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刘娟,裘莹,何晶,等.人参皂苷Rb1和Rg1对海马神经元的影响[J].同济大学学报（医学版）,2011,32(3):1-6. [点击复制]
LIU Juan,QIU Ying,HE Jing,et al.Effects of ginsenoside Rb1 and Rg1 on hippocampal neurons[J].同济大学学报（医学版）,2011,32(3):1-6. [点击复制]

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人参皂苷Rb1和Rg1对海马神经元的影响
刘娟¹,裘莹²,何晶¹,文蕾³,管晓菲³,袁琼兰¹
0 字体:加大+\|默认\|缩小-
(1.同济大学医学院解剖学与神经生物学教研室,上海200092;2.同济大学医学院病理解剖学教汧室，上海20092;3.同济大学2006级临床医学八年制学生,上海20092)

摘要:

目的探讨人参皂苷（ginsenoside）Rb1和Rg1（GRb1，GRg1）对海马神经元突起生长及神经保护作用及其机制。方法培养SD新生大鼠海马神经元，分为：正常组、Rb1组、Rg1组、Rb1＋API-2（Akt抑制剂）、Rg1＋API-2、Rb1＋PD98059（MEK抑制剂）和Rg1＋PD98059组，培养1d，用免疫染色观察神经突起的生长。加入Aβ25-35制备海马神经元损伤模型，分为正常组、损伤组（AB组）、Rb1组、Rg1组、Rb1＋API-2、Rg1-I-API-2、Rb1＋PD98059和Rg1＋PD98059，培养48h，用Hoechst33258染色观察活细胞与凋亡细胞。用Elisa观察培养上清液里神经营养因子的浓度（nerve growth factor，NGF；brain—derived neurotrophic factor，BDNF；neurotrophin-3，NT-3）。结果 Rb1和Rg1组神经突起生长高于对照组（P＜0.05）；API-2和PD98059显著抑制了Rb1和Rg1诱导的神经突起生长（P＜0．01），API-2的抑制效果强于PD98059；Rg1＋API-2组BDNF高于对照组和Rg1组（P＜0.05），其余各组间差异无统计学意义（P＞0.05）；Rg1组NT-3的浓度高于对照组（P＜0.05）；Rb1＋PD98059组的NGF高于Rb1组。Rb1和Rg1组海马神经元的凋亡率显著低于损伤组（P＜0.01）；PD98059和API-2阻断了Rb1和Rg1对神经元的保护作用（P＜0.01）；神经营养因子水平各组间差异无统计学意义。结论 Rb1和Rg1促进海马神经元突起生长及抵抗A1325-35损伤，促进神经元的存活。其机制与Akt和ERK1/2的信号通路激活有关，与增加神经营养因子的分泌无关。

关键词: 人参皂苷Rb1和Rg1 海马神经元神经突起 PD98059 API-2

DOI：10.3969/j.issn1008-0392.2011.03.001

基金项目:国家自然科学基金(30971531)

Effects of ginsenoside Rb1 and Rg1 on hippocampal neurons

LIU Juan¹,QIU Ying²,HE Jing¹,WEN Lei³,GUAN Xiao-fei³,YUAN Qiong-lan¹

(1.Dept.of Anatomy and Neurobiology,School of Medicine,Tongji University,Shanghai 20092,China;2.Dept.of Pathology,School of Medicine,Tongji University,Shanghai 20092,China;3.Eight Years of Clinical Medicine,School of Medicine,Tongji University,Shanghai 200092 China)

Abstract:

Objective To investigate the effects of ginsenoside Rb1 and Rg1(Rb1,Rg1) on neurite outgrowth of hippocampal neurons and protection hippocampal neurons against injury caused by Aβ25- 35.Methods Cultured hippocampal neurons from newborn SD rats were divided into groups; normal,Rb1,Rg1,Rb1+API-2(Akt inhibitor),Rg1+API-2,Rb1+PD98059(MEK inhibitor), and Rg1+PD98059.After 24 h of culture the neurite outgrowth was evaluated by immunostaining of neuronal growth associated protein-43(GAP-43).Hippocampal neurons were treated with Aβ25-35 and divided into groups;injured Aβgroup,Rb1,Rg1,Rb1+API-2,Rg1+API-2,Rb1+PD98059 and Rg1+PD98059.After 48 h of cultured the Hoechst33258 staining was performed to evaluate the rate of apoptosis.In addition,the nerve growth factor(NGF),brain-derived neurotrophic factor (BDNF) and neurotrophin-3(NT-3) were analyzed by ELISA method.Results The neurites were significantly longer and the number increased in Rb1 and Rg1 groups compared with normal group(P＜0.05).API-2 and PD98059 blocked neurite outgrowth significantly(P＜0.01),and API-2 was more efficient than PD98059.No significant difference of BDNF from supernatant was observed between groups except that BDNF in Rg1+API-2 group was higher compared with Rg1 group;NT-3 in Rg1 group was significantly higher than that in normal group;NGF in Rb1 + PD98059 was significantly higher than that in Rb1 group(P＜0.05).Apoptotic rates of hippocampal neurons in Rb1 and Rg1 groups decreased significantly(P＜0.01) compared with Aβgroup.Moreover,API-2 and PD98059 significantly inhibited neuroprotection of Rb1 and Rg1 against Aβ25-35 injury(P＜0.05).No differences of NGF,NT-3 and BDNF were observed among groups(P＞0.05).Conclusion Ginsenosides Rb1 and Rg1 can induce neurite outgrowth of hippocampal neurons and protect neurons against Aβ25 - 35 injury.Their mechanisms are related to Akt and ERK1/2 signal transduction pathways but not to the increase of neurotrophins secretion.

Key words: ginsenoside Rbl and Rgl hippocampal neurons neurite outgrowth PD98059 API-2