摘要/Abstract

摘要： 目的 ·观察知母皂苷元（anemarrhena saponin，ZMS）对 H2O2损伤人神经母细胞瘤细胞 SH-SY5Y中脑源性神经营养因子（brain-derived neurotrophic factor，BDNF）的调控作用，并初步探索其分子机制。方法 ·用 H2O2损伤 SH-SY5Y细胞建立氧化应激细胞模型。通过 CCK-8试剂盒检测经 H2O2损伤后的 SH-SY5Y细胞活力。采用实时荧光定量 PCR（quantitative real-time PCR，qPCR）检测 BDNF及其重要转录子的 mRNA水平。利用组蛋白脱乙酰酶（histone deacetylases，HDACs）活性荧光分析试剂盒检测 ZMS对 HDACs活性的影响。采用蛋白质印迹（Western blotting）检测乙酰化组蛋白 H3、H4，特定乙酰化位点相关蛋白及 HDAC1/2/3蛋白的表达水平。结果 · qPCR检测显示， ZMS可以上调受损 SH-SY5Y细胞中 BDNF及转录子Ⅳ的 mRNA水平。 Western blotting检测显示，ZMS预处理后可增加乙酰化组蛋白 H3、H4、H3K14的蛋白水平，而对 HDAC1/2/3蛋白的表达无显著性影响。 HDACs活性荧光分析试剂盒检测结果显示， ZMS可抑制 HDACs的活性。结论 · ZMS可增加氧化应激细胞模型中 BDNF及转录子Ⅳ的 mRNA水平，其作用机制可能与调节组蛋白乙酰化水平相关。
关键词: 知母皂苷元, 氧化应激, 脑源性神经营养因子, 组蛋白乙酰化
Abstract:
Objective · To investigate the effect of anemarrhena saponin (ZMS) on mRNA level of brain-derived neurotrophic factor (BDNF) and relevant mechanism in oxidative stress damage of SH-SY5Y cells. Methods · SH-SY5Y cells treated with H2O2 were chosen as cell models of oxidative stress. Cell viability was determined using cell counting kit-8 (CCK-8). The mRNA levels of BDNF and its important transcripts were detectedquantitative real-time PCR (qPCR). The histone deacetylases (HDACs) activity fluorescence quantification assay kit was used to measure the effect of ZMS on HDACs activity. Western blotting was used to detect the protein levels of acetylated histone H3, acetylated histone H4, specific acetylation site-related proteins, and HDAC1/2/3. Results · qPCR showed that ZMS could increase the mRNA levels of BDNF and its transcript Ⅳ in the cell models. Western blotting showed that ZMS pretreatment could increase the protein levels of acetylated histone H3, acetylated histone H4 and acetylated histone H3K14, and there was no significant effect on protein levels of HDAC1/2/3. In addition, HDACs activity fluorescence quantification assay kit showed that ZMS could inhibit HDACs activity significantly. Conclusion · ZMS can increase the mRNA levels of BDNF and its transcript Ⅳ in oxidative stress damage cell models, which may be related to the regulation of histone acetylation level.
Key words: anemarrhena saponin (ZMS), oxidative stress, brain-derived neurotrophic factor (BDNF), histone acetylation


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