【摘要】 【目的】研究银杏叶标准提取物(EGb761)保护创伤脊髓运动神经元的细胞机制。【方法】成年Sprague-Dawley雄性大鼠行臂丛神经根撕脱术后,随机分为:撕脱组(生理盐水治疗)和EGb761组[EGb761100mg/(kg·d)治疗]。各组动物分别存活5d,2、4、6周后处死。硝酸还原酶法检测大鼠脊髓组织NO含量和cNOS酶活力,取C7节段切片行胆碱乙酰转化酶(ChAT)、神经元型一氧化氮合酶(nNOS)免疫组化、NADPH-d酶组化和中性红活细胞染色。【结果】EGb761组与撕脱组相比:不仅降低脊髓组织NO含量(μmol/g)(2周组0.24vs0.36、4周组0.22vs0.30、6周组0.19vs0.23;P均<0.05),也降低脊髓组织cNOS酶活力(U/mg)(5d组0.047vs0.057、2周组0.13vs0.20、4周组0.065vs0.14、6周组0.061vs0.083;P均<0.05)。EGb761恢复部分损伤运动神经元的ChAT活性、EGb761降低nNOS蛋白的异位表达,NADPH表达率(%)(2周组26.0vs40.7、4周组21.7vs36.9、6周组18.2vs26.7、8周组12.6vs20.9;P均<0.05);EGb761提高损伤运动神经元的生存率(%)(5d组92.2vs87.1、2周组77.1vs71.3、4周组56.8vs49.8、6周组48.0vs43.4、8周组31.3vs22.0;P均<0.05)。【结论】EGb761可能通过多靶点干扰损伤运动神经元的基因表达、蛋白酶活性以及代谢进程,减少臂丛撕脱伤诱导的运动神经元死亡。更多还原

【Abstract】 Objective To study the cellular mechanism of EGb761 effect on injured spinal motor neurons.Methods Male adult Sprague-Dawley rats were performed with roots avulsion of right brachial plexus under the microscope.Then the injured rats were randomly divided into avulsion groups(with normal saline treatment)and EGb761 groups with EGb761 100 mg/(kg·d)treatment.After survival for 5 days,2,4,6,or 8 weeks,the rats were killed,respectively.The NO and cNOS levels were detected by nitrate reductase method.The frozen sections of injured C7 spinal segments were prepared for ChAT,nNOS immunohistochemistry,NADPH-d enzyme stain and neutral red survival cell stain.Results Compared to avulsion groups,EGb761 not only decreased NO levels(μmol/g)(0.24 vs 0.36 at 2 week,0.22 vs 0.30 at 4 week,0.19 vs 0.23 at 6 week,with P < 0.05,respectively),but also decreased the activities of cNOS enzyme(U/mg)(0.047 vs 0.057 at 5 d,0.13 vs 0.20 at 2 week,0.065 vs 0.14 at 4 week,0.06 vs 0.083 at 6 week,with P < 0.05 respectively).In EGb761 treated rats,expression of ChAT were up-regulated and de novo nNOS expression were down-regulated with decreased amount of NADPH-positive motor neurons(%)(26.0 vs 40.9 at 2 week,21.7 vs 36.9 at 4 week,18.2 vs 26.7 at 6 week,12.6 vs 20.9 at 8 week,with P < 0.05 respectively);EGb761 enhanced the survival rate of injured motor neurons(%)(92.2 vs 87.1 at 5 d,77.1 vs 71.3 at 2 week,56.8 vs 49.8 at 4 week,48.0 vs 43.4 at 6 week,31.3 vs 22.0 at 8 week,with P < 0.05 respectively).Conclusion It is suggested that EGb761 may interfere with genomics,proteomics,and metabolic mechanism in root avulsion-induced spinal motor neuron neuropathy.更多还原