基于肠-脑轴介导的LPS-TLR4/NLRP3交互串联通路探讨益智清心方改善阿尔茨海默病神经炎症的机制

肠道微生物失调通过肠-脑轴诱发外周和中枢神经炎症，导致阿尔茨海默病（Alzheimer's disease，AD），而LPS-TLR4/NLRP3交互串联通路是激活神经炎症的关键环节。益智清心方是针对AD“虚瘀-浊毒”中医病机制定的临床协定方，课题组前期研究发现，该方不仅能调节肠道微生态，还能通过肠-脑轴改善AD神经炎症，而LPS介导的炎症通路是其最主要的生物途径。故本项目提出科学假说：益智清心方通过调控肠-脑轴介导的LPS-TLR4/NLRP3交互串联通路，抑制神经炎症和细胞凋亡，改善AD认知功能。为验证该假说，本研究体内以APP/PS1双转基因小鼠为研究对象，体外采用Aβ诱导BV-2和PC12细胞共培养模型，充分利用16SrDNA及宏基因组测序、免疫印迹、免疫荧光等多种分子生物学技术，观察益智清心方对肠-脑轴介导的LPS-TLR4/NLRP3通路的影响，能够为AD的中医药防治提供依据。

Intestinal microbiome dysregulation induces peripheral and central neuroinflammation through the gut-brain axis, leading to Alzheimer's disease (AD), and LPS-TLR4/NLRP3 interactive tandem pathway is a key link in activating neuroinflammation. Yizhi Qingxin formula is a clinical protocol formulated for the pathogenesis of "deficiency of blood stasis and turbid toxin" of AD. Previous studies have found that this formula not only regulate intestinal microecology, but also improve AD neuroinflammation through the gut-brain axis, and LPS-mediated inflammatory pathway is the most important biological pathway. Therefore, we proposed a scientific hypothesis: Yizhi Qingxin formula can regulate LPS-TLR4/NLRP3 interactive tandem pathway mediated by the gut-brain axis, inhibit neuroinflammation and apoptosis, and improve the cognitive function of AD. In order to verify this hypothesis, this study will adopt APP/PS1 double transgenic mice in vivo, and LPS-induced BV-2 and PC12 cells in vitro, fully utilize 16SrDNA, metagomic sequencing integration, western blot and immunofluorescence, etc., to observe the effect of Yizhi Qingxin formula on LPS-TLR4/NLRP3 pathway mediated by the gut-brain axis, which can provide basis for the prevention and treatment of AD with traditional Chinese medicine.