脑通汤含药血清对缺氧/复氧大鼠海马神经元抗氧化应激相关指标的影响  

Effect of Naotong soup medicinal serum on anti-oxidative stress-related indicators of hippocampal neurons in rats after hypoxia /reoxygenation

作　　者：何筑 况时祥[2] 张树森[2] 邹家莉[2] 杨燕[3] 崔冬冰[3] 

机构地区：[1]贵阳市第三人民医院,贵州贵阳550002 [2]贵阳中医学院第二附属医院 [3]贵阳医学院组织工程与干细胞实验中心

出　　处：《中国老年学杂志》2016年第3期526-529,共4页Chinese Journal of Gerontology

基　　金：国家自然科学基金资助项目(No.81160490)

摘　　要：目的探讨脑通汤通过海马神经元抗氧化应激治疗血管性痴呆(VD)的机制。方法培养大鼠原代海马神经元及鉴定,取培养9 d的神经元,随机分为正常细胞组、缺氧/复氧模型组、正常血清组、脑通汤大、中、小剂量含药血清组。除正常细胞组以外,各组均置入(95%N2+5%CO2)混合气体的三气培养箱缺氧24 h再复氧2 h造模。采用MTT法检测缺氧24 h和复氧2 h时间点海马神经元活性并计算细胞存活率;实时荧光定量PCR法检测海马神经元核因子-E2-相关因子(Nrf)2、血红素氧合酶亚型(HO)-1、醌氧化还原酶(NQO)1 mRNA的表达。结果 MTT显示:缺氧24 h:模型组的海马神经元活性及存活率较正常细胞组明显下降(P<0.05),与正常血清组及脑通汤不同剂量含药血清组比较无明显差异(P>0.05)。复氧2 h:脑通汤不同剂量含药血清组均较模型组明显升高(P<0.05),正常血清组较模型组无明显差异(P>0.05)。实时荧光定量PCR结果显示:脑通汤含药血清能够促进缺氧/复氧海马神经元抗氧化应激相关基因Nrf2、HO-1、NQO1的表达(P<0.05),并且脑通汤含药血清大剂量组与中剂量组的上述3个基因水平明显高于缺氧/复氧模型组(P<0.01)。结论脑通汤可通过上调抗氧化相关基因Nrf2、HO-1、NQO1的表达,减轻海马神经元缺氧/复氧损伤的氧化应激反应,从而保护大鼠海马神经元缺氧/复氧损伤。Objective To investigate Naotong soup treated vascular dementia by hippocampal neurons against oxidative stress. Methods Primary hippocampal neurons were cultured and indentified. Neurons cultured for 9 days were randomly divided into normal cell, hypoxia/reoxygenation（H/R） model, normal serum, large, medium and small doses of Naotong soup medicinal serum groups. The activity and the survival rate of neurons both at 24 hours after hypoxia and 2 hours after reoxygenation were evaluated by colorimetric MTT assay. Real-time fluorescent quantitative PCR was used to detect the expressions of nuclear factor erythroid-derived factor 2-related factor（ Nrf2 ）, Heme Oxygenase（HO） -1 and NAD（P） H ：quinone oxidoreductase（ NQO1 ） mRNA. Results Compared with those of normal cell group, hippocampal neuronal activity and the survival rate of model groups were significantly lower after hypoxia 24 h（ P〈0.05 ）, between normal serum group and different dose of Naotong soup medicinal serum groups, they had no significant differences （ P〉0.05 ）. Compared with those of model group, they had significantly increased after reoxygenation 2 h in different dose of Naotong soup medicinal serum groups（ P〈0.05 ）and in nor- mal serum group they had no significant difference （ P〉0.05 ）. Naotong soup medicinal serum promoted the expressions of Nrf2, HO-1, NQO1 of hippocampal neurons（ P〈0.05 ）, and these three genes were significantly higher than those of model group（ P〈0.05 ）. Conclusions Naotong soup could increase the expressions of Nrf2, HOwl and NQO1 and reduce oxidative stress of hippocampal neurons after H/R. So it could protect hippocampal neurons in rats after H/R.

关 键 词：脑通汤 海马神经元 缺氧/复氧 抗氧化应激 

分 类 号：R74[医药卫生—神经病学与精神病学]