CAJ | 학술논문

目的：探讨大鼠全脑缺血/再灌注介导的大鼠海马CA1区神经型一氧化氮合酶（ nNOS）巯基去亚硝基化对神经元损伤的作用及机制。方法将SD大鼠随机分为假手术组（ S组）、脑缺血/再灌注组（ I/R组）、给药组（ GSNO组、7-NI组、MK801组、NS102组、GYKI组）及溶剂对照组（生理盐水组、DMSO组）。采用四动脉结扎法构建大鼠全脑缺血/再灌注模型。运用生物素转化法检测蛋白质的巯基-亚硝基化，聚丙烯酰胺凝胶电泳、免疫印迹方法、焦油紫染色方法对nNOS巯基去亚硝基化水平以及大鼠海马神经元细胞的损伤进行研究。结果与S组相比，I/R组nNOS巯基亚硝基化水平明显降低（P＜0．05）；与I/R组相比，GSNO组、7-NI组以及MK801组nNOS巯基亚硝基化水平显著增高（P＜0．05），但GYKI组、NS102组、DMSO组以及生理盐水组与I/R组相比，差异无统计学意义。说明外源性一氧化氮（NO）供体GSNO、nNOS抑制剂7-NI、N-甲基-D-天冬氨酸（NMDA）受体抑制剂MK801能够抑制nNOS的去亚硝基化，对神经元起保护作用。结论大鼠全脑缺血/再灌注所致的大鼠海马CA1区nNOS巯基去亚硝基化在神经细胞损伤中有作用，其去亚硝基化是通过NMDA受体起作用；外源性NO供体GSNO、nNOS抑制剂7-NI也能够抑制nNOS的去亚硝基化从而对神经元起保护作用。
목적：탐토대서전뇌결혈/재관주개도적대서해마CA1구신경형일양화담합매（ nNOS）구기거아초기화대신경원손상적작용급궤제。방법장SD대서수궤분위가수술조（ S조）、뇌결혈/재관주조（ I/R조）、급약조（ GSNO조、7-NI조、MK801조、NS102조、GYKI조）급용제대조조（생리염수조、DMSO조）。채용사동맥결찰법구건대서전뇌결혈/재관주모형。운용생물소전화법검측단백질적구기-아초기화，취병희선알응효전영、면역인적방법、초유자염색방법대nNOS구기거아초기화수평이급대서해마신경원세포적손상진행연구。결과여S조상비，I/R조nNOS구기아초기화수평명현강저（P＜0．05）；여I/R조상비，GSNO조、7-NI조이급MK801조nNOS구기아초기화수평현저증고（P＜0．05），단GYKI조、NS102조、DMSO조이급생리염수조여I/R조상비，차이무통계학의의。설명외원성일양화담（NO）공체GSNO、nNOS억제제7-NI、N-갑기-D-천동안산（NMDA）수체억제제MK801능구억제nNOS적거아초기화，대신경원기보호작용。결론대서전뇌결혈/재관주소치적대서해마CA1구nNOS구기거아초기화재신경세포손상중유작용，기거아초기화시통과NMDA수체기작용；외원성NO공체GSNO、nNOS억제제7-NI야능구억제nNOS적거아초기화종이대신경원기보호작용。
Objective To investigate the denitrosylation of nNOS induced by cerebral ischemia /reperfusion on the apoptosis in the hippocampus CA 1 region of rats .Methods Transient cerebral ischemia for 15 min was performed in SD rats.The rats were randomly divided into 4 groups:sham group (S group), ischemia/reperfusion group (I/R group), u-nilateral intracerebroventricular injections of GSNO , 7-NI, MK801, NS102 and GYKI after ischemia/reperfusion groups (GSNO group, 7-NI group, MK801 grouop, NS102 group, and GYKI group), unilateral intracerebroventricular injec-tions 10%DMSO or saline groups ( DMSO group and saline group ) .The rat model of cerebral ischemia/reperfusion was established by 4-vessel occlusion method .S-nitrosylation was detected by the biotin -switch assay.SDS-PAGE, im-munoblotting and Cresyl violet staining were used to study the levels of denitrosylation of nNOS and the injury of the rat hippocampal neurons .Results The denitrosylation of nNOS was significantly noticeable in the I /R group than in the S group (P<0.05).The S-nitrosylation of nNOS was lightly observed in the GSNO group , 7-NI group and MK801 group than in the I/R group (P<0.05), meanwhile the GYKI group, NS102 group, DMSO group, and saline group were almost the same with I/R group.Conclusion The denitrosylation of nNOS plays an important role in the cell apop-tosis induced by I/R in the hippocampal CA 1 region of rats .