基于calpain1-calcineurin-TFEB通路介导的神经元ALP障碍研究西洋参拟人参皂苷F11抗脑缺血作用机制

拟人参皂苷F11（Pseudoginsenoside F11, PF11）是西洋参根、茎叶中的奥克梯隆型三萜皂苷，是其抗脑缺血作用的有效成分之一。我们发现，PF11抑制脑缺血后神经元calpain1（CAPN1）的激活，且以calcineurin（CN）依赖的方式促进TFEB入核、缓解自噬-溶酶体通路（ALP）阻滞，抑制缺血损伤。结合相关文献报道，我们提出“脑缺血后神经元存在CAPN1-CN-TFEB介导的ALP阻滞机制，而PF11通过抑制神经元CAPN1的激活，改变CN的功能而促进TFEB入核，进而缓解ALP障碍，最终发挥抗脑缺血作用”这一假说，运用Nestin-Cre:LSL-Cas9-EGFP双转基因小鼠及皮层神经元结合病毒介导目的基因敲除技术，采用在体永久性脑缺血和皮层神经元氧糖剥夺模型，阐明PF11调控神经元CAPN1-CN-TFEB介导ALP障碍发挥抗脑缺血的作用机制。

As one of active ingredient isolated from the root, stem and leaf of Panax quinquefolium L., Pseudoginsenoside F11 (PF11) belongs to ocotillol-type saponins and exerts protective effects on cerebral ischemic injury. We previously reported that PF11 significantly suppressed the activation of calpain1 (CAPN1), promoted TFEB nuclei translocation and alleviated the impairment of autophagy-lysosomal pathway (ALP) in neurons, which could be reversed by the inhibitor of calcineurin (CN). Thus, in the present study, we will use the Nestin-Cre: LSL-Cas9-EGFP double-transgenic mice and primary cultured neurons to investigate the mechanisms of CAPN1-CN-TFEB-mediated impairment of ALP in neurons during ischemic injury induced by permanent middle cerebral artery occlusion or oxygen glucose deprivation. Moreover, we will further explore the autophagy-related mechanisms of PF11 on alleviating the ischemic injury.