망막신경절세포에서 고장성 및 고오스몰 스트레스에 의한 긴장성반응유도 결합단백질의 발현
Expression of TonEBP by Hypertonic and Hyperosmolar Stress in RGC-5 Cells
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목적: 고장성(hypertonicity) 및 고오스몰 농도(hyperosmolarity) 환경에서 망막신경절세포(RGC-5)의 전사인자인 TonEBP (Tonicityresponsive enhancer binding protein)의 발현을 확인함으로써 망막신경절세포의 사멸을 방지하는 데 있어 TonEBP의 발현이 관여함을 알아보고자 하였다. 대상과 방법: RGC-5 세포 배양 후 고장성 스트레스를 유발하기 위해 50 mM NaCl, 100 mM mannitol, 50 mM 및 100 mM glucose를3, 6, 12, 24시간 처리한 후 Western immunoblotting 및 Real-time PCR을 시행하였다. 결과: 50 mM NaCl 처리 3, 6시간 및 100 mM mannitol 처리 6시간 후, 그리고 100 mM glucose 처리 3, 6시간 후에서 Westernblotting을 통해 TonEBP가 발현되는 것을 확인할 수 있었으며 real-time PCR을 통한 TonEBP의 발현은 50 mM NaCl 처리 3시간및 100 mM mannitol 처리 3, 24시간 후, 그리고 50 mM glucose 처리 3, 24시간 후에서 통계적 유의성을 보였다. 결론: Staurosporine으로 분화시킨 RGC-5 세포를 고장성 스트레스 환경에 노출시켰을 때 TonEBP가 발현되는 것을 단백질 및 mRNA수준에서 확인할 수 있었다.
Abstract
Purpose: In order to determine whether the Tonicity responsive enhancer binding protein (TonEBP) is expressed by hypertonicand hyperosmolar stress, TonEBP expression was investigated in the retinal ganglion cell (RGC) line, RGC-5 cells. Methods: After RGC-5 cells were cultured by Staurosporine, TonEBP expression was measured with Western immunoblottinganalysis and real-time reverse transcription-polymerase chain reaction in 50 mM NaCl, 100 mM mannitol, 50 mM glucose, or 100mM glucose at 3, 6, 12, and 24 hours after exposure to each environment. Results: In this study, the protein expression of TonEBP was determined to be statistically significantly checked in 50 mM NaClafter 3, and 6 hours, in 100 mM mannitol after 6 hours, and in 100 mM glucose after 3, and 6 hours. TonEBP messengerRibonucleic acid (mRNA) expression was determined to be statistically significantly checked in 50 mM NaCl after 3 hours, in 100mM mannitol after 3, and 24 hours, and in 50 mM glucose after 3, and 24 hours. Conclusions: These results suggested that TonEBP was expressed by hypertonic and hyperosmolar stress at the protein andmRNA levels. Further studies are nedded to determine the role of TonEBP and the mechanism of expression and regulation ofTonEBP.
참고문헌 (28)
- "The Advanced glaucoma intervention study (AGIS): 7. The relationship between control of intraocular pressure and visual field deterioration. The AGIS Investigators", Am J Ophthalmol, 130, pp.429~440 (2000)
- "Comparison of glaucomatous progression between untreated patients with normal-tension glaucoma and patients with therapeutically reduced intraocular pressures. Collaborative Normal-Tension Glaucoma study group", Am J Ophthalmol, 126, pp.487~497 (1998)
- Chen JZ, "A new clue to glaucoma pathogenesis", Am J Med, 114, pp.697~698 (2003)
- Flammer J, "Vascular dysregulation:a principal risk factor for glaucoma damage?", J Glaucoma, 8, pp.212~219 (1999)
- Matsumoto S, "Protein kinase C-gamma and calcium", calmodulin-dependent protein kinase II-alpha are persistently translocated to cell membranes of the rat brain during and after middle cerebral artery occlusion, 2004, pp.24, 54~61
- Speechly-Dick ME, "Protein kinase C. Its role in ischemic preconditioning in the rat", Circ Res, 75, pp.586~590 (1994)
- Piccoletti R, "State and activity of protein kinase C in postischemic reperfused liver", Exp Mol Pathol, 56, pp.219~228 (1992)
- Padanilam BJ, "Induction and subcellular localization of protein kinase C isozymes following renal ischemia", Kidney Int, 59, pp.1789~1797 (2001)
- Tanaka C, "The protein kinase C family for neuronal signaling", Annu Rev Neurosci, 17, pp.551~567 (1994)
- Chen L, "Opposing cardioprotective actions and parallel hypertrophic effects of delta PKC and epsilon PKC", Proc Natl Acad Sci USA, 98, pp.11114~11119 (2001)
- Whitlock NA, "Hsp27 upregulation by HIF-1 signaling offers protection against retinal ischemia in rats", Invest Ophthalmol Vis Sci, 46, pp.1092~1098 (2005)
- McNaughts KS, "Isoquinoline derivatives as endogenous neurotoxins in the etiology of Parkinson's disease", Biochem Pharmacol, 56, pp.921~933 (1998)
- Cunha-Vaz J, "Early breakdown of the blood-retinal barrier in diabetes", Br J Ophthalmol, 59, pp.649~656 (1975)
- King GL, "The cellular and molecular mechanisms of diabetic complications", Endocrinol Metab Clin North Am, 25, pp.255~270 (1996)
- Barber AJ, "Insulin rescues retinal neurons from apoptosis by a phophatidylinositol 3-kinase", Aktmediated mechanism that reduces the activation of caspase-3, 2001, pp.276, 32814~32821
- Hoffmann EK, "Membrane mechanisms and intracellular signaling in cell volume regulation", Int Rev Cytol, 161, pp.173~262 (1995)
- Nakayama Y, "The TonE", TonEBP pathway mediates tonicity-responsive regulation of UT-A urea transporter expression, 2000, pp.275, 38275~38280
- Woo SK, "TonEBP", NFAT5 stimulates transcription of HSP70 in response to hypertonicity, 2002, pp.22, 5753~5760
- Cha JH, "Hydration status affects nuclear distribution of transcription factor tonicity responsive enhancer binding protein in rat kidney", J Am Soc Nephrol, 12, pp.2221~2230 (2001)
- Lopez-Rodriguez C, "Bridging the NFAT and NF-kappaB families: NFAT5 dimerization regulates cytokine gene transcription in response to osmotic stress", Immunity, 15, pp.47~58 (2001)
- Zhang Z, "Expression of osmotic stress-related genesin tissues of normal and hypoosmotic rats", Am J Physiol Renal Physiol, 285, pp.F688~F693 (2003)
- Favale NO, "Hypertonic induction of COX2 expression requires TonEBP", NFAT5 in renal epithelial cells, 2009, pp.381, 301~305
- Roth I, "Osmoprotective transcription factor NFAT5", TonEBP modulates nuclear factor-kappaB activity, 2010, pp.21, 3459~3474
- Woo SK, "TonEBP", NFAT5 stimulates transcription of HSP70 in Response to Hypertonicity, 2002, pp.22, 5753~5760
- Lee SD, "TonEBP stimulates multiple cellular pathways for adaptation to hypertonic stress: organic osmolyte-dependent and -independent pathways", Am J Physiol Renal Physiol, 300, pp.F707~F715 (2011)
- Trama J, "The NFAT-related protein NFATL1 (TonEBP", NFAT5) is induced upon T cell activation in a calcineurin-dependent manner, 2000, pp.165, 4884~4894
- Jauliac S, "The role of NFAT transcription factors in integrin-mediated carcinoma invasion", Nat Cell Biol, 4, pp.540~544 (2002)
- Trama J, "The osmoprotective function of the NFAT5 transcription factor in T cell development and activation", J Immunol, 169, pp.5477~5488 (2002)