补阳还五汤载药血清对缺氧复氧神经干细胞SOD1蛋白及Nrf2mRNA的影响及机制的研究
首发时间:2012-11-22
摘要:目的:基于Nrf2调控水平探讨补阳还五汤载药血清保护缺氧复氧神经干细胞(neural stem cells, NSCs)的分子机制。方法:从14天胎鼠中获取胚胎神经干细胞。取传三代神经干细胞作为研究对象,随机分为:缺氧对照组、缺氧对照血清组(添加10%对照血清)、缺氧载药血清组(添加10%载药血清)三组,三气培养箱予以缺氧培养(氧浓度为5%)0.5h,再置于常规培养箱培养;实验中设定常规培养箱对照组。于复氧培养30min时,采用荧光定量PCR检测各组细胞中Nrf2mRNA转录状况;于复氧培养45min时间点采用免疫细胞化学法检测各组细胞SOD1表达;复氧培养1h时,分别测定各组细胞培养上清中MDA含量及SOD的抗氧化活性。结果:与缺氧对照组比较,经补阳还五汤载药血清干预实验组中Nrf2mRNA转录水平显著上调,同时观察到SOD1表达增强、SOD活性增高、MDA含量减低,均有统计学意义(P<0.01)结论:补阳还五汤载药血清在缺氧复氧早期通过有效上调神经干细胞Nrf2mRNA的转录水平,在一定时间内可有效减轻神经干细胞的氧化应激损伤。
关键词: 人体组织胚胎学 缺氧复氧 神经干细胞 Nrf2 补阳还五汤
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Effect and Mechanism of Serum Containing Drug of Buyang Huanwu Decoction on SOD1proteins and Nrf2mRNA in NSCs undergoing hypoxia-reoxygenation
Abstract:Objective: TO explore the the protective mechanism based on Nrf2 of Serum Containing Drug of Buyang Huanwu Decoction on neural stem cells injured by hypoxia-reoxygenation. Methods: Neural stem cells from 14 days fetal rat were cultured in 5%O2 for 0.5h, adding 10% serum Containing Drug of Buyang Huanwu Decoction ,t and then placing them in 21%O2 incubator . At the same time, experimental control group(adding 10% control serum) 、model set and normal control group were set respectively. Nrf2mRNA of NSCs were detected after 30min undergoing reoxygenation by fluorescence quantitative PCR; SOD1 expression after 45min undergoing reoxygenation using reaeration immunocytochemical assay; the content of MDA and the antioxidant activity of the SOD in the culture supernatant after 60min undergoing reoxygenation by colorimetric method respectively.Results: The level of Nrf2mRNA were raised significantly, and SOD1 expression was enhanced obviously, and SOD activity increased, meanwhile MDA content reduced (P<0.01) in hypoxia drug-serum group.Conclusions: Serum Containing Drug of Buyang Huanwu Decoction can protect NSCs from hypoxia-reoxygenation injure by raising the transcription level of Nrf2mRNA in early stage.
Keywords: Human histology and embryology hypoxia-reoxygenation neural stem cells Nrf2 buyang huanwu decoction
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补阳还五汤载药血清对缺氧复氧神经干细胞SOD1蛋白及Nrf2mRNA的影响及机制的研究
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