摘要目的 探讨人参皂甙Rb1对嗅鞘细胞(OECs)增殖及神经生长因子(NGF)、脑源性神经营养因子(BDNF)、胶质源性神经营养因子(GDNF)和神经细胞黏附分子(N-CAM)的表达和分泌等细胞生物学活性的影响,从而调节OECs对脊髓损伤的修复作用.方法 从成年SD大鼠嗅球取材获得OECs体外培养、纯化,四甲基偶氮噻唑(MTT)法测定人参皂甙Rb1在0,10,20,40,80 μg/ml浓度梯度下分别干预12,24,36,48,60 h对OECs增殖的影响,得出人参皂甙Rb1对OECs 作用的最适浓度和干预时间,并以此最适浓度和干预时间继续实验:细胞分成空白对照组和人参皂甙Rb1最适浓度和干预时间组(人参皂甙Rb1组),RT-PCR检测两组细胞NGF、BDNF、GDNF及N-CAM的mRNA表达,ELLSA法检测两组培养上清液中NGF、BDNF及GDNF分泌量.结果 MTT结果显示,人参皂甙Rb1不同浓度均能促进OECs增殖,与0μg/ml比较,20 μg/ml作用48 h增殖效应达峰值[(0.648 ±0.019)∶(0.547 ±0.024)] (P ＜0.05).RT-PCR结果显示,人参皂甙Rb1组较对照组显著上调OECs NGF、BDNF、GDNF及N-CAM的mRNA表达[(0.620±0.011)∶ (0.180 ±0.011)、(0.511 ±0.090)∶ (0.293 ±0.051)、(0.343 ±0.042)∶ (0.064 ±0.005)、(0.839 ±0.017)∶(0.717 ±0.044)] (P ＜0.05).ELISA结果显示,人参皂甙Rb1组较对照组显著促进OECs NGF、BDNF及GDNF的分泌[(200.167 ±8.361)∶(51.467 ±3.815)、(156.700 ±4.190)∶(96.500±2.707)、(26.264±5.864)∶(4.917±10.894)] (P ＜0.05).结论 人参皂甙Rb1对OECs增殖及相关因子的表达和分泌等生物学活性具有显著促进作用,从而有利于脊髓损伤的修复.

abstractsObjective To investigate influence of ginsenoside Rb1 on the proliferation and bioactivity of olfactory ensheathing cells (OECs).Methods OECs were primary cultured and purified from olfactory bulb of the adult SD rats.MTT assay was used to detect proliferation of OECs treated with ginsenoside Rb1 (intervention concentrations of 0,10,20,40,and 80 μg/ml and intervention time of 12,24,36,48,and 60 hours).Optimal concentration and intervention time of ginsenoside Rb1 was determined and performed in the succedent experiments.Purified cells were divided into blank control group and ginsenoside Rb1 group.RT-PCR was utilized to determine mRNA expressions of nerve growth factor (NGF),brain-derived neurotrophic factor (BDNF),glial derived neurotrophic factor(GDNF) and neural cell adhesion molecule (N-CAM) in the two groups.ELISA analysis was performed to measure secretion levels of NGF,BDNF and GDNF in the cultural supernatant.Results MTF analysis suggested ginsenoside Rb1 promoted proliferation of OECs with optimal effect at 20 μg/ml concentration for 48 hours (0.648±0.019,P ＜ 0.05).RT-PCR analysis demonstrated that mRNA expressions of NGF,BDNF,GDNF and N-CAM were significantly up-regulated in ginsenoside Rb1 group compared to those in blank control group (0.620 ± 0.011 vs 0.180 ± 0.011,0.511 ± 0.090 vs 0.293 ± 0.051,0.343 ± 0.042 vs 0.064 ± 0.005,0.839 ± 0.017 vs 0.717 ± 0.044) (P ＜ 0.05).ELISA analysis confirmed that secretions of NGF,BDNF and GDNF was increased in Rb1 group compared to those in blank control group (200.167 ± 8.361 vs 51.467 ± 3.815,156.700 ± 4.190 vs 96.500 ± 2.707,26.264 ± 5.864 vs 4.917 ± 10.894,P ＜ 0.05).Conclusion Ginsenoside Rb1 significantly promotes proliferation and bioactivity of OECs and hence benefits to spinal cord injury repair.