五脏温阳化瘀汤对PI3K/Akt-mTOR通路介导动脉粥样硬化型血管性痴呆自噬与凋亡的效应机制

Atherosclerosis (AS) vascular dementia (VaD) has brain dysfunction based on the pathogenesis of AS, it is also an acquired intelligence injury syndrome which mainly caused by the neuronal cell death in hippocampal CA1 area. The TCM fundamental pathogenesis of this disease is Deficiency-Yang and Excess-Yin. Extensive clinical practice and experimental studies confirmed that Wuzangwenyanghuayu Decoction had antioxidant effect and curative effect on AS-VaD. The latest research showed that the PI3K/Akt-mTOR pathway regulated the neuronal apoptosis and autophagy，which was closely related to the VaD. We Suppose that Wuzangwenyanghuayu Decoction maybe protect the nerve cell by adjusted the regulation of autophagy and apoptosis which mediated by PI3K/Akt-mTOR pathways in AS-VaD. Our project using copying AS-VaD rat model to observe the hippocampal CA1 neurons autophagy in electron microscopy; and then testing the expression of PI3K/Akt-mTOR pathway downstream autophagy and apoptosis associated protein Beclin-1, LC3, Caspase-9, Caspase-3 and Bcl-2 by immunohistochemistry, RT-PCR and Western Blot methods. The purpose of our project is to explore the effector mechanism of Wuzangwenyanghuayu Decoction treated on AS-VaD and provide a scientific basis for the TCM theory " Yang convert pneuma, Yin become entity".

动脉粥样硬化(AS)型血管性痴呆(VaD)是在AS的病理基础上出现脑功能障碍，以海马CA1区为主的神经细胞死亡产生获得性智能损害综合征。其中医病机本质是"阳化不足，阴化太过"。前期大量临床实践及实验研究证实五脏温阳化瘀汤治疗AS型VaD，疗效确切,且具有抗氧化作用。最新研究表明VaD与PI3K/Akt-mTOR通路调控神经细胞的凋亡和自噬关系密切。我们认为五脏温阳化瘀汤可能通过调节PI3K/Akt-mTOR通路介导AS型VaD自噬与凋亡而保护神经细胞。本项目通过复制AS型VaD大鼠模型，电镜观察海马CA1区神经细胞的自噬体；免疫组化、RT-PCR、Western Blot等检测PI3K/Akt-mTOR通路及其下游自噬与凋亡相关蛋白Beclin-1、LC3、Caspase-9、3及Bcl-2等表达，探讨五脏温阳化瘀汤干预AS型VaD的效应机制，为揭示中医"阳化气、阴成形"理论提供科学依据。

血管性痴呆(VaD)出现的脑功能障碍，以海马CA1 区为主的神经细胞死亡产生获得性智能损害综合征。其中医病机本质是“阳化不足，阴化太过”。前期大量临床实践及实验研究证实五脏温阳化瘀汤治疗VaD，疗效确切，且具有抗氧化作用。最新研究表明VaD 与PI3K/Akt-mTOR 通路调控神经细胞的凋亡和自噬关系密切。本项目在前期研究基础上，通过五脏温阳化瘀汤干预VaD 大鼠模型，电镜观察CA1 区神经细胞的自噬体，免疫组化观测大鼠海马区细胞形态，RT-PCR、Western Blot 等检测PI3K/Akt-mTOR 通路及其下游自噬与凋亡相关蛋白Beclin-1、LC3、Caspase-9、3 及Bcl-2 等表达，探讨五脏温阳化瘀汤干预AS 型VaD 的效应机制，研究发现，五脏温阳化瘀汤具有改善VaD 大鼠模型细胞形态，降低VaD 大鼠海马CA1自噬量，以及调节PI3K/Akt-mTOR 通路及相关因子表达的作用，且具有药物依赖性。由此认为，五脏温阳化瘀汤的抗VaD机制与调节PI3K/Akt-mTOR 通路及其下游自噬与凋亡有关，本研究结论为进一步丰富五脏温阳化瘀汤抗VaD机制提供了新的依据，为诠释中医“阳化不足，阴化太过”的科学内涵提供了新的基础。