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Specific Preinduction of 60-kDa Heat Shock Protein (Chaperonin Homolog) by TRH Does Not Protect Colonic Mucosa Against Acetic Acid-Induced Lesion in Rats

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Abstract

In order to study the cytoprotective function of colonic heat shock proteins (HSPs) in vivo, the effect of specific preinduction of HSP60 by thyrotropin-releasing hormone (TRH) administration on the development of acetic acid-induced colonic mucosal lesion was investigated. Expression of 60-kDa, 72-kDa, and 90-kDa heat shock proteins (HSP60, HSP72, and HSP90, respectively) in rat colonic mucosa was investigated by western blot and immunohistochemical analyses before and after TRH administration. Following pretreatment with or without TRH administration, the rats received intrarectal infusion of 5% acetic acid. The colonic mucosal damage was macroscopically evaluated 24 hr after the intrarectal infusion of acetic acid. Expression of HSP60 was significantly increased by TRH administration in the colonic mucosa, whereas HSP72 and HSP90 did not increase. Immunohistochemical study also showed a significant increase in HSP60 in colonic mucosal cells, especially at the surface of the colonic mucosa after TRH administration. No histopathologic alteration was observed in the colonic mucosa after TRH administration. The colonic mucosal damage caused by intrarectal infusion of 5% acetic acid was not prevented by preinduction of HSP60. We demonstrated that specific preinduction of HSP60 by TRH administration did not show cytoprotective function in the colonic mucosa, although this protein plays a crucial role for cytoprotection in the pancreatic acinar cells. Our results indicate that the role of HSP60 may be different in each organ with respect to cytoprotection.

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REFERENCES

  1. Itoh H, Tashima Y: The stress (heat shock) proteins. Int J Biochem 23:1185–1191, 1991

    Google Scholar 

  2. Hightower LE: Cultured animal cells exposed to amino acid analogues or puromycin rapidly synthesis several polypeptides. J Cell Physiol 102:407–427, 1980

    Google Scholar 

  3. Tissieres A, Mictchell HK, Tracy VM: Protein synthesis in salivary glands of Drosophilia melanogaster: Relation to chromosome puffs. J Mol Biol 84:389–393, 1974

    Google Scholar 

  4. Nakamura K, Rokutan K, Marui N, Aoike A, Kawai K: Induction of heat shock proteins and their implication in protection against ethanol-induced damage in cultured guinea pig gastric mucosal cells. Gastroenterology 101:161–166, 1991

    Google Scholar 

  5. Zeniya A, Otaka M, Itoh H, Kuwabara T, Fujimori S, Otani S, Tashima Y, Masamune O: Induction and intracellular localization of a 72-kDa heat shock protein in rat gastric mucosa after water-immersion stress. J Gastroenterol 30:572–577, 1995

    Google Scholar 

  6. Otaka M, Itoh H, Kuwabara T, Zeniya A, Fujimori S, Otani S, Tashima Y, Masamune O: Induction of heat shock protein and prevention of cerulein-induced pancreatitis by waterimmersion stress in rats. Int J Biochem 26:805–811, 1994

    Google Scholar 

  7. Otaka M, Okuyama A, Otani S, Jin M, Itoh S, Itoh H, Iwabuchi A, Sasahara H, Odashima M, Tashima Y, Masamune O: Differential induction of HSP60 and HSP72 by different stress situations in rats. Correlation with cerulein-induced pancreatitis. Dig Dis Sci 42:1473–1479, 1997

    Google Scholar 

  8. Otani S, Otaka M, Jin M, Okuyama A, Itoh S, Iwabuchi A, Sasahara H, Itoh H, Tashima Y, Masamune O: Effect of preinduction of heat shock proteins on acetic acid-induced colitis in rats. Dig Dis Sci 42:833–846, 1997

    Google Scholar 

  9. Stojadinovic A, Kiang J, Smallridge R, Gallowy R, Shea-Donohue T: Induction of heat shock protein 72 protects against ischemia/reperfusion in rat small intestine. Gastroenterology 109:505–515, 1995

    Google Scholar 

  10. Sasahara H, Otaka M, Itoh S, Iwabuchi A, Odashima M, Wada I, Konishi N, Pacheco I, Tashima Y, Itoh H, Otani S, Masamune O: Effect of pre-induction of heat shock proteins on acetic acid-induced small intestinal lesion in rats. Dig Dis Sci 43:2117–2130, 1998

    Google Scholar 

  11. Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254, 1976

    Google Scholar 

  12. Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685, 1970

    Google Scholar 

  13. Towbin H, Staehelin T, Gordon J: Electrophoretic transfer of protein from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354, 1979

    Google Scholar 

  14. Otaka M, Itoh H, Kuwabara T, Zeniya A, Fujimori S, Otani S, Tashima Y, Masamune O: Induction of a 60-kDa heat shock protein in rat pancreas by water-immersion stress. Int J Biochem 25:1769–1773, 1993

    Google Scholar 

  15. Wakui H, Itoh H, Tashima Y, Kobayashi R, Nakamoto Y, Miura AB: Specific antibodies against the stress-inducible 72-kDa protein, a member of the heat-shock protein hsp72, in healthy human subjects. Int J Biochem 23:975–978, 1991

    Google Scholar 

  16. Itoh H, Toyoshima I, Mizunaka H, Kobayashi R, Tashima Y: Three-step purification method and characterization of the bovine brain 90-kDa heat shock protein. Arch Biochem Biophys 282:290–296, 1990

    Google Scholar 

  17. McLean IW, Nakane PK: Periodate-lysine-paraformaldehyde fixative: A new fixative for immunoelectron microscopy. J Histochem Cytochem 22:1077–1083, 1974

    Google Scholar 

  18. John PD, Margaret B: Acute radiation enteritis in rats: Bile salts and trypsin. Surgery 112:587–592, 1992

    Google Scholar 

  19. Yano S, Akahane M, Harada M: Role of gastric motility in development of stress-induced gastric lesions of rats. Jpn J Pharmacol 28:607–615, 1978

    Google Scholar 

  20. Bueno L, Gue M: Evidence of the involvement of corticotropin-releasing factor in the gastrointestinal disturbances induced by acoustic and cold stress in mice. Brain Res 441:1–4, 1992

    Google Scholar 

  21. Bueno L, Fioramonti J: Neurohormonal control of intestinal transit. Reprod Nutr Dev 34:513–525, 1994

    Google Scholar 

  22. Axelrod J, Reisine T: Stress hormones; Their interaction and regulation. Science 224:452–459, 1987

    Google Scholar 

  23. Dubois M, Pickar D, Cohen MR, Roth YF, Macnamara T, Bunney WE Jr: Surgical stress in humans is accompanied by an increase in plasma beta-endorphin immunoreactivity. Life Sci 29:1249–1254, 1981

    Google Scholar 

  24. Williams CL, Peterson JM, Villar RG, Burks TF: Corticotropin releasing factor directly mediates colonic responses to stress. Am J Physiol 253:G582-G586, 1987

    Google Scholar 

  25. Nakae T, Audhya T, Hollander CS: Evidence for a role of endogenous corticotropin-releasing factor in cold, ether, immobilization, and traumatic stress. Proc Natl Acad Sci 82:1245–1251, 1985

    Google Scholar 

  26. Hashimoto K, Suemaru S, Takao T, Sugawara M, Makino S, Ota Z: Corticotropin-releasing factor and pituitary adenocortical responses in chronically stressed rats. Regul Pept 23:117–126, 1988

    Google Scholar 

  27. Horita A, Carino MA: Centrally administered thyrotropin releasing hormone (TRH) stimulates colonic transit and diarrhea production by a vagally mediated serotonergic mechanism in the rabbit. J Pharmacol Exp Ther 222:367–371, 1982

    Google Scholar 

  28. Ahlman H, Dahlstrom A: Vagal mechanisms controlling serotonin release from the gastrointestinal tract and pyloric motor function. J Auton Nerv Syst 9:119–140, 1983

    Google Scholar 

  29. Horita A, Carino MA, Pae YS: Blockade of naloxone and naltrexone of the TRH-induced stimulation of colonic transit in the rabbit. Eur J Pharmacol 108:289–293, 1985

    Google Scholar 

  30. Sharma HS, Dey PK: Impairment of blood-brain barrier (BBB) in rats by immobilization stress: Role of serotonin (5-HT). Indian J Physiol 25:111–112, 1981

    Google Scholar 

  31. Koivusalo F, Leppaluoto J: Brain TRH immunoreactivity during various physiological and stress conditions in rat. Neuroendocrinology 29:231–236, 1979

    Google Scholar 

  32. Arancibia S, Tapia-Arancibia L, Assenmacher I, Astier H: Direct evidence of short-term cold-induced TRH release in the median eminence of unanesthetized rats. Neuroendocrinology 37:225–228, 1983

    Google Scholar 

  33. Kubek MJ, Sattin A: Effects of electronconvulsive shock on the content of thyrotropin-releasing hormone in rat brain. Life Sci 34:598–602, 1984

    Google Scholar 

  34. Horita A, Carino MA, Mori T: Morphine antagonizes TRH-induced intestinal transit and hyperserotonemia in rabbit. Euro J Pharmacol 145:55–59, 1988

    Google Scholar 

  35. Miyata K, Kamato T, Nishida A, Ito H, Yuki H, Yamano M, Tsutsumi R, Katsuyama Y, Honda K: Role of the serotonin 3 receptor in stress-induced defecation. J Pharmacol Exp Ther 261:297–303, 1992

    Google Scholar 

  36. Costa M, Furness JB, Cuello ACU, Verhostad AA, Steinbusch HW, Elde RP: Neurons with 5-hydroxytryptamine-like immunoreactivity in the enteric nervous system: Their visualization and reactions to drug treatment. Neuroscience 7:351–363, 1982

    Google Scholar 

  37. King FD, Sanger GJ: 5-HT3 receptor antagonist. Drug Future 14:875–889, 1989

    Google Scholar 

  38. Ikeda T, Kitajima M: The study of biogenic amines following thermal injury with special reference to histamine and serotonin. Prog Med 7:419–424, 1987

    Google Scholar 

  39. Kuwabara T, Otaka M, Itoh H, Tashima Y, Zeniya A, Fujimori S, Otani S, Masamune O: Regulation of 60-kDa heat shock protein expression by systemic stress and 5-hydroxytryptamine in rat colonic mucosa. J Gastroenterol 29:721–726, 1994

    Google Scholar 

  40. Kuwabara T, Otaka M, Itoh H, Tashima Y, Zeniya A, Fujimori S, Otani S, Masamune O: Stress induces a 60-kDa heat shock protein expression in rat colonic mucosa. Gastroenterol Jpn 28:742, 1993

    Google Scholar 

  41. Koishi M, Hosokawa N, Sato M, Nakai A, Hirayoshi K, Hiraoka M, Abe M, Nagata K: Quercetin, an inhibitor of heat shock protein synthesis, inhibits the acquisition of thermotolerance in a human colon carcinoma cell line. Jpn J Cancer Res 83:1216–1222, 1992

    Google Scholar 

  42. Nakamura K, Rokutan K, Marui N, Aoike A, Kawai K: Induction of heat shock proteins and their implication in protection against ethanol-induced damage in cultured guinea pig gastric mucosal cells. Gastroenterology 101:161–166, 1991

    Google Scholar 

  43. Otaka M, Itoh H, Kuwabara T, Zeniya A, Fujimori S, Tashima Y, Masamune O: Functional difference between a 60 and 72 kDa heat shock proteins in the pancreas. Comparison of cytoprotective function in experimentally induced pancreatitis. Gastroenterology 108:A381, 1995

    Google Scholar 

  44. Emami A, Schwartz JH, Borkan SC: Transient ischemia or heat stress induced a cytoprotectant protein in rat kidney. Am J Physiol 260:479–485, 1991

    Google Scholar 

  45. Li GC, Li L, Liu RY, Rehman M, Lee WMF: Heat shock protein hsp70 protects cells from thermal stress even after deletion of its ATP-binding domain. Proc Natl Acad Sci USA 89:2036–2040, 1992

    Google Scholar 

  46. Kaplan MH: Acute and chronic pancreatitis. The clinical implications of their acceptance as disease of adaptation. Am J Gastroenterol 25:234–251, 1956

    Google Scholar 

  47. Nakai Y, Araki T, Takahashi S, Shimada A, Nakagawa T: Chronic pancreatitis as psychosomatic disorder. Psychother Psychosom 39:201–212, 1983

    Google Scholar 

  48. Takagi K, Kasuya Y, Watanabe K: Studies on the drugs for peptic ulcer. A reliable method for producing stress ulcer in rats. Chem Pharm Bull 12:465–472, 1964

    Google Scholar 

  49. Guth PH: Gastric blood flow in restraint stress. Am J Dig Dis 17:807–813, 1972

    Google Scholar 

  50. Fujimori S, Otaka M, Otani S, Jin M, Okuyama A, Itoh S, Iwabuchi A, Sasahara H, Itoh H, Tashima Y, Komatsu M, Masamune O: Induction of a 72-kDa heat shock protein and cytoprotection against thioacetamide-induced liver injury in rats. Dig Dis Sci 42:1987–1994, 1997

    Google Scholar 

  51. Viitanen PV, Lorimer GH, Seetharam R, Gupta RS, Oppenheim J, Thomas JO, Cowan NJ: Mammalian mitochondrial chaperonin 60 functions as a single toroidal ring. J Biol Chem 267:695–698, 1992

    Google Scholar 

  52. Backmann RP, Mizzen LA, Welch WJ: Interaction of hsp70 with newly synthesized proteins: Implications for protein folding and assembly. Science 248:850–854, 1990

    Google Scholar 

  53. Wiech H, Buchner J, Zimmermann R, Jakob U: Hsp90 Chaperons protein folding in vitro. Nature 358:169–170, 1992

    Google Scholar 

  54. Fujimori S, Otaka M, Itoh H, Kuwabara T, Zeniya A, Otani S, Tashima Y, Masamune O: Induction of a 60-kDa heat shock protein in rat gastric mucosa by restraint and water-immersion stress. J Gastroenterol 29:544–546, 1994

    Google Scholar 

  55. Welch WJ, Mizzen LA: Characterization of the thermotolerant cell. II. Effects on the intracellular distribution of heat-shock protein 70, intermediate filaments, and small nuclear ribonucleoprotein complex. J Cell Biol 106:1117–1130, 1988

    Google Scholar 

  56. Ohtani H, Wakui H, Komatsuda A, Satoh K, Miura AB, Itoh H, Tashima Y: Induction and intracellular localization of 90-kilodalton heat-shock protein in rat kidneys with acute gentamicin nephropathy. Lab Invest 72:161–165, 1995

    Google Scholar 

  57. Okuyama A, Otaka M, Itoh H, Jin M, Otani S, Itoh S, Iwabuchi A, Sasahara H, Tashima Y, Masanune O: Pre-induction of heat shock protein prevents development of L-arginine-induced necrotizing pancreatitis in rats. Gastroenterology 112:A470, 1997

    Google Scholar 

  58. Gething MJ, Sambrook J: Protein folding in the cell. Nature 355:33–45, 1992

    Google Scholar 

  59. Rothman JE: Polypeptide chain binding proteins: Catalysts of protein folding and related processes in cells. Cell 59:591–601, 1989

    Google Scholar 

  60. Hightower LE: Heat shock, stress proteins, chaperons, and proteotoxicity. Cell 66:191–197, 1991

    Google Scholar 

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Authors and Affiliations

  1. Department of Internal Medicine-1 and Department of Biochemistry, Akita University School of Medicine, Akita, Japan

    Akira Iwabuchi, Michiro Otaka, Setsuya Otani, Mario Jin, Atsushi Okuyama, Satoshi Itoh, Hideaki Sasahara, Masaru Odashima, Pacheco Ivan, Hideaki Itoh, Yohtalou Tashima, Osamu Masamune & Sumio Watanabe

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  1. Akira Iwabuchi
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  2. Michiro Otaka
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Iwabuchi, A., Otaka, M., Otani, S. et al. Specific Preinduction of 60-kDa Heat Shock Protein (Chaperonin Homolog) by TRH Does Not Protect Colonic Mucosa Against Acetic Acid-Induced Lesion in Rats. Dig Dis Sci 45, 1480–1489 (2000). https://doi.org/10.1023/A:1005597113024

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