Applied Microscopy

  • 제37권3호
  • /
  • Pages.167-174
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  • 2007
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  • 2287-5123(pISSN)
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  • 2287-4445(eISSN)
한국현미경학회 (Korean Society of Microscopy)
권호 표지

콩팥에서 Erythropoietin 투여로 인한 HSP70의 발현 변화

Expression of HSP70 Immunoreactivity in EPO Treated Rat Kidney

  • 정주영 (충남대학교 수의과대학 해부학교실) ;
  • 김진 (가톨릭대학교 의과대학 해부학교실)
  • Jung, Ju-Young (Department of Veterinary Anatomy, College of Veterinary Medicine, Chungnam National University) ;
  • Kim, Jin (Department of Anatomy, College of Medicine, The Catholic University of Korea)
  • 발행 : 2007.09.30
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초록

Heat shock protein 70 (HSP70)은 다양한 질병상태와 치명적인 열 손상에서 세포 및 조직을 보호하는데 중요한 역할을 하며, 또한 외부의 stress로부터 세포내 단백질의 파괴와 변화를 감소시키는 단백질로 알려져 있다. 본 연구에서는 오랜 기간 동안 조혈기관의 치료제로 콩팥에서도 세포보호효과가 있는 것으로 알려진 Erythropoietin(EPO)을 투여하여 콩팥내의 HSP70의 발현변화를 세포수준에서 관찰하고자 하였다. Sprague-Dawley계 흰쥐를 사용하여 전자현미경적 면역조직화학법으로 rHuEPO투여군과 대조군에서 HSP70의 발현변화를 관찰하였다. 대조군에서 HSP70은 콩팥의 바깥수질과 속수질에서 관찰되었으며, 특히 속수질에서 강하게 발현되었고 그 부위는 속수질집합관세포와 헨레고리의 내림가는 부분이었다. EPO 투여군에서는 속수질과 바깥수질의 내림가는 부분에서는 발현변화가 관찰되지 않았으나, 바깥수질의 집합관 세포에서 발현이 급격히 증가함을 관찰되었다. 특히 대조군에서의 핵주변부위뿐 아니라 세포내 핵상부분을 비롯한 세포막주변부위에도 강한 면역 염색성을 나타내었다. 이러한 결과는 콩팥의 바깥수질에서 stress성 단백질인 HSP70의 조절기전이 EPO에 의해 매개됨을 보여주면, 세포 stress및 질병상태에서도 이러한 기전이 작용할 것으로 생각된다.

Heat shock protein (HSP) 70 functions as a molecular chaperon and reduces stress-induced denaturation and aggregation of intracellular proteins. Erythropoietin (EPO) plays an important role during acute renal failure repair process by rapidly correcting anemia and enhancing renal tubular regeneration. The purpose of this study was to examine the effect of EPO treatment on renal HSP70 expression. Male Sprague-Dawley rats were injected rHUEPO. Kidney were preserved by in vivo perfusion with paraformaldehyde-lysine-periodate (PLP) and processed for immunohistochemistry and electron microscopy. In control kidney, HSP70 was expressed in the cortex, outer medulla and inner medulla. Especially, HSP immunoreactiviy was mainly founded in descending thin limb of outer medulla and inner medullary collecting duct. In EPO treated kidney, HSP70 expression markedly increased in the descending thin limb of outer medulla and newly detected in cortical collecting duct. Electron microscopy showed the presence of HSP immunoreactivity on the intracelluar vesicles and Golgi complex of descending thin limb and cortical collecting duct. These findings suggest that EPO treatment leads to new production of HSP70 in renal tubular cells, and induction of HSP70 by rHuEPO is causally related to protective function.

키워드

참고문헌

  1. Bernaudin M, Marti HH, Rouseel S, Divoux D, Nouvelot A, MacKenzie ET, Petit E: A potential role for erythropoietin in focal permanent cerebral ischemia in mice. J Cereb Blood Flow Metab 19 : 643-651, 1999 https://doi.org/10.1097/00004647-199906000-00007
  2. Brines ML, Ghezzi P, Keenan S, Angello D, de Lanerolle NC, Cerami C, Itri LM, Cerami A: Erythropoietin crosses the blood-brain barrier to protect against experimental brain injury. Proc Natl Acad Sci USA 97 : 10526-10531, 2000
  3. Buzzard KA, Giaccia AJ, Killender M, Anderson RL: Heat shock protein 72 modulates pathways of stress-induced apoptosis. J Biol Chem 273 : 17147-17153, 1998 https://doi.org/10.1074/jbc.273.27.17147
  4. Digicaylioglu M, Lipton SA: Erythropoietin-mediated neuroprotection involves cross-talk between Jak2 and NF-kappa- B signalling cascades. Nature 412 : 641-647, 2001 https://doi.org/10.1038/35088074
  5. Gabai VL, Meriin AB, Mosser DD, Caron AW, Rits S, Shifrin VI, Sherman MY: HSP70 prevents activation of stress kinases. A novel pathway of cellular thermotolerance. J Biol Chem 272 : 18033-18037, 1997 https://doi.org/10.1074/jbc.272.29.18033
  6. Hendrick JP, Hartl FU: Molecular chaperone functions of the heat-shock proteins. Annu. Rev. Biochem. 62 : 349-384, 1993 https://doi.org/10.1146/annurev.bi.62.070193.002025
  7. Jaattela M, Wissing D, Bauer PA, Li GC: Major heat shock protein hsp70 protects tumor cells from tumor necrosis factor cytotoxicity. EMBO J 11 : 3507-3512, 1992
  8. Kultz D: Hyperosmolality triggers oxidative damage in kidney cells. Proc Natl Acad Sci USA 101(25) : 9177-9178, 2004
  9. Madamanchi NR, Li S, Patterson C, Runge MS: Reactive oxygen species regulate heat-shock protein 70 via the JAK/STAT pathway. Arterioscler Thromb Vasc Biol 21 : 321-326, 2001 https://doi.org/10.1161/01.ATV.21.3.321
  10. Mestril R, Chi SH, Sayen MR. O' Reilly K, Dillmann WH: Expression of inducible stress protein 70 in rat heart myogenic cells confers protection against stimulated ischemiainduced injury. J Clin Invest 93: 759-767, 1994 https://doi.org/10.1172/JCI117030
  11. Miyake T, Kung CK, Goldwasser E: Purification of human erythropoietin. J Biol Chem 252 : 5558-5564, 1977
  12. Nemoto T, Yokota N, Keane WF, Rabb H: Recombinant erythropoietin rapidly treats anemia in ischemic acute renal failure. Kidney Int 59 : 246-251, 2001 https://doi.org/10.1046/j.1523-1755.2001.59780246.x
  13. Plumier JC, Ross BM, Currie RW, Angelidis CE, Kazlaris H, Kollias G, Pagoulatos GN: Transgenic mice expression the human heat shock protein70 have improved post-ischemic myocardial recovery. J Clin Invest 95 : 1854-1860, 1995 https://doi.org/10.1172/JCI117865
  14. Sadamoto Y, Igase K, Sakanaka M, Sato K, Otsuka H, Sakaki S, Masuda S, Sasaki R: Erythropoietin prevents place navigation disability and cortical infarction in rats with permanent occlusion of the middle cerebral artery. Biochem Biophys Res Commun 253 : 26-32, 1998 https://doi.org/10.1006/bbrc.1998.9748
  15. Vairano M, Dello Russo C, Pozzoli G, Battaglia A, Scambia G, Tringali G, Aloe-Spiriti MA, Preziosi P, Navarra P: Erythropoietin exerts anti-apoptotic effects on rat microglial cells in vitro. Eur J Neurosci 16 : 684-692, 2002 https://doi.org/10.1046/j.1460-9568.2002.02116.x
  16. Vaziri ND, Zhou XJ, Liao SY: Erythropoietin enhances recovery from cisplatin-induced acute renal failure. Am J Physiol 266 : 360-366, 1994 https://doi.org/10.1152/ajpcell.1994.266.2.C360
  17. Wang YH, Knowlton AA, Li FH, Borkan SC: Hsp72 expression enhances survival in adenosine triphosphate-depleted renal epithelial cells. Cell Stress Chaperones 7 : 137-145, 2002 https://doi.org/10.1379/1466-1268(2002)007<0137:HEESIA>2.0.CO;2