BACKGROUND: Researches suggest that ginseng saponin (GS) has protective effect on central nerve, but the effect on spinal nerves is reported rarely.OBJECTIVE: To investigate the relationship between effect of GS on spinal nerve and level of nitrogen monoxide (NO) and its mechanism.DESIGN: Randomized controlled animal study.SETTING: Military Hyperbaric Oxygen Center of Navy General Hospital of Chinese PLA.MATERIALS: The experiment was completed at Clinical Anatomy Institute (National Key Laboratory) of the First Military Medical University of Chinese PLA in 2000. Forty SD foetus rats with 15-day conception were selected.METHODS: Study Ⅰ: Embryo-spinal nerve cells of SD rats were separated, extracted and modeled with DMEM/F12 culture medium. On the fourth day of inoculated culture, axon of spinal nerve (simulation of peripheral nerve injury) was damaged with scarification method in injury group, but that in non-injury group was not treated. 150 μL cell culture medium and 100 mg/L Griess solution were mixed at 0, 0.5 1, 1.5, 2, 2.5, and 3 hours after injury respectively. Absorbency (A) was assayed with Σ960 (λ=570 nm) enzyme-linked immunoadsordent assay (ELISA) symbolic device 10 min-utes after reaction at room temperature. Study Ⅱ: Embryo-spinal nerve cells of SD rats were separated and extracted. Those in the experimental group were treated with GS + DMEM/F12 culture medium, but with DMEM/F12 culture medium in the control group. A value was assayed with the same method.MAIN OUTCOME MEASURES:① Relationship between injury of spinal neurons and level of NO;② Relationship between protective effect of GS and level of NO.RESULTS:① Relationship between injury of spinal neurons and level of NO: In the injury group, NO secretion was increased after injury of spinal neurons, reached peak 2 hours later, and decreased 3 hours later. There was significant difference as compared 0.5 hour with 0 hour (P ＜ 0.01),and also there was significant difference as compared 2 hourswith 0 hour (P ＜ 0.01).② Relationship between protective effect of GS and level of NO: In the control group, A value was increased with time passing, reached peak 2 hours later, and decreased 3 hours later; but A value in the experimental group was not changed generally. There was significant difference between the two groups at 2-hour point (P ＜ 0.01).CONCLUSION: NO liberation is increased after peripheral nerve injury.GS can inhibit NO liberation so as to protect peripheral nerve.%背景:众多的实验证实人参皂甙对中枢神经有明显的保护作用,但其对脊髓神经是否具有同样的保护作用,报道较少.目的:探讨人参皂甙保护脊髓神经元的作用与一氧化氮水平的关系及其机制.设计:随机对照动物实验.单位:解放军海军总医院全军高压氧中心.材料:实验于2000年在解放军第一军医大学临床解剖研究所(国家重点实验室)完成.孕期15 d的SD胎鼠40只.方法:实验1:分离提取SD鼠胚脊髓神经细胞,用DMEM/F12培养基,建立培养的脊髓细胞模型,在接种培养的第4天,损伤组采用划痕法损伤脊髓细胞轴突(模拟周围神经损伤),非损伤组不进行处理,分别在损伤后0 h,0.5 h,1 h,1.5 h,2 h,2.5 h,3 h不同时间点取150 μL细胞培养液与等量的100 mg/L Griess液混合,室温下反应10 min,在∑960(入=570 nm)酶标仪上测定吸光度A值.实验2:同样分离提取SD鼠胚脊髓神经细胞,实验组用人参皂甙+DMEM/F12培养基,对照组用DMEM/F12培养基,相同方法测定吸收度.主要观察指标:①脊髓神经元损伤与一氧化氮水平的关系.②人参皂甙保护作用与一氧化氮水平的关系.结果:①脊髓神经元损伤与一氧化氮水平的关系:在损伤组中,脊髓神经细胞损伤后一氧化氮分泌增加,2 h分泌达最高峰,3 h渐进下降.其中0.5 h与0 h比较,差异有显著性(P＜0.01);2 h与0 h比较,差异有显著性(P＜0.01).②人参皂甙保护作用与一氧化氮水平的关系:在对照组中,A值随着时间变化而变化增加,2 h达最高峰,3 h渐进下降,而实验组A值基本保持不变.其中在2 h时间点,两组比较,差异有显著性(P＜0.01).结论:周围神经损伤后释放一氧化氮增加,人参皂甙通过抑制一氧化氮释放,可能是其保护周围神经的途径之一.
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