Protective Effect of Trophic Factor Supplementation on Cold Ischemia/Rewarming Injury to Kidney Cells

Trophic factor supplementation에 의한 cold ischemia/rewarming손상으로 부터의 신장 세포 보호

  • Kwon, Young-Sam (School of Veterinary Medicine, University of Wisconsin-Madison) ;
  • Jang, Kwang-Ho (College of Veterinary Medicine, Kyungpook National University)
  • 권영삼 (위스콘신-매디슨 대학교 수의과대학) ;
  • 장광호 (경북대학교 수의과대학)
  • Published : 2008.10.31

Abstract

The aim of this study was to investigate whether trophic factor supplementation (TFS) enhance the survival of kidney cell during cold ischemic storage and rewarming. The effect of TFS on the phosphorylation of p44/42 and p38 mitogen activated protein kinases (MAPK) signaling pathway was determined by Western blot. Apoptotic changes after cold ischemic storage and rewarming was determined by 4',6'-diamino-2-phenylindole (DAPI) staining. The cell viability was evaluated by live assay. TFS significantly decreased p44/42 and p38 MAPK activity induced by cold ischemic injury and rewarming (p < 0.05). The number of apoptotic cells was decreased after 5 minute rewarming in the presence of TFS. TFS significantly increased the cell viability after 5 minute rewarming (p < 0.05). Therefore, it was concluded that trophic factor supplementation protects kidney tubule cells from cold ischemic and rewarming injury via the inhibition of p44/42 and p38 MAPK activation and reducing apoptotic change.

본 연구는 trophic factor supplementation (TFS)이 cold ischemic storage와 rewarming 동안에 신장 세포의 생존에 미치는 효과를 알아보기 위해 실시했다. p44/42와 p38 mitogen activated protein kinases (MAPK) 활성이 TFS에 의해 영향을 받는지를 Western blot을 통해 알아보았다. Apoptotic changes를 알아보기 위해 4',6'-diamino-2-phenylindole (DAPI) 염색을 실시했다. 세포생존도를 알아보기 위해 live assay를 실시하였다. 그 결과, TFS는 cold ischemic storage와 rewarming 동안 증가된 44/42와 p38 MAPK activity를 유의성 있게 감소시켰다 (p<0.05). 또한, cold ischemic storage와 rewarming에 의한 apoptotic cell 수가 TFS에 의해 감소함을 관찰했다. 마지막으로 TFS는 유의성 있게 세포 생존도를 증가시켰다 (p<0.05). 따라서, TFS는 p44/42와 p38 MAPK 활성을 감소시키고 apoptotic change를 억제함으로써 cold ischemia와 rewarming injury로부터 신장 세포를 보호하는 것으로 생각된다.

Keywords

References

  1. Hyman KM, Seghezzi G, Pintucci G, Stellari G, Kim JH, Grossi EA, Galloway AC, Mignatti P. Transforming growth factor-beta1 induces apoptosis in vascular endothelial cells by activation of mitogen-activated protein kinase. Surgery. 2002; 132: 173-179 https://doi.org/10.1067/msy.2002.125304
  2. Kruman II, Gukovskaya AS, Petrunyaka VV, Beletsky IP, Trepakova ES.Apoptosis of murine BW 5147 thymoma cells induced by cold shock. J Cell Physiol. 1992; 153: 112-117 https://doi.org/10.1002/jcp.1041530115
  3. Kwon YS, Foley JD, Murphy CJ, McAnulty JF. The effect of trophic factor supplementation on cold ischemia-induced early apoptotic changes. Transplantation. 2007; 83: 91-94 https://doi.org/10.1097/01.tp.0000242524.35562.4b
  4. Kwon YS, Foley JD, Russell P, McAnulty JF, Murphy CJ. Prevention of cold ischemia/rewarming-induced ERK 1/2, p38 kinase and HO-1 activation by trophic factor supplementation of UW solution. Cryobiology. 2008; 57: 72-74 https://doi.org/10.1016/j.cryobiol.2008.04.003
  5. McAnulty JF, Reid TW, Waller KR, Murphy CJ. Successful six-day kidney preservation using trophic factor supplemented media and simple cold storage. Am J Transplant. 2002; 2: 712-718 https://doi.org/10.1034/j.1600-6143.2002.20805.x
  6. Park KM, Chen A, Bonventre JV. Prevention of kidney ischemia/reperfusion-induced functional injury and JNK, p38, and MAPK kinase activation by remote ischemic pretreatment. J Biol Chem. 2001; 276: 11870-11876 https://doi.org/10.1074/jbc.M007518200
  7. Rauen U, Polzar B, Stephan H, Mannherz HG, de Groot H. Cold-induced apoptosis in cultured hepatocytes and liver endothelial cells: mediation by reactive oxygen species. FASEB J. 1999; 13: 155-168 https://doi.org/10.1096/fasebj.13.1.155
  8. Roberts JR, Rowe PA, Demaine AG. Activation of NFkappaB and MAP kinase cascades by hypothermic stress in endothelial cells. Cryobiology. 2002; 44: 161-169 https://doi.org/10.1016/S0011-2240(02)00018-4
  9. Roux PP, Blenis J, ERK and p38 MAPK-activated protein kinases: a family of protein kinases with diverse biological functions. Microbiol Mol Biol Rev. 2004; 68: 320-344 https://doi.org/10.1128/MMBR.68.2.320-344.2004
  10. Savill J, Mooney A, Hughes J. Apoptosis and renal scarring. Kidney Int Suppl. 1996; 54: 14-17
  11. Shankland SJ, Wolf G. Cell cycle regulatory proteins in renal disease: role in hypertrophy, proliferation, and apoptosis. Am J Physiol Renal Physiol. 2000; 78: 515-529
  12. Tanaka T, Miyata T, Inagi R, Kurokawa K, Adler S, Fujita T, Nangaku M. Hypoxia-induced apoptosis in cultured glomerular endothelial cells: involvement of mitochondrial pathways. Kidney Int. 2003; 64: 2020-2032 https://doi.org/10.1046/j.1523-1755.2003.00301.x
  13. Yilmaz A, Kliche S, Mayr-Beyrle U, Fellbrich G, Waltenberger J. p38 MAPK inhibition is critically involved in VEGFR-2-mediated endothelial cell survival. Biochem Biophys Res Commun. 2003; 306: 730-736 https://doi.org/10.1016/S0006-291X(03)01064-7