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背景:人参皂甙可以促智抗衰老,保护大脑皮质运动神经元,具有抗细胞凋亡作用,但作用机制不清。目的:观察人参皂甙Rb1和Rg1对许旺细胞神经生长因子表达的影响。设计、时间及地点:细胞学体外观察,于2004-03/06在深圳市第二人民医院完成。材料:成人新鲜离体神经取自创伤离断无法再植的肢体,由深圳市第二人民医院提供。人参皂甙Rb1、Rg1由长春白求恩医科大学提供。方法:剥去神经外膜,剪成1.0～2.0mm碎块,采用酶消化法分离许旺细胞,双30min差速黏附法去除成纤维细胞,获得纯净度达95%以上的许旺细胞。将纯化的许旺细胞按每孔10万个细胞接种在预先涂有多聚赖氨酸的96孔细胞培养板上,设立3组:人参皂甙Rb1组分别加入含量为10,20,40,60,80μg的人参皂甙Rb120μL;人参皂甙Rg1组分别加入含量为10,20,40,60,80μg的人参皂甙Rg120μL;对照组加入PBS液20μL。主要观察指标:流式细胞仪检测许旺细胞神经生长因子的表达率。结果:与对照组比较,培养48h后人参皂甙Rb1组、人参皂甙Rg1组许旺细胞神经生长因子表达率均明显升高(P<0.05),且呈剂量依赖性增加,当人参皂甙Rb1和Rg1质量浓度达60mg/L时许旺细胞神经生长因子表达率最高。人参皂甙Rb1及Rg1相同剂量组之间比较,许旺细胞神经生长因子表达率差异无显著性意义(P>0.05)。结论:人参皂甙Rb1和Rg1可以通过促进许旺细胞分泌神经生长因子而具有潜在加速周围神经损伤修复的作用。
BACKGROUND: Ginsenosides can promote anti-aging and protect cerebral cortex motor neurons and have anti-apoptosis effects, but the mechanism is unclear. Objective: To observe the effect of ginsenoside Rb1 and Rg1 on the expression of nerve growth factor in Schwann cells. DESIGN, TIME AND SETTING: The cytology in vitro observation was performed at Shenzhen Second People’s Hospital from March to June 2004. Materials: Adult fresh isolated nerves were obtained from limbs that could not be replanted after traumatization and were provided by Shenzhen Second People’s Hospital. Ginsenoside Rb1, Rg1 was provided by Changchun Bethune Medical University. Methods: The epineurium was stripped and cut into 1.0-2.0 mm pieces. Schwann cells were isolated by enzymatic digestion. Fibroblasts were removed by double 30 min differential adhesion method and Schwann cells with a purity of more than 95% were obtained. The purified Schwann cells were seeded with 100,000 cells per well in a 96-well cell culture plate pre-coated with polylysine, and 3 groups were set up: Ginsenoside Rb1 group was added in an amount of 10, 20, 40, 60 respectively. 80μg ginsenoside Rb 120μL; ginsenoside Rg1 group were added 10, 20, 40, 60, 80μg of ginsenoside Rg 120μL; control group was added PBS solution 20μL. MAIN OUTCOME MEASURES: Expression rate of nerve growth factor in Schwann cells was detected by flow cytometry. RESULTS: Compared with the control group, the expression rate of nerve growth factor in the ginsenoside Rb1 group and the ginsenoside Rg1 group was significantly increased (P<0.05) after 48 hours culture, and showed a dose-dependent increase, when the ginsenoside Rb1 and Rg1. At the concentration of 60mg/L, Schwann cells had the highest expression rate of nerve growth factor. Compared with the same dose group of ginsenoside Rb1 and Rg1, there was no significant difference in the expression rate of nerve growth factor in Schwann cells (P>0.05). CONCLUSION: Ginsenosides Rb1 and Rg1 have the potential to accelerate the repair of peripheral nerve injury by promoting Schwann cells to secrete nerve growth factor.