中药山茱萸及其有效成分莫诺苷对脑卒中后神经血管稳态重构有效治疗时间窗的研究

The treatment time-window of stroke is short, and less than 1% of stroke patients can take thrombolytic therapy in China. .Based on the hypothesis of “neurovascular homeostasis rehabilitation”, we use MCAO rat model to study the therapeutic time window of morroniside on ischemic stroke, which is isolated from Chinese traditional medicine Cornus officinalis. Morroniside treatment begin at different times including 3 h, 6 h, 12h, 24 h, 3d, or 7d. In the time-window study, ① behavioural recovery will be evaluated at repeated time intervals on locomotor activity, grip strength, ability to remain on an accelerating rotarod, sensory neglect, gait impairment; ② TTC staining and MRI testing are used to examine the infarction volume; ③ the permeability of BBB is analyzed; ④ immunofluorescence staining of BrdU+、BrdU+/DCX+、BrdU+/NeuN+ cells are used for analysis neurogenesis; ⑤ immunofluorescence staining of BrdU+/laminin+、CD105+ vessels are used for analysis neurogenesis; ⑥ western-blot method is used to detect neurotrophic factors such as NGF、BDNF; ⑦ signaling pathways involved in neurogenesis and angiogenesis are analyzed..The present study will establish a new model of time-window study based on neurovascular homeostasis rehabilitation, and determine morroniside’s time-response effects on infarct size, behavioural performance and neurorestoration on a variety of clinically relevant tests given after MCAO. Large time-window of treatment including morroniside is of a great significance to reduce disability and mortality of stroke.

脑卒中治疗时间窗很短，我国不足1%病人能接受溶栓治疗。. 在“神经血管稳态重构”假说基础上，大鼠模型不同时间点灌服从山茱萸筛选的莫诺苷， ①研究大鼠自主活动能力、感觉忽视、步态分析等对神经功能影响，②核磁共振、TTC染色等检测脑梗死体积对神经影像学影响，③血脑屏障通透性、MMP2、TNF-α、IL-1β等对血脑屏障重构影响，④免疫荧光检测BrdU、BrdU+/DCX+、BrdU+/NeuN+细胞数量等对神经 再生影响，⑤免疫荧光检测BrdU+/laminin+、CD105阳性新生血管数量等对血管新生影响，⑥免疫印迹检测NGF、BDNF等神经血管再生相关因子影响，⑦免疫印迹检测Wnt、β-catenin、EphB4等对Wnt和EphB4信号通路影响。.探索脑卒中治疗时间窗研究方法和神经稳态重构影响因素，确定莫诺苷时间窗，争取90%病人能接受治疗，对降低脑卒中死亡率和致残率有重要意义。

脑卒中治疗时间窗很短，我国不足1%病人能接受溶栓治疗。在该基金的支持下，我们采用大鼠大脑中动脉阻塞模型进行莫诺苷有效治疗时间窗的研究。从神经功能、活体或者离体神经影像学、血脑屏障重构、神经再生、血管新生、神经因子、干细胞调节信号通路等7个方面对MCAO造模后3h、6h、12h、24h、48h、72h几个时间点灌胃给予莫诺苷的药效学进行了评价，建立起整体动物、组织器官、细胞和分子水平交叉验证有效时间窗的研究方法，明确了莫诺苷促进神经稳态重构的有效治疗时间窗，为今后用药指导提供坚实的实验基础，期待后续研究得到转化，最终为脑卒中的临床康复治疗提供有效的方法。在基金资助下，培养硕士博士研究生共计12人；发表SCI论文5篇，中文核心期刊论文12篇；获批专利授权1项；相关研究数据申报北京市实验动物行业科学进步奖，获得一等奖。