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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15d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0h,0.5h,1h,1.5h,2h,2.5h,3h不同时间点取150μL细胞培养液与等量的100mg/LGriess液混合,室温下反应10min,在Σ960(λ=570nm)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,2h分泌达最高峰,3h渐进下降。其中0.5h与0h比较,差异有显著性(P<0.01);2h与0h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,2h达最高峰,3h渐进下降,而实验组A值基本保持不变。其中在2h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。
BACKGROUND: Numerous studies have confirmed that ginsenosides have a significant protective effect on the central nervous system, but it has not been reported to have the same protective effect on spinal nerves. Objective: To investigate the relationship between the effect of ginsenosides protecting spinal cord neurons and nitric oxide level and its mechanism. Design: Randomized controlled animal experiments. Unit: High-Pressure Oxygen Center of the PLA Navy General Hospital. Materials: The experiment was completed in 2000 at the Institute of Clinical Anatomy (National Key Laboratory) of the First Military Medical University of the Chinese People’s Liberation Army. There were 40 SD fetuses during the first 15 days of pregnancy. METHODS: Experiment 1: Spinal cord neurons of SD rat embryo were isolated and cultured. The spinal cord cell model was established with DMEM/F12 medium. On the fourth day after inoculation, the spinal cord axons were injured by the scratch method (simulated Nerve injury): Non-injured group was not treated, and 150 μL of cell culture medium was mixed with equal amount of 100 mg/LGries at 0h, 0.5h, 1h, 1.5h, 2h, 2.5h, and 3h after injury. The reaction was allowed to proceed for 10 min. The absorbance A value was measured on a microplate reader Σ960 (λ=570 nm). Experiment 2: Spinal cord neurons of SD rat embryo were also isolated and extracted. The experimental group was treated with ginsenoside + DMEM/F12 medium, and the control group was treated with DMEM/F12 medium. The absorbance was measured by the same method. MAIN OUTCOME MEASURES: 1 Relationship between spinal cord neuronal injury and nitric oxide levels. 2 The relationship between the protective effect of ginsenoside and nitric oxide level. RESULTS: 1 The relationship between spinal cord neuronal injury and nitric oxide levels: In the injured group, the secretion of nitric oxide increased after spinal cord nerve cell injury, peaked at 2 hours and decreased gradually at 3 hours. There was a significant difference between 0.5h and 0h (P<0.01), and there was a significant difference between 2h and 0h (P<0.01). 2 The relationship between the protective effect of ginsenoside and nitric oxide level: In the control group, the value of A increased with time, peaked at 2h, decreased gradually at 3h, while the value of A in the experimental group remained basically unchanged. At 2h, the difference between the two groups was statistically significant (P<0.01). CONCLUSION: The release of nitric oxide increases after peripheral nerve injury. Ginsenosides may be one of the ways to protect the peripheral nerves by inhibiting the release of nitric oxide.