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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.
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.
