Culture Supernatants of Lactobacillus acidophilus and Bifidobacterium adolescentis Repress Ileal Ulcer Formation in Rats Treated with a Nonsteroidal Antiinflammatory Drug by Suppressing Unbalanced Growth of Aerobic Bacteria and Lipid Peroxidation

Takemi Kinouchi; Keiko Kataoka; Shan Ruo Bing; Haruyuki Nakayama; Motoo Uejima; Kazuyuki Shimono; Tomomi Kuwahara; Shigeru Akimoto; Isao Hiraoka; Yoshinari Ohnishi

Culture Supernatants of Lactobacillus acidophilus and Bifidobacterium adolescentis Repress Ileal Ulcer Formation in Rats Treated with a Nonsteroidal Antiinflammatory Drug by Suppressing Unbalanced Growth of Aerobic Bacteria and Lipid Peroxidation

Takemi Kinouchi, Keiko Kataoka, Shan Ruo Bing, Haruyuki Nakayama, Motoo Uejima, Kazuyuki Shimono, Tomomi Kuwahara, Shigeru Akimoto, Isao Hiraoka, Yoshinari Ohnishi

Author information

Takemi Kinouchi
Depament of Bacteriology, School of Medicine, The University of Tokushima
Keiko Kataoka
Depament of Bacteriology, School of Medicine, The University of Tokushima
Shan Ruo Bing
Depament of Bacteriology, School of Medicine, The University of Tokushima
Haruyuki Nakayama
Depament of Bacteriology, School of Medicine, The University of Tokushima
Motoo Uejima
Depament of Bacteriology, School of Medicine, The University of Tokushima
Naruto Research Institute, Otsuka Pharmaceutical Factory, Inc.
Kazuyuki Shimono
Depament of Bacteriology, School of Medicine, The University of Tokushima
Naruto Research Institute, Otsuka Pharmaceutical Factory, Inc.
Tomomi Kuwahara
Depament of Bacteriology, School of Medicine, The University of Tokushima
Shigeru Akimoto
Depament of Bacteriology, School of Medicine, The University of Tokushima
Isao Hiraoka
Naruto Research Institute, Otsuka Pharmaceutical Factory, Inc.
Yoshinari Ohnishi
Depament of Bacteriology, School of Medicine, The University of Tokushima

Keywords: NSAID, Ileal ulcer, Lipid peroxidation, Ileal microflora, Lactobacillus acidophilus, Bifidobacterium adolescentis

JOURNAL FREE ACCESS

1998 Volume 42 Issue 5 Pages 347-355

Details

Published: May 20, 1998 Received: October 14, 1997 Available on J-STAGE: March 17, 2008 Accepted: February 02, 1998 Advance online publication: - Revised: -
Correction information
Date of correction: March 17, 2008 Reason for correction: - Correction: TITLE Details: Wrong : Culture Supernatants of Lactobacillus acidophilus and Bifidobacterium adolescentis Repress Ileal Ulcer Formation in Rats Treated with a Nonsteroidal Antiinflammatory Drug by Suppressing Unbalanced Growth of Aerobic Bacteria and Lipid Peroxidation
Right : Culture Supernatants of Lactobacillus acidophilus and Bifidobacterium adolescentis Repress Ileal Ulcer Formation in Rats Treated with a Nonsteroidal Antiinflammatory Drug by Suppressing Unbalanced Growth of Aerobic Bacteria and Lipid Peroxidation

Date of correction: March 17, 2008 Reason for correction: - Correction: ABSTRACT Details: Wrong : A nonsteroidal antiinflammatory drug, 5-bromo-2-(4-fluorophenyl)-3-(4-methylsulfonylphenyl) thiophene (BFMeT), induced ileal ulcers in rats after oral administration, while no ulcers were observed after subcutaneous injection. The ileal ulcer formation in BFMeT-treated rats was examined to correlate the administration of cultures of Lactobacillus acidophilus or Bifidobacterium adolescentis with intestinal bacteria in the ileal contents and lipid peroxidation of the small intestinal mucosa. Ileal ulcers were observed in more than 85% of the rats treated with BFMeT at a dose of 1, 000mg/kg when they were given tap water as drinking water. The incidence of ulcer formation was repressed by giving culture supernatants of L. acidophilus or B. adolescentis as drinking water, but not by giving the cell suspension as drinking water. Gram staining of the ileal contents of normal rats revealed that 97% of the stained bacteria were Gram-positive rods and only 1.5% were Gram-negative rods. The percentage of Gram-negative rods 72hr after BFMeT administration was 49.8% and increased over 30-fold in BFMeT-treated rats. However, the percentage of Gram-negative rods was 9.7% or 16%, respectively, in rats taking culture supernatants of L. acidophilus or B. adolescentis. In addition, thiobarbituric acid-reactive substances in the ileal mucosa increased significantly in the rats given tap water for 72hr after BFMeT treatment, but not in rats given the culture supernatants of L. acidophilus or B. adolescentis. Since BFMeT induced an unbalanced intestinal microflora, the effect of antibiotic treatment on ulcer formation in rats was examined. The magnitude of the ulcer formation in the antibiotic-treated rats was, in decreasing order, metronidazole>none> kanamycin>a mixture (bacitracin, neomycin and streptomycin). These results suggest that the intestinal microflora plays an important role in ulcer formation and that a metabolite(s) of L. acidophilus and B. adolescentis inhibits ileal ulcer formation by repressing changes in the intestinal microflora and lipid peroxidation in BFMeT-treated rats.
Right : A nonsteroidal antiinflammatory drug, 5-bromo-2- (4-fluoropheny1) -3- (4-methylsulfonylphenyl) thiophene (BFMeT), induced ileal ulcers in rats after oral administration, while no ulcers were observed after subcutaneous injection. The ileal ulcer formation in BFMeT-treated rats was examined to correlate the administration of cultures of Lactobacillus acidophilus or Bifidobacterium adolescentis with intestinal bacteria in the ileal contents and lipid peroxidation of the small intestinal mucosa. Ileal ulcers were observed in more than 85% of the rats treated with BFMeT at a dose of 1, 000 mg/kg when they were given tap water as drinking water. The incidence of ulcer formation was repressed by giving culture supernatants of L. acidophilus or B. adolescentis as drinking water, but not by giving the cell suspension as drinking water. Gram staining of the ileal contents of normal rats revealed that 97 % of the stained bacteria were Gram-pos itive rods and only 1.5% were Gram-negative rods. The percentage of Gram-negative rods 72 hr after BFMeT administration was 49.8% and increased over 30-fold in BFMeT-treated rats. However, the percentage of Gram-negative rods was 9.7% or 16%, respectively, in rats taking culture supernatants of L. acidophilus or B. adolescentis. In addition, thiobarbituric acid-reactive substances in the ileal mucosa increased significantly in the rats given tap water for 72 hr after BFMeT treatment, but not in rats given the culture supernatants of L. acidophilus or B. adolescentis. Since BFMeT induced an unbalanced intestinal microflora, the effect of antibiotic treatment on ulcer formation in rats was examined. The magnitude of the ulcer formation in the antibiotic-treated rats was, in decreasing order, metronidazole > none > kanamycin > a mixture (bacitracin, neomycin and streptomycin). These results suggest that the intestinal microflora plays an important role in ulcer formation and that a metabolite (s) of L. acidophilus and B. adolescentis inhibits ileal ulcer formation by repressing changes in the intestinal microflora and lipid peroxi dation in BFMeT-treated rats.

Date of correction: March 17, 2008 Reason for correction: - Correction: DTPUB Details: Right : 19980520

Date of correction: March 17, 2008 Reason for correction: - Correction: AFFILIATION Details: Wrong :
1) Department of Bacteriology, School of Medicine, The University of Tokushima
2) Naruto Research Institute, Otsuka Pharmaceutical Factory, Inc.

Right :
1) Depament of Bacteriology, School of Medicine, The University of Tokushima
2) Naruto Research Institute, Otsuka Pharmaceutical Factory, Inc.

Date of correction: March 17, 2008 Reason for correction: - Correction: KEYWORD Details: Wrong : NSAID, Ileal ulcer, Lipid peroxidation, Ileal microflora, Lactobacillus acidophilus, Bifidobacterium adolescentis
Right : NSAID, Ileal ulcer, Lipid peroxidation, Ileal microflora, Lactobacillus acidophilus, Bifidobacterium adolescentis

Date of correction: March 17, 2008 Reason for correction: - Correction: CITATION Details: Wrong : 1) Allison, M.C., Howatson, A.G., Torrance, C.J., Lee, F.D., and Russell, R.I. 1992. Gastrointestinal damage associated with the use of nonsteroidal antiinflammatory drugs. N. Engl. J. Med. 327: 749-754.
2) Beck, W.S., Schneider, H.T., Dietzel, K., Nuernberg, B., and Brune, K. 1990. Gastrointestinal ulcerations induced by anti-inflammatory drugs in rats. Arch. Toxicol. 64: 210-217.
3) Bjarnason, I., Hayllar, J., Macpherson, A.J., and Russell, A.S. 1993. Side effects of nonsteroidal anti-inflammatory drugs on the small and large intestine in humans. Gastroenterology 104: 1832-1847.
4) Bjarnason, I., Zanelli, G., Smith, T., Prouse, P., Williams, P., Smethurst, P., Delacey, G., Gumpel, M.J., and Levi, A.J. 1987. Nonsteroidal antiinflammatory drug-induced intestinal inflammation in humans. Gastroenterology 93: 480-489.
5) Deakin, M. 1988. Small bowel perforation associated with an excessive dose of slow release diclofenac sodium. Br. Med. J.297: 488-489.
6) Dohn, D.R., Leininger, J.R., Lash, L.H., Quebbemann, A.J., and Anders, M.W. 1985. Nephrotoxicity of S-(2-chloro-1,1,2-trifluoroethyl)glutathione and S-(2-chloro-1,1,2-trifluoroethyl)-L-cysteine, the glutathione and cysteine conjugates of chlorotrifluoroethene. J. Pharmacol. Exp. Ther. 235: 851-857.
7) Duggan, D.E., Hooke, K.F., Noll, R.M., and Kwan, K.C. 1975. Enterohepatic circulation of indomethacin and its role in intestinal irritation. Biochem. Pharmacol. 25: 1749-1754.
8) Elfarra, A.A., Lash, L.H., and Anders, M.W. 1986. Metabolic activation and detoxication of nephrotoxic cysteine and homocysteine S-conjugates. Proc. Natl. Acad. Sci. U.S.A. 83: 2667-2671.
9) Freeman, B.A., and Crapo, J.D. 1982. Biology of disease: free radicals and tissue injury. Lab. Invest. 47: 412-425.
10) Gans, K.R., Galbraith, W., Roman, R.J., Haber, S.B., Kerr, J.S., Schmidt, W.K., Smith, C., Hewes, W.E., and Ackerman, N.R. 1990. Anti-inflammatory and safety profile of DuP 697, a novel orally effective prostaglandin synthesis inhibitor. J. Pharmacol. Exp. Ther. 254: 180-187.
11) Goldin, B.R. 1986. In situ bacterial metabolism and colon mutagens. Annu. Rev. Microbiol. 40: 367-393.
12) Hamilton, J.D., Dyer, N.H., Dawson, A.M., O'Grady, F.W., Vince, A., Fenton, J.C.B., and Mollin, D.L. 1970. Assessment and significance of bacterial overgrowth in the small bowel. Quart. J. Med. 39: 265-285.
13) Holst, O., Ulmer, A.J., Brade, H., Flad, H.-D., and Rietschel, E.T. 1996. Biochemistry and cell biology of bacterial endotoxins. FEMS Immunol. Med. Microbiol. 16: 83-104.
14) Kataoka, K., Kinouchi, T., Akimoto, S., and Ohnishi, Y. 1995. Bioactivation of cysteine conjugates of 1-nitropyrene oxides by cysteine conjugate β-lyase purified from Peptostreptococcus magnus. Appl. Environ. Microbiol. 61: 3781-3787.
15) Kent, T.H., Osborne, J.W., and Wende, C.M. 1968. Intestinal flora in whole-body and intestinal X-irradiated rats. Radiat. Res. 35: 635-651.
16) Kinouchi, T., Kataoka, K., Miyanishi, K., Akimoto, S., and Ohnishi, Y. 1993. Biological activities of the intestinal microflora in mice treated with antibiotics or untreated and the effects of the microflora on absorption and metabolic activation of orally administered glutathione conjugates of K-region epoxides of 1-nitropyrene. Carcinogenesis 14: 869-874.
17) Kinouchi, T., Nishifuji, K., and Ohnishi, Y. 1987. In vitro intestinal microflora-mediated metabolism of biliary metabolites from 1-nitropyrene-treated rats. Microbiol. Immunol. 31:1145-1159.
18) Kinouchi, T., Nishifuji, K., and Ohnishi, Y. 1990. Biliary excretion of glutathione conjugates of 4,5-epoxy-4,5-dihydro-1-nitropyrene and 9,10-epoxy-9,10-dihydro-1-nitropyrene in rats administered 1-nitropyrene orally and their further metabolism in the intestinal tract. Carcinogenesis 11: 1381-1387.
19) Kinouchi, T., and Ohnishi, Y. 1986. Metabolic activation of 1-nitropyrene and 1,6-dinitropyrene by nitroreductases from Bacteroides fragilis and distribution of nitroreductase activity in rats. Microbiol. Immunol. 30: 979-992.
20) Langman, M.J.S. 1989. Epidemiologic evidence on the association between peptic ulceration and antiinflammatory drug use. Gastroenterology 96: 640-646.
21) Langman, M.J.S., Morgan, L., and Worrall, A. 1985. Use of anti-inflammatory drugs by patients admitted with small or large bowel perforations and hemorrhage. Br. Med. J. 290: 347-349.
22) Lowry, O.H., Rosebrough, N.J., Farr, A.L., and Randall, R.J. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193: 265-275.
23) Mai, U.E.H., Perez-Perez, G.I., Wahl, L.M., Wahl, S.M., Blaser, M.J., and Smith, P.D. 1991. Soluble surface proteins from Helicobacter pylori activate monocytes/macrophages by lipopolysaccharide-independent mechanism. J. Clin. Invest. 87: 894-900.
24) Monks, T.J., Anders, M.W., Dekant, W., Stevens, J.L., Lau, S.S., and Van Bladeren, P.J. 1990. Glutathione conjugate mediated toxicities. Toxicol. Appl. Pharmacol. 106: 1-19.
25) Nagaoka, M., Hashimoto, S., Watanabe, T., Yokokura, T., and Mori, Y. 1994. Anti-ulcer effects of lactic acid bacteria and their cell wall polysaccharides. Biol. Pharm. Bull. 17: 1012-1017.
26) Nakayama, H., Kinouchi, T., Kataoka, K., Akimoto, S., Matsuda, Y., and Ohnishi, Y. 1997. Intestinal anaerobic bacteria hydrolyse sorivudine, producing the high blood concentration of 5-(E)-(2-bromovinyl)uracil that increases the level and toxicity of 5-fluorouracil. Pharmacogenetics 7: 35-43.
27) Ohkawa, H., Ohishi, N., and Yagi, K. 1979. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal. Biochem. 95: 351-358.
28) Parks, D.A., Bulkley, G.B., Granger, D.N., Hamilton, S.R., and McCord, J.M. 1982. Ischemic injury in the cat small intestine: role of superoxide radicals. Gastroenterology 82: 9-15.
29) Pemberton, R.E., and Strand, L.J. 1979. A review of upper-gastrointestinal effects of the newer nonsteroidal antiinflammatory agents. Dig. Dis. Sci. 24: 53-62.
30) Rainsford, K.D. 1982. An analysis of the gastro-intestinal side-effects of non-steroidal anti-inflammatory drugs, with particular reference to comparative studies in man and laboratory species. Rheumatol. Int. 2: 1-10.
31) Rainsford, K.D. 1989. Mechanisms of gastrointestinal toxicity of non-steroidal anti-inflammatory drugs. Scand. J. Gastroenterol. 24 (Suppl 163): 9-16.
32) Scheline, R R. 1973. Metabolism of foreign components by gastrointestinal microorganisms. Pharmacol. Rev. 25: 451-523.
33) Somasundaram, S., Hayllar, H., Rafi, S., Wrigglesworth, J.M., Macpherson, A.J.S., and Bjarnason, 1. 1995. The biochemical basis of non-steroidal anti-inflammatory drug-induced damage to the gastrointestinal tract: a review and hypothesis. Scand. J. Gastroenterol. 30: 289-299.
34) Sun, D.C.H., Roth, S.H., Mitchell, C.S., and Englund, D.W. 1974. Upper gastrointestinal disease in rheumatoid arthritis. Am. J. Dig. Dis. 19: 405-4 10.
35) Tannock, G.W., and Savage, D.C. 1974. Influences of dietary and environmental stress on microbial populations in the murine gastrointestinal tract. Infect. Immun. 9: 591-598.
36) Uejima, M., Kinouchi, T., Kataoka, K., Hiraoka, I., and Ohnishi, Y. 1996. Role of intestinal bacteria in ileal ulcer formation in rats treated with a nonsteroidal antiinflammatory drug. Microbiol. Immunol. 40: 553-560.
37) Wallace, J.L., Arfors, K.-E., and McKnight, G.W. 1991. A monoclonal antibody against the CD18 leukocyte adhesion molecule prevents indomethacin-induced gastric damage in the rabbit. Gastroenterology 100: 878-883.
38) Wallace, J.L., Keenan, C.M., and Granger, D.N. 1990. Gastric ulceration induced by nonsteroidal anti-inflammatory drugs is a neutrophil-dependent process. Am. J. Physiol. 259: G462-G467.
39) Yoshikawa, T., Naito, Y., Kishi, A., Tomii, T., Kaneko, T., Iinuma, S., Ichikawa, H., Yasuda, M., Takahashi, S., and Kondo, M. 1993. Role of active oxygen, lipid peroxidation, and antioxidants in the pathogenesis of gastric mucosal injury induced by indomethacin in rats. Gut 34: 732-737.

Right : 1) Allison, M.C., Howatson, A.G., Torrance, C.J., Lee, F.D., and Russell, R.I. 1992. Gastrointestinal damage associated with the use of nonsteroidal antiinflammatory drugs. N. Engl. J. Med. 327 : 749-754.
2) Beck, W.S., Schneider, H.T., Dietzel, K., Nuernberg, B., and Brune, K. 1990. Gastrointestinal ulcerations induced by anti-inflammatory drugs in rats. Arch. Toxicol. 64 : 210-217.
3) Bjarnason, I., Hayllar, J., Macpherson, A.J., and Russell, A.S. 1993. Side effects of nonsteroidal anti-inflammatory drugs on the small and large intestine in humans. Gastroenterology 104 : 1832-1847.
4) Bjarnason, I., Zanelli, G., Smith, T., Prouse, P., Williams, P., Smethurst, P., Delacey, G., Gumpel, M.J., and Levi, A.J. 1987. Nonsteroidal antiinflammatory drug-induced intestinalinflammation in humans. Gastroenterology 93 : 480-489.
5) Deakin, M. 1988. Small bowel perforation associated with an excessive dose of slow release diclofenac sodium. Br. Med. J. 297 : 488-489.
6) Dohn, D.R., Leininger, J.R., Lash, L.H., Quebbemann, A.J., and Anders, M.W. 1985. Nephrotoxicity of S- (2-chloro1, 1, 2-trifluoroethyl) glutathione and S- (2-chloro-1, 1, 2-trifluoroethyl) -L-cysteine, the glutathione and cysteine conjugates of chlorotrifluoroethene. J. Pharmacol. Exp. Ther. 235 : 851-857.
7) Duggan, D.E., Hooke, K.F., Noll, R.M., and Kwan, K.C. 1975. Enterohepatic circulation of indomethacin and its role in intestinal irritation. Biochem. Pharmacol. 25 : 1749-1754.
8) Elfarra, A.A., Lash, L.H., and Anders, M.W. 1986. Metabolic activation and detoxication of nephrotoxic cysteine and homocysteine S-conjugates. Proc. Natl. Acad. Sci. U.S.A. 83 : 2667-2671.
9) Freeman, B.A., and Crapo, J.D. 1982. Biology of disease : free radicals and tissue injury. Lab. Invest. 47 : 412-425.
10) Gans, K.R., Galbraith, W., Roman, R.J., Haber, S.B., Kerr, J.S., Schmidt, W.K., Smith, C., Hewes, WE., and Ackerman, N.R. 1990. Anti-inflammatory and safety profile of DuP 697, a novel orally effective prostaglandin synthesis inhibitor. J. Pharmacol. Exp. Ther. 254 : 180-187.
11) Goldin, B.R. 1986. In situ bacterial metabolism and colon mutagens. Annu. Rev. Microbiol. 40 : 367-393.
12) Hamilton, J.D., Dyer, N.H., Dawson, A.M., O'Grady, F.W., Vince, A., Fenton, J.C.B., and Mollin, D.L. 1970. Assessment and significance of bacterial overgrowth in the small bowel. Quart. J. Med. 39 : 265-285.
13) Holst, O., Ulmer, A.J., Brade, H., Flad, H.-D., and Rietschel, E.T. 1996. Biochemistry and cell biology of bacterial endotoxins. FEMS Immunol. Med. Microbiol. 16 : 83-104.
14) Kataoka, K., Kinouchi, T., Akimoto, S., and Ohnishi, Y. 1995. Bioactivation of cysteine conjugates of 1-nitropyrene oxides by cysteine conjugate β-lyase purified from Peptostreptococcus magnus. Appl. Environ. Microbiol. 61 : 3781-3787.
15) Kent, T.H., Osborne, J.W., and Wende, C.M. 1968. Intestinal flora in whole-body and intestinal X-irradiated rats. Radiat. Res. 35 : 635-651.
16) Kinouchi, T., Kataoka, K., Miyanishi, K., Akimoto, S., and Ohnishi, Y. 1993. Biological activities of the intestinal microflora in mice treated with antibiotics or untreated and the effects of the microflora on absorption and metabolic activation of orally administered glutathione conjugates of Kregion epoxides of 1-nitropyrene. Carcinogenesis 14 : 869-874.
17) Kinouchi, T., Nishifuji, K., and Ohnishi, Y. 1987. In vitro intestinal microflora-mediated metabolism of biliary metabolites from 1-nitropyrene-treated rats. Microbiol. Immunol. 31 : 1145-1159.
18) Kinouchi, T., Nishifuji, K., and Ohnishi, Y. 1990. Biliary excretion of glutathione conjugates of 4, 5-epoxy-4, 5-dihydro1-nitropyrene and 9, 10-epoxy-9, 10-dihydro-1-nitropyrene in rats administered 1-nitropyrene orally and their further metabolism in the intestinal tract. Carcinogenesis 11 : 1381-1387.
19) Kinouchi, T., and Ohnishi, Y. 1986. Metabolic activation of 1-nitropyrene and 1, 6-dinitropyrene by nitroreductases from Bacteroides fragilis and distribution of nitroreductase activity in rats. Microbiol. Immunol. 30 : 979-992.
20) Langman, M.J.S. 1989. Epidemiologic evidence on the association between peptic ulceration and antiinflammatory drug use. Gastroenterology 96 : 640-646.
21) Langman, M.J.S., Morgan, L., and Worrall, A. 1985. Use of anti-inflammatory drugs by patients admitted with small or large bowel perforations and hemorrhage. Br. Med. J. 290 : 347-349.
22) Lowry, O.H., Rosebrough, N.J., Farr, A.L., and Randall, R.J. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193 : 265-275.
23) Mai, U.E.H., Perez-Perez, G.I., Wahl, L.M., Wahl, S.M., Blaser, M.J., and Smith, P.D. 1991. Soluble surface proteins from Helicobacter pylori activate monocytes/macrophages by lipopolysaccharide-independent mechanism. J. Clin. Invest. 87 : 894-900.
24) Monks, T.J., Anders, M.W., Dekant, W., Stevens, J.L., Lau, S.S., and Van Bladeren, P.J. 1990. Glutathione conjugate mediated toxicities. Toxicol. Appl. Pharmacol. 106 : 1-19.
25) Nagaoka, M., Hashimoto, S., Watanabe, T., Yokokura, T., and Mori, Y. 1994. Anti-ulcer effects of lactic acid bacteria and their cell wall polysaccharides. Biol. Pharm. Bull. 17 : 1012-1017.
26) Nakayama, H., Kinouchi, T., Kataoka, K., Akimoto, S., Matsuda, Y., and Ohnishi, Y. 1997. Intestinal anaerobic bac teria hydrolyse sorivudine, producing the high blood concentration of 5- (E) - (2-bromovinyl) uracil that increases the level and toxicity of 5-fluorouracil. Pharmacogenetics 7 : 35-43.
27) Ohkawa, H., Ohishi, N., and Yagi, K. 1979. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal. Biochem. 95 : 351-358.
28) Parks, D.A., Bulkley, G.B., Granger, D.N., Hamilton, S.R., and McCord, J.M. 1982. Ischemic injury in the cat small intestine : role of superoxide radicals. Gastroenterology 82 : 9-15.
29) Pemberton, R.E., and Strand, L.J. 1979. A review of uppergastrointestinal effects of the newer nonsteroidal antiinflammatory agents. Dig. Dis. Sci. 24 : 53-62.
30) Rainsford, K.D. 1982. An analysis of the gastro-intestinal side-effects of non-steroidal anti-inflammatory drugs, with particular reference to comparative studies in man and laboratory species. Rheumatol. Int. 2 : 1-10.
31) Rainsford, K.D. 1989. Mechanisms of gastrointestinal toxicity of non-steroidal anti-inflammatory drugs. Scand. J. Gastroenterol. 24 (Suppl 163) : 9-16.
32) Scheline, R R. 1973. Metabolism of foreign components by gastrointestinal microorganisms. Pharmacol. Rev. 25 : 451-523.
33) Somasundaram, S., Hayllar, H., Rafi, S., Wrigglesworth, J.M., Macpherson, A.J.S., and Bjarnason, I. 1995. The biochemical basis of non-steroidal anti-inflammatory druginduced damage to the gastrointestinal tract : a review and hypothesis. Scand. J. Gastroenterol. 30 : 289-299.
34) Sun, D.C.H., Roth, S.H., Mitchell, C.S., and Englund, D.W. 1974. Upper gastrointestinal disease in rheumatoid arthritis. Am. J. Dig. Dis. 19 : 405-410.
35) Tannock, G.W., and Savage, D.C. 1974. Influences of dietary and environmental stress on microbial populations in the murine gastrointestinal tract. Infect. Immun. 9 : 591-598.
36) Uejima, M., Kinouchi, T., Kataoka, K., Hiraoka, I., and Ohnishi, Y. 1996. Role of intestinal bacteria in ileal ulcer formation in rats treated with a nonsteroidal antiinflammatory drug. Microbiol. Immunol. 40 : 553-560.
37) Wallace, J.L., Arfors, K.-E., and McKnight, G.W. 1991. A monoclonal antibody against the CD18 leukocyte adhesion molecule prevents indomethacin-induced gastric damage in the rabbit. Gastroenterology 100 : 878-883.
38) Wallace, J.L., Keenan, C.M., and Granger, D.N. 1990. Gastric ulceration induced by nonsteroidal anti-inflammatory drugs is a neutrophil-dependent process. Am. J. Physiol. 259 : G462-G467.
39) Yoshikawa, T., Naito, Y., Kishi, A., Tomii, T., Kaneko, T., Iinuma, S., Ichikawa, H., Yasuda, M., Takahashi, S., and Kondo, M. 1993. Role of active oxygen, lipid peroxidation, and antioxidants in the pathogenesis of gastric mucosal injury induced by indomethacin in rats. Gut 34 : 732-737.

Abstract

A nonsteroidal antiinflammatory drug, 5-bromo-2- (4-fluoropheny1) -3- (4-methylsulfonylphenyl) thiophene (BFMeT), induced ileal ulcers in rats after oral administration, while no ulcers were observed after subcutaneous injection. The ileal ulcer formation in BFMeT-treated rats was examined to correlate the administration of cultures of Lactobacillus acidophilus or Bifidobacterium adolescentis with intestinal bacteria in the ileal contents and lipid peroxidation of the small intestinal mucosa. Ileal ulcers were observed in more than 85% of the rats treated with BFMeT at a dose of 1, 000 mg/kg when they were given tap water as drinking water. The incidence of ulcer formation was repressed by giving culture supernatants of L. acidophilus or B. adolescentis as drinking water, but not by giving the cell suspension as drinking water. Gram staining of the ileal contents of normal rats revealed that 97 % of the stained bacteria were Gram-pos itive rods and only 1.5% were Gram-negative rods. The percentage of Gram-negative rods 72 hr after BFMeT administration was 49.8% and increased over 30-fold in BFMeT-treated rats. However, the percentage of Gram-negative rods was 9.7% or 16%, respectively, in rats taking culture supernatants of L. acidophilus or B. adolescentis. In addition, thiobarbituric acid-reactive substances in the ileal mucosa increased significantly in the rats given tap water for 72 hr after BFMeT treatment, but not in rats given the culture supernatants of L. acidophilus or B. adolescentis. Since BFMeT induced an unbalanced intestinal microflora, the effect of antibiotic treatment on ulcer formation in rats was examined. The magnitude of the ulcer formation in the antibiotic-treated rats was, in decreasing order, metronidazole > none > kanamycin > a mixture (bacitracin, neomycin and streptomycin). These results suggest that the intestinal microflora plays an important role in ulcer formation and that a metabolite (s) of L. acidophilus and B. adolescentis inhibits ileal ulcer formation by repressing changes in the intestinal microflora and lipid peroxi dation in BFMeT-treated rats.

Corresponding author

Correction information

Register with J-STAGE for free!