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目的探讨大鼠局灶性脑缺血2h再灌注6h神经元型一氧化氮合酶(neuronal nitric oxide synthase,nNOS)来源的一氧化氮(nitric oxide,NO)和过氧亚硝基阴离子(peroxymtrite,ONOO~-)对Caspase-3蛋白表达的影响。方法 48只健康雄性SD大鼠随机分为假手术组、单纯缺血/再灌注组、溶剂对照组和7-硝基吲唑(12.5、25、50mg/kg)组。闭塞大鼠左侧大脑中动脉造成局灶性脑缺血模型。给予选择性nNOS抑制剂7-硝基吲唑(12.5,25,50mg/kg)组,采用硝酸还原酶法检测脑组织NO含量、流式细胞术检测硝基酪氨酸表达、免疫组织化学法检测Caspase-3蛋白表达的变化。结果与假手术组相比,单纯缺血/再灌注组及溶剂对照组,脑组织NO含量,硝基酪氨酸含量及Caspase-3表达均升高(P<0.01)。与溶剂对照组比较,腹腔注入7硝基吲唑抑制nNOS活性可使Caspase-3蛋白表达降低(P<0.05)。结论大鼠局灶脑缺血再灌注过程中,nNOS来源的NO/ONOO可促进Caspase-3蛋白表达,这可能是nNOS活性增加促进细胞凋亡的机制之一。
Objective To investigate the effects of nitric oxide (NO) and peroxymtrite (NO) on the expression of neuronal nitric oxide synthase (nNOS) in rats after focal cerebral ischemia 2h, , ONOO ~ -) on Caspase-3 protein expression. Methods Forty-eight healthy male Sprague-Dawley rats were randomly divided into sham operation group, ischemia / reperfusion group, solvent control group and 7-nitroindazole group (12.5,25,50mg / kg). Occlusion of the left middle cerebral artery in rats resulted in a focal cerebral ischemia model. Nitric acid reductase was used to detect the content of NO in brain tissue, the expression of nitrotyrosine was detected by flow cytometry, the expression of nitrotyrosine was detected by immunohistochemistry Detection of Caspase-3 protein expression changes. Results Compared with the sham-operated group, NO content, nitrotyrosine content and Caspase-3 expression in the brain tissue were increased in the ischemia / reperfusion group and the solvent control group (P <0.01). Compared with the solvent control group, intraperitoneal injection of 7-nitro-indazole inhibited the activity of nNOS and decreased the expression of Caspase-3 protein (P <0.05). Conclusion NO / ONOO derived from nNOS can promote the expression of Caspase-3 during focal cerebral ischemia-reperfusion in rats. This may be one of the mechanisms of the increased nNOS activity in promoting apoptosis.