基于MEF2-PGC1α信号通路探讨药根碱对线粒体的保护作用及机制

The dysfunction of mitochondira induced by Aβ, oxidative stress and the environment plays a critical role in the progression of AD. We have shown that Jatrorrhizine act on anti-oxidative stress protect mitochondrial function. PGC1α is a main factor in regulating biogenesis and function of mitochondria. Activation of MEF2 results in the transcription of MEF2 and PGC1α, MEF2-PGC1α signaling pathway could protect mitochondrial function and inhibit neuronal apoptosis. By use of Jatrorrhizine treatment N2a/APP695 cell mode and APP/PS1 dtg mice animal mode, Based on previous work by Western blot, TUNEL and co-immunoprecipitation and other methods, this project is to investigate the underlying mechanisms of Jatrorrhizine for anti-oxidative stress and mitochondrial pathway on apoptosis by testing changes in protein expression levels in Keap1/Nrf2/ARE, MAPK and PI3K/Akt/mTOR signaling pathway. observed protein expression levels in MEF2-PGC1α signaling pathway, Association in ROS and PGC1α, MEF2 and PGC1α, and focus on modulation of MEF2-PGC1α transcriptional network, to further clarify the molecular mechanism for protective role of Jatrorrhizine on neuronal injury in AD .

线粒体功能障碍在AD的发生发展过程中起着关键作用。我们前期研究已发现药根碱具有抗氧化应激和保护线粒体功能。PGC1α是调节线粒体生物合成及功能的核心因子，MEF2的活化可激活PGC1α的转录，MEF2-PGC1α信号通路能保护线粒体功能并抑制神经元凋亡。本项目在前期工作的基础上拟应用N2a/APP695细胞和APP/PS1转基因小鼠探讨药根碱对线粒体的保护作用及机制。采用Western blot、TUNEL、PCR及免疫共沉淀等观察Keap1/Nrf2/ARE、MAPK与PI3K/Akt/mTOR等信号通路蛋白表达水平变化，完善药根碱抗氧化应激及抑制线粒体途径凋亡的分子机制；观察MEF2-PGC1α信号通路蛋白表达水平变化及ROS与PGC1α、MEF2与PGC1α的关联，重点探讨药根碱对MEF2-PGC1α转录网络的调控作用，进一步明确药根碱在AD神经元损伤中的保护作用及其分子机制。

线粒体功能障碍在AD的发生发展过程中起着关键作用。我们前期研究已发现药根碱具有抗氧化应激和保护线粒体功能。PGC1α是调节线粒体生物合成及功能的核心因子，MEF2的活化可激活PGC1α的转录，MEF2-PGC1α信号通路能保护线粒体功能并抑制神经元凋亡。本项目在前期工作的基础上拟应用N2a/APP695细胞和APP/PS1转基因小鼠探讨药根碱对线粒体的保护作用及机制。采用Western blot、TUNEL、PCR及免疫共沉淀等观察Keap1/Nrf2/ARE、MAPK与PI3K/Akt/mTOR等信号通路蛋白表达变化，完善药根碱抗氧化应激及抑制线粒体途径凋亡的分子机制；观察MEF2-PGC1α信号通路蛋白表达水平变化及ROS与PGC1α、MEF2与PGC1α的关联，重点探讨药根碱对MEF2-PGC1α转录网络的调控作用，进一步明确药根碱在AD神经元损伤中的保护作用及其分子机制。本项目的各项研究表明，药根碱具有抗氧化应激作用，并通过Keap1/Nrf2/ARE信号通路发挥抗氧化应激作用；药根碱对AD线粒体具有保护作用，并通过影响MAPK及PI3K/Akt/mTOR信号通路发挥抗凋亡作用；药根碱对AD细胞及动物模型MEF2-PGC1α转录网络具有调控作用；药根碱能够明显改善AD动物模型的学习记忆能力；药根碱对AD动物模型Aβ代谢的调节作用。本项目的的各项研究进一步明确了药根碱对神经元损伤的保护作用及其分子机制，为其研发成一种有效防治AD药物提供了理论依据。