 2000;118:404–421.CrossRefPubMed Fiorucci S, Santucci L, Antonelli E, et al. NO-aspirin protects from T cell-mediated liver injury by inhibiting caspase-dependent processing of Th1-like cytokines. Gastroenterology. 2000;118:404–421.CrossRefPubMed
10.
11.
go back to reference Lane ME, Kim MJ. Assessment and prevention of gastrointestinal toxicity of non-steroidal anti-inflammatory drugs. J Pharm Pharmacol. 2006;58:1295–1304.CrossRefPubMed Lane ME, Kim MJ. Assessment and prevention of gastrointestinal toxicity of non-steroidal anti-inflammatory drugs. J Pharm Pharmacol. 2006;58:1295–1304.CrossRefPubMed
12.
go back to reference Lim YJ, Lee JS, Ku YS, Hahm KB. Rescue strategies against non-steroidal anti-inflammatory drug-induced gastroduodenal damage. J Gastroenterol Hepatol. 2009;24:1169–1178.CrossRefPubMed Lim YJ, Lee JS, Ku YS, Hahm KB. Rescue strategies against non-steroidal anti-inflammatory drug-induced gastroduodenal damage. J Gastroenterol Hepatol. 2009;24:1169–1178.CrossRefPubMed
13.
15.
go back to reference Elbein AD. The metabolism of alpha, alpha-trehalose. Adv Carbohydr Chem Biochem. 1974;30:227–256.CrossRefPubMed Elbein AD. The metabolism of alpha, alpha-trehalose. Adv Carbohydr Chem Biochem. 1974;30:227–256.CrossRefPubMed
16.
go back to reference Benaroudj N, Lee DH, Goldberg AL. Trehalose accumulation during cellular stress protects cells and cellular proteins from damage by oxygen radicals. J Biol Chem. 2001;276:24261–24267.CrossRefPubMed Benaroudj N, Lee DH, Goldberg AL. Trehalose accumulation during cellular stress protects cells and cellular proteins from damage by oxygen radicals. J Biol Chem. 2001;276:24261–24267.CrossRefPubMed
17.
go back to reference Chen Q, Haddad GG. Role of trehalose phosphate synthase and trehalose during hypoxia: from flies to mammals. J Exp Biol. 2004;207:3125–3129.CrossRefPubMed Chen Q, Haddad GG. Role of trehalose phosphate synthase and trehalose during hypoxia: from flies to mammals. J Exp Biol. 2004;207:3125–3129.CrossRefPubMed
18.
go back to reference Echigo R, Shimohata N, Karatsu K, et al. Trehalose treatment suppresses inflammation, oxidative stress, and vasospasm induced by experimental subarachnoid hemorrhage. J Transl Med. 2012;10:80.CrossRefPubMedPubMedCentral Echigo R, Shimohata N, Karatsu K, et al. Trehalose treatment suppresses inflammation, oxidative stress, and vasospasm induced by experimental subarachnoid hemorrhage. J Transl Med. 2012;10:80.CrossRefPubMedPubMedCentral
19.
go back to reference Taya K, Hirose K, Hamada S. Trehalose inhibits inflammatory cytokine production by protecting IkappaB-alpha reduction in mouse peritoneal macrophages. Arch Oral Biol. 2009;54:749–756.CrossRefPubMed Taya K, Hirose K, Hamada S. Trehalose inhibits inflammatory cytokine production by protecting IkappaB-alpha reduction in mouse peritoneal macrophages. Arch Oral Biol. 2009;54:749–756.CrossRefPubMed
20.
go back to reference Minutoli L, Altavilla D, Bitto A, et al. The disaccharide trehalose inhibits proinflammatory phenotype activation in macrophages and prevents mortality in experimental septic shock. Shock. 2007;27:91–96.CrossRefPubMed Minutoli L, Altavilla D, Bitto A, et al. The disaccharide trehalose inhibits proinflammatory phenotype activation in macrophages and prevents mortality in experimental septic shock. Shock. 2007;27:91–96.CrossRefPubMed
21.
go back to reference Sasnoor LM, Kale VP, Limaye LS. Prevention of apoptosis as a possible mechanism behind improved cryoprotection of hematopoietic cells by catalase and trehalose. Transplantation. 2005;80:1251–1260.CrossRefPubMed Sasnoor LM, Kale VP, Limaye LS. Prevention of apoptosis as a possible mechanism behind improved cryoprotection of hematopoietic cells by catalase and trehalose. Transplantation. 2005;80:1251–1260.CrossRefPubMed
22.
go back to reference Chen W, Zhang X, Liu M, et al. Trehalose protects against ocular surface disorders in experimental murine dry eye through suppression of apoptosis. Exp Eye Res. 2009;89:311–318.CrossRefPubMed Chen W, Zhang X, Liu M, et al. Trehalose protects against ocular surface disorders in experimental murine dry eye through suppression of apoptosis. Exp Eye Res. 2009;89:311–318.CrossRefPubMed
23.
go back to reference Mori Y, Yano F, Shimohata N, Suzuki S, Chung UI, Takato T. Trehalose inhibits oral dryness by protecting the cell membrane. Int J Oral Maxillofac Surg. 2010;39:916–921.CrossRefPubMed Mori Y, Yano F, Shimohata N, Suzuki S, Chung UI, Takato T. Trehalose inhibits oral dryness by protecting the cell membrane. Int J Oral Maxillofac Surg. 2010;39:916–921.CrossRefPubMed
24.
go back to reference Fujino H, Lee S, Suzuki S, et al. Trehalose may prevent postsurgical adhesions in a rabbit model of hysterotomy. J Vet Med Sci. 2011;73:931–935.CrossRefPubMed Fujino H, Lee S, Suzuki S, et al. Trehalose may prevent postsurgical adhesions in a rabbit model of hysterotomy. J Vet Med Sci. 2011;73:931–935.CrossRefPubMed
25.
go back to reference Lin LS, Kayasuga Y, Shimohata N, et al. Lyophilized aspirin with trehalose may decrease the incidence of gastric injuries in healthy dogs. J Vet Med Sci. 2012;74:1511–1516.CrossRefPubMed Lin LS, Kayasuga Y, Shimohata N, et al. Lyophilized aspirin with trehalose may decrease the incidence of gastric injuries in healthy dogs. J Vet Med Sci. 2012;74:1511–1516.CrossRefPubMed
26.
go back to reference Kayasuga-Kariya Y, Iwanaga S, Fujisawa A, et al. Dermal cell damage induced by topical application of non-steroidal anti-inflammatory drugs is suppressed by trehalose co-lyophilization in ex vivo analysis. J Vet Med Sci. 2013;75:1619–1622.CrossRefPubMedPubMedCentral Kayasuga-Kariya Y, Iwanaga S, Fujisawa A, et al. Dermal cell damage induced by topical application of non-steroidal anti-inflammatory drugs is suppressed by trehalose co-lyophilization in ex vivo analysis. J Vet Med Sci. 2013;75:1619–1622.CrossRefPubMedPubMedCentral
27.
go back to reference Hess B, Kutzner C, van der Spoel D, Lindahl E. GROMACS 4: algorithms for highly efficient, load-balanced, and scalable molecular simulation. J Chem Theory Comput. 2008;4:435–447.CrossRefPubMed Hess B, Kutzner C, van der Spoel D, Lindahl E. GROMACS 4: algorithms for highly efficient, load-balanced, and scalable molecular simulation. J Chem Theory Comput. 2008;4:435–447.CrossRefPubMed
28.
29.
go back to reference Wang J, Wang W, Kollman PA, Case DA. Automatic atom type and bond type perception in molecular mechanical calculations. J Mol Graph Model. 2006;25:247–260.CrossRefPubMed Wang J, Wang W, Kollman PA, Case DA. Automatic atom type and bond type perception in molecular mechanical calculations. J Mol Graph Model. 2006;25:247–260.CrossRefPubMed
30.
go back to reference Lerbret A, Bordat P, Affouard F, Descamps M, Migliardo F. How homogeneous are the trehalose, maltose, and sucrose water solutions? An insight from molecular dynamics simulations. J Phys Chem B. 2005;109:11046–11057.CrossRefPubMed Lerbret A, Bordat P, Affouard F, Descamps M, Migliardo F. How homogeneous are the trehalose, maltose, and sucrose water solutions? An insight from molecular dynamics simulations. J Phys Chem B. 2005;109:11046–11057.CrossRefPubMed
31.
go back to reference Sapir L, Harries D. Linking trehalose self-association with binary aqueous solution equation of state. J Phys Chem B. 2011;115:624–634.CrossRefPubMed Sapir L, Harries D. Linking trehalose self-association with binary aqueous solution equation of state. J Phys Chem B. 2011;115:624–634.CrossRefPubMed
32.
go back to reference Hall AJ, Tripp M, Howell T, Darland G, Bland JS, Babish JG. Gastric mucosal cell model for estimating relative gastrointestinal toxicity of non-steroidal anti-inflammatory drugs. Prostaglandins Leukot Essent Fatty Acids. 2006;75:9–17.CrossRefPubMed Hall AJ, Tripp M, Howell T, Darland G, Bland JS, Babish JG. Gastric mucosal cell model for estimating relative gastrointestinal toxicity of non-steroidal anti-inflammatory drugs. Prostaglandins Leukot Essent Fatty Acids. 2006;75:9–17.CrossRefPubMed
33.
go back to reference Burgoyne LA, Hewish DR, Mobbs J. Mammalian chromatin substructure studies with the calcium-magnesium endonuclease and two-dimensional polyacrylamide-gel electrophoresis. Biochem J. 1974;143:67–72.CrossRefPubMedPubMedCentral Burgoyne LA, Hewish DR, Mobbs J. Mammalian chromatin substructure studies with the calcium-magnesium endonuclease and two-dimensional polyacrylamide-gel electrophoresis. Biochem J. 1974;143:67–72.CrossRefPubMedPubMedCentral
34.
go back to reference Oberhammer F, Wilson JW, Dive C, et al. Apoptotic death in epithelial cells: cleavage of DNA to 300 and/or 50 kb fragments prior to or in the absence of internucleosomal fragmentation. EMBO J. 1993;12:3679–3684.PubMedPubMedCentral Oberhammer F, Wilson JW, Dive C, et al. Apoptotic death in epithelial cells: cleavage of DNA to 300 and/or 50 kb fragments prior to or in the absence of internucleosomal fragmentation. EMBO J. 1993;12:3679–3684.PubMedPubMedCentral
35.
go back to reference Power JJ, Dennis MS, Redlak MJ, Miller TA. Aspirin-induced mucosal cell death in human gastric cells: evidence supporting an apoptotic mechanism. Dig Dis Sci. 2004;49:1518–1525.CrossRefPubMed Power JJ, Dennis MS, Redlak MJ, Miller TA. Aspirin-induced mucosal cell death in human gastric cells: evidence supporting an apoptotic mechanism. Dig Dis Sci. 2004;49:1518–1525.CrossRefPubMed
36.
go back to reference Archana M, Bastian YT, et al. Various methods available for detection of apoptotic cells—a review. Indian J Cancer. 2013;50:274–283.CrossRefPubMed Archana M, Bastian YT, et al. Various methods available for detection of apoptotic cells—a review. Indian J Cancer. 2013;50:274–283.CrossRefPubMed
37.
go back to reference Hernández C, Barrachina MD, Vallecillo-Hernández J, et al. Aspirin-induced gastrointestinal damage is associated with an inhibition of epithelial cell autophagy. J Gastroenterol. 2015. doi:10.1007/s00535-015-1137-1.PubMed Hernández C, Barrachina MD, Vallecillo-Hernández J, et al. Aspirin-induced gastrointestinal damage is associated with an inhibition of epithelial cell autophagy. J Gastroenterol. 2015. doi:10.​1007/​s00535-015-1137-1.PubMed
38.
go back to reference Sarkar S, Davies JE, Huang A, et al. Trehalose, a novel mTOR-independent autophagy enhancer, accelerates the clearance of mutant huntingtin and & α-synuclein. J Biol Chem. 2007;282:5641–5652.CrossRefPubMed Sarkar S, Davies JE, Huang A, et al. Trehalose, a novel mTOR-independent autophagy enhancer, accelerates the clearance of mutant huntingtin and & α-synuclein. J Biol Chem. 2007;282:5641–5652.CrossRefPubMed
39.
go back to reference Taylor LS, Zografi G. Sugar-polymer hydrogen bond interactions in lyophilized amorphous mixtures. J Pharm Sci. 1998;87:1615–1621.CrossRefPubMed Taylor LS, Zografi G. Sugar-polymer hydrogen bond interactions in lyophilized amorphous mixtures. J Pharm Sci. 1998;87:1615–1621.CrossRefPubMed
40.
go back to reference Davenport HW. Salicylate damage to the gastric mucosal barrier. N Engl J Med. 1967;276:1307–1312.CrossRefPubMed Davenport HW. Salicylate damage to the gastric mucosal barrier. N Engl J Med. 1967;276:1307–1312.CrossRefPubMed
41.
go back to reference Matsui H, Shimokawa O, Kaneko T, et al. The pathophysiology of non-steroidal anti-inflammatory drug (NSAID)-induced mucosal injuries in stomach and small intestine. J Clin Biochem Nutr. 2011;48:107–111.CrossRefPubMedPubMedCentral Matsui H, Shimokawa O, Kaneko T, et al. The pathophysiology of non-steroidal anti-inflammatory drug (NSAID)-induced mucosal injuries in stomach and small intestine. J Clin Biochem Nutr. 2011;48:107–111.CrossRefPubMedPubMedCentral
Metadata
Title
Co-lyophilized Aspirin with Trehalose Causes Less Injury to Human Gastric Cells and Gastric Mucosa of Rats
Authors
Lee-Shuan Lin
Yuko Kayasuga-Kariya
Shugo Nakamura
Nobuyuki Shimohata
Takamasa Sakai
Ayano Fujisawa
Yuki Akagi
Shigeki Suzuki
Ung-il Chung
Nobuo Sasaki
Manabu Mochizuki
Publication date
01-08-2016
Publisher
Springer US
Published in
Digestive Diseases and Sciences / Issue 8/2016
Print ISSN: 0163-2116
Electronic ISSN: 1573-2568
DOI
https://doi.org/10.1007/s10620-016-4209-z

Other articles of this Issue 8/2016

Digestive Diseases and Sciences 8/2016 Go to the issue