黄芩及黄芩苷基于Rho/ROCK调节线粒体动态改善糖尿病脑病的作用及机制研究

Developing drugs that regulate mitochondrial dynamics and protect mitochondrial functions has become the hotspot of recent researches on the treatment of diabetic encephalopathy. Rho/ROCK plays an important role in the regulation of mitochondrial fusion and fission dynamics, whose activation can lead to an imbalance of mitochondrial dynamics, resulting in neuronal synaptic damage and cell pyroptosis. Scutellariae Radix is a widely used traditional Chinese medicine for the treatment of neurological diseases, while its active ingredient baicalin has also been reported to have a significant neuroprotective effect. Our previous studies showed that Scutellariae Radix and baicalin significantly improved the neuron pyroptosis in the hippocampus of diabetic mice and alleviated cognitive dysfunction caused by metabolic disorders. However, the molecular mechanism underlying its beneficial effects is not clear. Meanwhile, as an important indicator of quality control in Scutellariae Radix, the pharmacological contribution of baicalin in improving diabetic encephalopathy is still unknown. Under the guidance of traditional Chinese medicine theories, this project will apply the gene knockdown/over-expression technology with recombinant lentivirus and the molecular biology technologies to explore the role of Rho/ROCK pathway on mitochondrial dynamics, and to verify the regulation target of baicalin and the underlying mechanisms. On this basis, by knockout of baicalin from Scutellariae Radix, the contribution of baicalin to the pharmacological effects of Scutellariae Radix is also to be clarified. Therefore, this project will help us understand the effects of baicalin on improving diabetic encephalopathy and its pharmacological contribution towards Scutellariae Radix, which will bring us a lot of benefits on the material basis and scientific meanings of traditional Chinese medicine.

调节神经细胞线粒体动态平衡，开发具有线粒体保护功能的药物，是近年糖尿病脑病研究的热点之一。Rho/ROCK是调节线粒体融合和裂分动态的重要途径，该通路的激活会引发线粒体动态失衡，造成突触损伤和神经细胞焦亡。黄芩是糖尿病脑病治疗的常用中药，黄芩苷是其神经保护作用的主要活性成分。前期研究表明，黄芩和黄芩苷能改善糖尿病脑病小鼠的认知功能障碍，减少代谢紊乱引起的神经细胞焦亡，但具体机制尚不清楚；同时，黄芩苷作为黄芩质量控制指标，在黄芩改善糖尿病脑病药效中的贡献仍未明了。本项目拟在中医药理论指导下，采用重组慢病毒构建的基因原位敲减/过表达动物模型以及相关分子生物学技术，重点研究Rho/ROCK通路与神经细胞线粒体动态作用关系，从而阐明黄芩苷缓解细胞焦亡的关键调控因子以及改善糖尿病脑病的分子机制。在此基础上从黄芩中敲除黄芩苷，进一步探明黄芩苷对黄芩药理功效的贡献，从而揭示中药作用的物质基础和科学内涵。

通过本项目的实施，阐明了黄芩及黄芩苷调节神经细胞线粒体融合裂分动态平衡，减少神经细胞焦亡，改善T2DE引发认知障碍的分子机制。项目研究了黄芩苷对线粒体功能的保护作用，以及对线粒体融合裂分动态平衡的调节，明确了Rho/ROCK是黄芩苷调节线粒体裂分的重要作用通路；同时，研究阐明了黄芩苷可通过PI3K/AKT/FoxO1通路抑制小胶质细胞的过度激活，作用于钠氢交换因子1、钙蛋白酶1调节相关阳离子水平，抑制炎症小体NLRP3调节的TGF-β1/NF-κB炎症级联反应，改善脑内Cdk5/IRS/Tau胰岛素通路，缓解神经细胞焦亡，从而改善T2DE导致认知障碍的作用机制。在此基础上采用中药成分敲除技术探究黄芩发挥神经保护的药效物质基础，黄芩苷是黄芩的质量控制指标，本项目通过研究黄芩苷和黄芩苷阴性样品在黄芩神经保护药理作用中的贡献，阐明黄芩发挥神经保护的药效物质基础，为黄芩苷作为黄芩质量控制指标提供科学依据。综上，本项目在中医药理论指导下，运用现代中药药理相关技术，深入研究中药黄芩及黄芩苷改善T2DE引发认知障碍的分子作用和药效物质基础，为黄芩的临床使用提供了翔实的理论支撑和实验依据，相关方法和数据为进一步研究和开发中药新药奠定了基础。