脑缺血/再灌注通过神经性一氧化氮合酶诱导KA受体亚基GluR6巯基亚硝基化的研究  

Cerebral ischemia-reperfusion induce GluR6 S-nitrosylation through neuronal NO synthase

作　　者：王梅 胡晓彤 戚大石 王蕾 张芳 

机构地区：[1]徐州医科大学形态学实验教学中心江苏省脑病重点实验室,江苏徐州221004 [2]徐州医科大学形态学实验教学中心,江苏徐州221004

出　　处：《中国当代医药》2016年第27期149-152,共4页China Modern Medicine

基　　金：国家自然科学基金青年基金项目(81500977);江苏省高校自然科学研究面上项目(14KJB180022);江苏省高校自然科学研究面上项目(13KJD310003);江苏省省级重点实验室开放研究课题(1505);江苏省徐州市科技计划项目(KC14SH076)

摘　　要：目的探讨脑缺血/再灌注是否通过神经性一氧化氮合酶(n NOS)诱导KA受体亚基Glu R6巯基亚硝基化。方法将28只SD大鼠随机分成4组,每组7只:假手术组、脑缺血/再灌注6 h组、脑缺血/再灌注6 h合并给药组、脑缺血/再灌注6 h溶剂对照组。采用四动脉结扎法构建大鼠全脑缺血模型,给予SD大鼠腹腔注射n NOS的抑制剂7-硝基吲唑。主要应用"生物素转化法"、SDS-PAGE、免疫印迹等方法对Glu R6的蛋白表达及其巯基亚硝基化水平进行研究。结果脑缺血/再灌注组与假手术组相比Glu R6的巯基亚硝基化水平显著升高,差异有统计学意义(P<0.05);脑缺血/再灌注联合n NOS的抑制剂7-NI组与脑缺血/再灌注组相比,Glu R6的巯基亚硝基化水平显著降低,差异有统计学意义(P<0.05);各组Glu R6的蛋白表达水平比较,差异无统计学意义(P>0.05)。结论在脑缺血/再灌注早期n NOS诱导Glu R6的巯基亚硝基化,为缺血性脑损伤疾病的治疗提供了新的理论依据。Objective To discuss whether neuronal nitric oxide synthase induce KA receptor subunit Glu R6 S-nitrosylation during ischemia-reperfusion.Methods 28 SD rats were randomly divided into 4 groups,each group had 7 rats：sham operation group,cerebral ischemia reperfusion 6 h combination with neuronal nitric oxide synthase inhibitor 7-NI group,cerebral ischemia reperfusion group,cerebral ischemia/reperfusion 6 h solvent control group.Transient cerebral ischemia was induced by a four-vessel occluded（4-VO） method.SD rats were treated intraperitoneally with 7-Nitroindazole, an inhibitor of n NOS（7-NI,25 mg/kg）.Biotin switch assay,SDS-PAGE and western blot was performed to study Snitrosylation and expression levels of Glu R6.Results Glu R6 S-nitrosylation levels in cerebral ischemia-reperfusion group increased significantly versus than that of sham group（P〈0.05）.Glu R6 S-nitrosylation levels in cerebral ischemiareperfusion combination with neuronal nitric oxide synthase inhibitor 7-NI group descended significantly versus cerebral ischemia-reperfusion group,it was significant difference（P〈0.05）;however,Glu R6 protein expression level was no significant difference among groups.Conclusion n NOS induce Glu R6 S-nitrosylating during the early stages of ischemia-reperfusion,which can be a new theory basis for stroke therapy.

关 键 词：谷氨酸受体6 脑缺血再灌注 巯基亚硝基化 神经性一氧化氮合酶 

分 类 号：R361.3[医药卫生—病理学]