Journal of Oral Medicine and Pain

Korean Academy of Orofacial Pain and Oral Medicine (대한안면통증구강내과학회)
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The Effects of Repeated Restraint Stress on the Rat Parotid Glands, Ultramicroscopical and Histochemical Study

구속 스트레스에 대한 백서 타액선 조직의 미세구조적, 조직화학적 변화

  • Yoon, In-Jong (Department of Orofacial Pain and Oral Medicine, School of Dentistry, Kyung Hee University) ;
  • Kang, Soo-Kyung (Department of Orofacial Pain and Oral Medicine, School of Dentistry, Kyung Hee University) ;
  • Auh, Q-Schick (Department of Orofacial Pain and Oral Medicine, School of Dentistry, Kyung Hee University) ;
  • Chun, Yang-Hyun (Department of Orofacial Pain and Oral Medicine, School of Dentistry, Kyung Hee University) ;
  • Hong, Jung-Pyo (Department of Orofacial Pain and Oral Medicine, School of Dentistry, Kyung Hee University)
  • 윤인종 (경희대학교 치의학전문대학원 안면통증구강내과학교실) ;
  • 강수경 (경희대학교 치의학전문대학원 안면통증구강내과학교실) ;
  • 어규식 (경희대학교 치의학전문대학원 안면통증구강내과학교실) ;
  • 전양현 (경희대학교 치의학전문대학원 안면통증구강내과학교실) ;
  • 홍정표 (경희대학교 치의학전문대학원 안면통증구강내과학교실)
  • Received : 2013.05.03
  • Accepted : 2013.05.28
  • Published : 2013.06.30
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Abstract

It has been known that saliva may affect the most of oral diseases. On the contrary, several systemic conditions may affect salivary flow and cause oral dryness and psychosocial stress especially may a crucial role in the etiology of hyposalivation and oral dryness. Many studies have focused on macroscopic effects of the stress on the salivary glands by autonomic response, but on the other hand it has hardly been reported on cellular microscopic effects of the stress on the salivary glands. Therefore, this study was performed to examine clusterin, a antiapoptotic and cytoprotective protein, in the parotid glands under restraint stress condition. For this study, 18 rats were divided into 3 groups; 1) 2 rats of group I were selected as a normal control. 2) 2 rats of group II, as a experimental control were placed in the restraint cone for 2 hours 3) 14 rats of group III were placed in the restraint cone for 2 hours once a day. The rats were sacrificed immediately(group II, as a experimental control), 1, 2, 3, 4, 5, 6 and 7 days after application of the stress and the both parotid glands were excised. Immunohistochemistry and electron microscopy were performed. The finding were as follows: 1. In parotid glands, repeated stress denaturalize the acinar cells, interacinous tissues and interacinous connective tissues were separated to individual acinar cells. After 4 days of experiment, there were lots of vacuoles and intercalated ducts. 2. In parotid glands, repeated stress make the rER which is in acinar cells swollen after 3 days of experiment and it was intensified to 4 days. After 5 days of experiment the edema got worse and degenerated. 3. In parotid glands, clusterin was reduced in ductal cell cytoplasm but in intercalated duct clusterin was slightly stained until 3 days prominently increased until 4 days and then decreased again after 5 days of experiment.

구강 내에 발생되는 질환의 대부분은 타액의 영향을 받는다. 타액분비에 영향을 주는 요소 중에서 스트레스에 광범위하게 노출되어 있는 현대인들에게 나타나는 타액분비장애는 매우 중요한 문제로 대두되고 있는데, 스트레스가 타액선에 미치는 영향에 관해서는 자율신경에 의한 반응에 대하여만 알려져 있고 세포수준의 미시적 변화에 대한 연구는 미흡한 실정이다. 이에 본 연구에서는 구속 스트레스 조건하에서 타액선 세포의 변화를 광학현미경과 전자현미경을 이용하여 구조적인 변화를 관찰하였고, 세포자사와 간접적으로 관련이 있다고 알려진 clusterin를 이용하여 백서 이하선의 변화를 관찰하고자 하였다. 실험재료로는 웅성백서 18마리를 사용하였으며, 정상대조군 2마리, 실험대조군 2마리, 실험군 14마리를 배정하였고, 실험 대조군과 실험군은 매일 2시간씩 restraint cone을 사용하여 구속스트레스를 부여하였다. 이들은 각 군별로 즉일(실험대조군), 1일, 2일, 3일, 4일, 5일, 6일, 7일에 각각 2마리씩 희생한 후 이하선 조직을 절취하여 전자현미경검사와 clusterin 단백질 검색을 위한 면역조직화학검사를 시행 한 바 다음과 같은 결과를 얻었다. 1. 광학현미경 하에서, 백서의 이하선은 구속스트레스에 의하여, 스트레스가 반복됨에 따라 선포세포, 선포엽간 조직, 선포간 결체조직이 변성되어, 선포 간 이개 현상이 나타났으며, 실험 4일 후에는 선포 내에 공포가 관찰되었고 많은 수의 선조관이 관찰되었다. 2. 전자현미경 하에서, 백서의 이하선은 구속스트레스에 의하여, 선포세포 내의 층판상 구조인 rER이 실험 3일 후에 부종이 시작되기 시작하였으며, 4일 후에 심화되었고, 5일 후에는 심하게 부종되어 변성된 상이 관찰되었다. 3. 백서의 이하선에서 clusterin은 구속스트레스에 의하여, 선조관의 세포질에서 소실되었고, 개재관에서는 3일 후까지는 미약하게, 4일 후에는 매우 증가되었다가, 실험 5일 후부터는 점차 감소되는 양상을 보였다.

Keywords

References

  1. 전양현, 홍정표. 스트레스와 구강질환. 대한신심스트레 스학회지. 1995;3(1):57-72.
  2. Kleinhauz IEM, Baht R and Littner M. Ante-cedents of burning mouth syndrome- recent life event vs. psychopathologic aspects. J Dent Res. 1994;73: 567-571.
  3. Martin SG, Michael G, Jonathan A. Burket's oral medicine, 11th ed. 2008. BC Decker.
  4. 전양현, 홍정표. 스트레스가 streptozotocin유도 당뇨백 서 타액선의 clusterin(SGP-2) 변화에 미치는 영향에 관한 분자생물학적 연구. 대한신심스트레스학회지. 1997;5(2):13-32.
  5. Fritz IB and Murphy B. Clusterin. Insights into a multifunctional protein. Trends Endocrinol Metub. 1993;4:41-45. https://doi.org/10.1016/S1043-2760(05)80013-X
  6. Jenne DE and Tschopp J. Molecular structure and functional characterization of a human complement cytolysis inhibitor found in blood and seminal plasma: Identity to sulfated glycoprotein 2, a constituent of rat testis fluid. Proc Natn Acad Sci USA. 1989;86: 7123-7127. https://doi.org/10.1073/pnas.86.18.7123
  7. Jenne DE and Tschopp J. Clusterin: the intriguingguises of a widely expressed glycoprotein. TIES. 1992;17:154-159.
  8. Mark ER, John S. Clusterin: Physiologic and pathophysiologic considerations. Int J Biochem Cell Biol. 1995;27(7):633-645. https://doi.org/10.1016/1357-2725(95)00027-M
  9. 김흥모, 전양현, 홍정표. 스트레스가 streptozotocin유도 당뇨백서의 타액선에 미치는 영향. 대한구강내과학회 지. 1997;22(1):65-80.
  10. 김영준. 스트레스와 정신의학. 대한신심스트레스학회지. 1993;1(1):97-102.
  11. Tanaka M, Kohno Y, Nakagawa R, Ida Y, Takeda S and Nagasaki N. Time-related differences in noradrenaling turnover in rat brain regions by stress. Pharmacol Biochem Behav. 1982;16:315-319. https://doi.org/10.1016/0091-3057(82)90166-6
  12. Nukina I, Glavin GB and Labella FS. Acute coldresistant stress affects alpha 2-adrenoreceptors in specific brain regions of the rat. Brain Res. 1987;401: 30-33. https://doi.org/10.1016/0006-8993(87)91159-0
  13. Finkelstein Y, Koffler B, Rabey JM and Gilad GM. Dynamics of cholinergic sympathetic mechanisms in rat hippocampus after stress. Brain Res. 1985;343: 314-319. https://doi.org/10.1016/0006-8993(85)90749-8
  14. Yoneda Y, Kanmori K, Ida S and Kuriyama Y. Stress-induced alteration in metabolism of r-aminobutyric acid in rat brain. J Neurochem. 1983;20:350.
  15. Dudar JD. The effect of septal nucleus stimulation on the release of acetylcholin from the rabbit hippocampus. Brain Res. 1975;83:123-133. https://doi.org/10.1016/0006-8993(75)90863-X
  16. Guyton. Textbook of medical physiology. 8th ed. Philadelphia. 1991. WB Saunders Co. pp.848.
  17. Arancibia S, Tapia-Arancibia L, Assenmacher I and Astier H. Direct evidence of short-term cold-induced TRH release in the median eminance of unanesthetized rats. Neuroendocrinology. 1983;37:225-228. https://doi.org/10.1159/000123547
  18. Shires GT. Important factors in the maintenance fo homeostasis in the surgical patient. Acta Chir Scand (Suppl). 1988;550:29-35.
  19. Cannon WB. The emergency function of the adrenal medulla in pain and the major emotions. Am J Physiology. 1914;33:356-372.
  20. Frankenhaeuser M. Psychoneuroendocrine approaches to the study of the emotion as related to stress and coping. In H.E. Howe & R. A. Dienstibier(eds.), Nebraska symposium on motivation 1978 Lincon. 1979. University of Nebraska Press.
  21. Frankenhaeuser M and Lundberg U. Sympatheticadrenal and pituitary-adrenal response to challenge. In P Pichot, P Berner, R Wolf & K Thau(Eds.), Psychiatry, Vol. 2, London. 1985. Plenum
  22. Chrousos GP. Stressor, stress and neuroendocrinology intergration of the adaptive response. The 1997 Hans Selye Memorial lecture. Ann NY Acad Sci. 1998;851:311-335 https://doi.org/10.1111/j.1749-6632.1998.tb09006.x
  23. Ader R, Cohen N. Psychoneuroimmunology: conditioning and stress. Annu Rev Psycho. 1993;44:53-85. https://doi.org/10.1146/annurev.ps.44.020193.000413
  24. McEwen BS, Stellar E. Stress and the individual: mechanisms leading to disease. Arch Intern Med. 1993;153:2093-2101. https://doi.org/10.1001/archinte.1993.00410180039004
  25. Chrousos GP. The hypothalamic-pituitary-adrenal axis and immune-mediated inflammation. N Engl J Med. 1995;332:1351-1362. https://doi.org/10.1056/NEJM199505183322008
  26. Calabrase JR, Kling MA and Gold PW. Alteration in immunocompetence during stress, bereavement and depression: Focus on neuroendocrine regulation. Am J Psychiatry. 1987;144(9):1123-1134. https://doi.org/10.1176/ajp.144.9.1123
  27. Kiecolt-Glaser JK, Garner W, Speicher C, Penn GM, Holliday J and Glaser R. Psychosocial modiffiers of immunocompetence in medical students. Psychosomatic Medicine. 1984;46(1):7-14. https://doi.org/10.1097/00006842-198401000-00003
  28. Macleod RI. Psychologic factors in oral lichen planus. Br Dent J. 1992;173(3):88.
  29. Redman RS, Vance FL, Gorlin RJ, Peagler FD and Meskin LH. Psychological component in the etiology of geographic tongue. J Dent Res. 1966;45:1403-1408. https://doi.org/10.1177/00220345660450052301
  30. Bierman SM. A retrospective study of 375 patients with genital herpes simplex infections seen between 1973 and 1980. Cutis. 1983;31:548-565.
  31. Browning S, Hislop S Scully C and Shirlaw P. The association between burning mouth syndrome and psychosocial disorders. Oral surg. 1987;64:171-174. https://doi.org/10.1016/0030-4220(87)90085-5
  32. Blaschuk O, Burdzy K and Fritz IB. Purification and characterization of a cell-aggregating factor (clusterin), the major glycoprotein in ram rete testis fluid. J Biol Chem. 1983;258:7714-7720.
  33. Jones SE, Jomary C. Molecules in focus clusterin. Int J Biochem Cell Biol. 2002;34: 427-431. https://doi.org/10.1016/S1357-2725(01)00155-8
  34. Sally AL, Philip EM. Clusterin expression during programmed and teratogen-induced cell death in the postimplantation rat embryo. Teratology. 1995;52:41- https://doi.org/10.1002/tera.1420520106
  35. Fritz IB, Murphy B. Insight into a multifunctional protein. TEM, 1993;4(2):41-45.
  36. Burkey BF, deSilva HV and Harmony JAK. Intracellular processiong of apolipoprotein J precursor to the mature heterodimer. J Lipid Res. 1991;32:1039- 1048.
  37. Jordan-Starck TC, Witte DP, Aronow BJ and Harmony JA. Apolipoprotein J: a membrane policeman. Curr Opin Lipidol. 1992;3:75-85. https://doi.org/10.1097/00041433-199204000-00004
  38. Murphy BF, Kirszbaum L, Walker ID and d''Apice AJF. SP-40,40 a newly identified normal human serum protein found in the SC5b-9 complex of complement and in the immune deposits of glomerulonephritis. J Clin Invest. 1988;81:1858-1864. https://doi.org/10.1172/JCI113531
  39. Kirszbaum L, Sharpe JA, Murphy B, d''Apice AJF, Classon B, Hudson P and Walker ID. Molecular cloning and characterization of the novel, human complement-associated protein, SP40, 40: a link between the complement and reproductive systems. EMBO J. 1989;8:711-718.
  40. Willson MR and Easterbrook-Smith SB. Clusterin binds by a multivalent mechanism to the Fc and Fab regions of IgG. Biochem Biophys Acta. 1992;1159: 319-326.
  41. Ehnholm C, Bozas SE, Tenkanen H, Kirszbaum L, Metso J, Murphy B and Walker ID. The apolipoprotein A-l binding protein of placenta and the SP-40,40 protein of human blood are different proteins which both bind to apolipoprotein A-l. Biochim Biophys Acta. 1991;1086:225-260. https://doi.org/10.1016/0005-2760(91)90011-6
  42. Sylvester SR, Morales C, Oko R and Griswold MD. Localization of sulfated glycoprotein-2 (clusterin) on spermatozoa and in the reproductive tract of the male rat. Biol Reprod. 1991;45:195-207. https://doi.org/10.1095/biolreprod45.1.195
  43. Purrello M, Bettuzzi S, Di Pietro C, Mirabile E, Di Blasi M, Rimini R, Grzeschik KH, Ingletti C, Corti A and Sichel G. The gene for SF40,40, human homolog of rat sulfated glycoprotein 2, rat clusterin, and rat testosterone-repressed prostate message 2, maps to chromosome 8. Genomics. 1991;10:151-156. https://doi.org/10.1016/0888-7543(91)90495-Z
  44. Leger JG, Montpetit ML and Tenniswood MP. Characterization and cloning of androgen-repressed mRNAs from rat ventral prostate. Biochem biophys Res Commun. 1987;147:196-203. https://doi.org/10.1016/S0006-291X(87)80106-7
  45. Jun HO, Kim DH, Lee SW, Lee HS, Seo JH, Kim JH, Kim JH, Yu YS, Min BH and Kim KW. Clusterin protects H9c2 cardiomyocytes from oxidative stress-induced apoptosis via Akt/GSK-3盃signaling pathway. Exp Mol Med. 2011;43(1):53-61. https://doi.org/10.3858/emm.2011.43.1.006
  46. Kim NY and Choi WS. Proapoptotic role of nuclear clusterin in brain. Anat Cell Biol. 2011;44:169-175. https://doi.org/10.5115/acb.2011.44.3.169
  47. Klock G, BaiersdÖrfer M and Koch-Brandt C. Chapter 7;Cell protective functions of secretory Clusterin(sCLU). Adv Cancer Res. 2009;104:115-138. https://doi.org/10.1016/S0065-230X(09)04007-X
  48. Falgarone G and Choicchia G. Chapter 8;Clusterin; A multifacet protein at the crossroad of inflammation and autoimmunity. Adv Cancer Res. 2009;104: 139-170. https://doi.org/10.1016/S0065-230X(09)04008-1