人参皂苷Rb1对体外培养雪旺细胞作用实验探究.docVIP

人参皂苷Rb1对体外培养雪旺细胞作用实验探究 　 作者：杨雷，刘黎军，肖建德，王大平，关瑞云 【摘要】 目的 研究人参皂苷Rb1对体外培养雪旺细胞增殖分化的影响，探讨其促进神经再生的作用和机制。方法 选用8个月月龄的新西兰兔的坐骨神经体外培养第2代，加入不同浓度的人参皂苷Rb1，继续培养10天，相差显微镜观察计数，绘制各自的增殖曲线；在36h和72h对各组细胞进行流式细胞分析。结果 活细胞计数含20μg/ml人参皂苷Rb1组的倍增时间为5.3天，明显优于对照组(P＜0.01)；用含20μg/ml人参皂苷Rb1培养36h、72h后雪旺细胞增殖的流式细胞仪分析，处于S期的雪旺细胞较对照组明显增高(P＜0.01)。结论 人参皂苷Rb1有促进体外培养雪旺细胞快速增殖分化的作用，从而为促进神经损伤的再生途径提供一些新的药物使用基础研究。 【关键词】 人参皂苷Rb1；雪旺细胞；细胞增殖 【Abstract】 Objective To study the effects of Ginsenoside Rb1 on the proliferation and differentiation of Schwanns cells and discuss the mechanism of nerve regeneration induced by Ginsenoside Rb1.Methods The sciatic nerve of New zealand rabbit of 8 months old was cultured in vitro and the second generation was used.Ginsenoside Rb1 with different concentrations were dropped into the culture medium for 10 days.Numbers of cells were counted by contrast microscope and proliferation curves were drawn individually.Furthermore，analysis of cells were made by flow cytometer.at 36h and 72h.Results Double proliferation time of Schwann’s cells cultured in 20μg/ml Ginsenoside Rb1 was 5.3d which superior to has was obvious difference from the control group(P＜0.01).The numbers of Schwanns cell at synthesis phase were increased significantly which cultured in 20μg/ml Ginsenoside Rb1(P＜0.01).Conclusion Ginsenoside Rb1 has the function of stimulating proliferation and idfferentiation of Schwann’s cell in vitro，and suqqests the important role in nerve regeneration. 【Key words】 Ginsenoside Rb1；schwanns cell；proliferation and differentiation of and suqqests 雪旺细胞是周围神经再生和周围神经组织工程研究的重要细胞，不仅为神经修复提供支撑，尚可诱导神经纤维生长的方向。近年来发现［1］尚有胶质细胞生长因子、成纤维细胞生长因子、霍乱毒素、血小板生长因子、转移生长因子以及cAMP类似物质等，均能刺激雪旺细胞生长，但是均有其局限性。应用中药治疗周围神经的损伤在临床上确有积极的疗效。为了解周围神经损伤后人参皂苷Rb1是否可以促使雪旺细胞增殖分化，从而促进周围神经损伤后的修复问题，我们设计了本实验研究。 1 材料与方法 1.1 药品与试剂 人参皂苷Rb1购自长春白求恩医科大学基础医学实验室。DMEM培养液、0.25%胰蛋白酶(含1mmolEDTA-4Na)、胶原蛋白酶和新生小牛血清为美国Gibco公司产品，L-多聚赖氨酸(相对分子质量为15万～30万，Sigma公司)；兔抗鼠S-100蛋白酶联免疫细胞化学试剂盒(北京中山生物公司)。 1.2 雪旺细胞的体外培养 选用8个月月龄的新西兰兔1只，于静脉麻醉下在双侧坐骨神经中段用1