유도된 열충격 단백질의 인간각막상피세포에 대한 보호효과

목적 : 안과에서 사용하는 적외선 온열요법은 상처치유를 빠르게 하는데, 이는 열충격 단백질이 유도되어 염증과 스트레스로 인해 세포 각막 상피 및 섬유화 세포에서 발생되는 세포고사를 억제하여 세포를 보호하기 때문으로 생각된다. 하지만 지금까지 이에 대한 과학적 실험근거가 없었던 바 저자들은 인간 각막상피세포에 glutamine 과 열충격을 주어 열충격 단백질의 발현 및 유도된 열충격 단백질의 세포 보호효과를 관찰하였다.<BR> 대상과 방법 : 인간 각막상피세포를 배양하여 대조군은 glutamine만을, 실험군은 glutamine과 열충격을 함께 준 후 열충격 단백질발현을 western blot과 면역염색으로 확인하였다. 유도된 열충격 단백질의 세포보호율은 etoposide로 세포를 고사시킨 후, MTT 방법으로 측정하였다.<BR> 결과 : Glutamine 단독 처리군 보다는 glutamine과 열충격을 같이 처리한 군에서 열충격 단백질의 발현이 많았고, 세포보호율도 높았다.<BR> 결론 : 열충격과 glutamine 처리 시 유도되는 열충격 단백질은 창상이나 염증 시 일어나는 세포 손상과 고사로부터 세포를 보호하는 것을 확인하였다.〈한안지 44(8):1879-1885, 2003〉

Purpose: The purposes of study were to assess the expression patterns of heat shock protein (HSP) after glutamine and glutamine with non- lethal heat shock treatment, to evaluate the protective effects of heat shock protein from apoptosis in cultured human corneal epithelial cell.<BR> Methods: The cultured human corneal epithelial cells were divided into two group. One group was treated with 0, 10, 20, 30, 40, 50 mM of glutamine and the other group was exposed to 43℃(heat shock) for 30 minutes with same concentration of glutamine. After glutamine and heat treatment, the expression patterns of Hsp 27, 70 were examined by western blot and immunohistochemistry. Apoptosis was induced with 80㎛ of etoposide. The viability (cell protection rate of heat shock protein) against apoptosis after etoposide treatment was measured by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay.<BR> Results: Expression of Heat shock protein 70 was not significantly effected in only glutamine treatment, but was remarkably increased in heat shock with glutamine treatment group. The increased cell number (viability=antiapoptotic effect of heat shock protein)of glutamine with heat shock group after etoposide treatment suggested that Hsp 70 appeared to be a major role in protection of Human corneal epithelial cell from apoptosis. The expression of Heat shock protein 27 was not effected in only glutamine and heat with glutamine treatment group.<BR> Conclusions: These data suggest that induced heat shock protein protect etoposide-generated apoptosis in human corneal epithelial cell. J Korean Ophthalmol Soc 44(8):1879-1885, 2003