토끼 망막에서 NMDA에 의한 아포프토시스의 발현

• Published in: Daihan angwa haghoi jabji Vol. 49; no. 7; pp. 1146 - 1153
• Main Authors: 박용욱, Yong Wook Park, 채선화, Sun Hwa Chae, 이지웅, Ji Woong Lee, 권오주, Oh Ju Kwon, 신재필, Jae Pil Shin, 전창진, Chang Jin Jeon, 김시열, Si Yeol Kim
• Format: Journal Article
• Language: Korean
• Published: 대한안과학회 15.07.2008
• Subjects: Apoptosis; Bax; Bcl-2; Caspase-3; NMDA; 안과학; Bcl-2; Bax; NMDA; Caspase-3; Apoptosis
• ISSN: 0378-6471; 2092-9374

목적: 토끼 망막에서 NMDA에 의한 아포프토시스와 Bcl-2, Bax, caspase-3의 발현을 알아보고자 하였다. 대상과 방법: 20마리 토끼를 대상으로 680과 2,000 nmole의 NMDA를 유리체강내로 주입 후 2, 16, 60시간 및 1, 2주에 각각 안구를 적출하여 TUNEL 염색과 면역조직화학염색을 시행하였다. 결과: NMDA 주입 후 16시간 후에 신경절세포층 및 속핵층에서 TUNEL 염색에 양성을 보이는 세포가 가장 많이 나타났고 60시간에서 줄어들었고 2,000 nmole의 NMDA를 주입 후 680 nmole에 비해 TUNEL 양성세포 수가 더 많았다. Bcl-2, Bax, caspase-3의 면역조직화학염색에서도 신경절세포층과 속핵층의 세포가 염색이 되었고, 2시간에 이미 염색된 세포가 보였다. NMDA 주입 후 2시간에서 뮐러세포도 Bcl-2, Bax, caspase-3에 염색이 되었다. 결론: NMDA에 의한 망막신경세포의 흥분성 신경독성에 아포프토시스가 관여되며, 비록 대조군 실험은 하지 않았지만 Bcl-2, Bax, caspase-3의 발현이 apoptosis의 유발에 중요한 역할을 하며 뮐러세포도 관여할 것으로 생각된다. <한안지 49(7):1146-1153, 2008> Purpose: To evaluate the involvement of apoptosis in N-methyl-D-aspartate (NMDA)-induced excitotoxicity in the rabbit retina. Methods: After intravitreal injection of 680 and 2,000 nmoles of NMDA in rabbit eyes, the eyes were enucleated at 2, 16, and 60 hours and 1 and 2 weeks. The apoptotic cell death was determined with TdT-mediated biotin-dUTP nick end labeling (TUNEL) stain, and immunohistochemical stains of Bcl-2, Bax, and caspase-3 were performed. Results: TUNEL showed increased labeling scattered in the ganglion cell layer and inner nuclear layer from 16 to 60 hours. The number of TUNEL-positive nuclei decreased at 60 hours, and none was observed at 2 hours, 1 week, and 2 weeks. More TUNEL-positive nuclei were seen with injection of 2,000 nmoles compared to 680 nmoles. Bcl-2, Bax, and caspase-3 were seen histologically as early as 2 hours in the ganglion cell layer and inner nuclear layer; there was no stained nuclei with the TUNEL stain. At 2 hours after intravitreal NMDA injection, Bcl-2, Bax, and caspase-3 were also stained in Muller cells. Conclusions: This study showed that apoptosis is involved in NMDA-induced excitotoxicity in the rabbit retina. Bcl-2, Bax, and caspase-3 may play important roles in modulating the apoptosis in NMDA-induced excitotoxicity, and Muller cells are involved in the apoptotic pathway. J Korean Ophthalmol Soc 49(7):1146-1153, 2008