基于“血脉和利,精神乃居”探讨参麻益智方通过脑微血管内皮改善血管性痴呆大鼠突触可塑性的机制  

Exploration on the mechanism of Shenma Yizhi Formula in improving the synaptic plasticity of vascular dementia rats through cerebral microvascular endothelium based on the principle of‘harmonious blood vessel leads good spirit’

作　　者：张震 王志勇[1] 刘剑刚[1] 李浩 ZHANG Zhen;WANG Zhi-yong;LIU Jian-gang;LI Hao(Xiyuan Hospital of CACMS,Beijing 100091,China;Wangjing Hospital of CACMS,Beijing 100102,China)

机构地区：[1]中国中医科学院西苑医院,北京100091 [2]中国中医科学院望京医院,北京100102

出　　处：《中华中医药杂志》2023年第5期2317-2322,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：科技部国家重点研发计划“中医药现代化”重点专项(No.2022YFC3501400);国家中医药管理局中医药创新团队及人才支持计划项目(No.ZYYCXTD-C-202007);国家自然科学基金项目(No.82004434);中国中医科学院优秀青年科技人才培养专项(No.ZZ-14-YQ-001);中国中医科学院西苑医院国自然培育项目(No.XY20-08)。

摘　　要：目的:研究观察参麻益智方(SYF)改善血管性痴呆(VaD)模型大鼠学习记忆能力和对脑组织微血管内皮细胞(BMEC)和突触可塑性的机制。方法:SPF级雄性Wistar大鼠建立双侧永久性颈动脉结扎模型,手术成功的大鼠随机分为6组,每组10只,即模型组,SYF低、高剂量组(3.3、16.5 g·kg^(-1)·d~(-1)),血管内皮生长因子受体2(FLK-1)抑制剂Semaxinib (SU5416)组(25 mg/kg,腹腔注射)。另设假手术组(仅分离血管,穿线不结扎),模型组和SU5416组给予等容积蒸馏水灌胃,给药4周。3周末后使用水迷宫评估大鼠的学习记忆能力,麻醉后取材进行大脑海马病理观察,免疫组织化学染色法观察脑微血管内皮细胞,蛋白质印迹(Western Blot)和逆转录-聚合酶链反应(RT-PCR)检测海马组织的血管内皮生长因子(VEGF)、FLK-1、分裂原激活蛋白激酶(p38 MAPK)、N-甲基-D天冬氨酸受体1(NMDAR1)、突触后致密物95(PSD-95)的表达。结果:与模型组比较,SYF低、高剂量组均能显著缩短大鼠在水迷宫的潜伏期(P<0.05),显著增加穿越平台次数及在第一象限路程(P<0.05);显著增加海马组织的VEGF、FLK-1、p38 MAPK1、NMDAR、PSD-95的mRNA和蛋白表达水平(P<0.05)。结论:SYF可通过VEGF/FLK-1/p38MAPK信号通路,从BMEC调控突触可塑性进而对VaD模型大鼠起到保护作用,符合“血脉和利,精神乃居”理论。Objective:To study the effects of Shenma Yizhi Formula(SYF)on learning and memory ability,brain microvascular endothelial cells(BMEC)and synaptic plasticity in vascular dementia(VaD)rats induced by 2-vessel occlusion.Methods:Model rats(Wistar,SPF,male)were established by bilateral permanent carotid artery ligation.Model rats were randomly divided into 6 groups model group,SYF low-dose group(3.3 g·kg^(-1)·d^(-1)),SMY high-dose group(16.5 g·kg^(-1)·d^(-1)),and vascular endothelial growth factor receptor 2(FLK-1)inhibitor Semaxinib(SU5416)group(25 mg/kg,intraperitoneal injection),10 rats in each group:The sham operation group(only separate blood vessels,thread without ligation),model group and SU5416 group were given the same volume of distilled water by gavage for 4 weeks.At the end of three weeks,using the Morris water maze evaluated learning and memory ability of rats.After anesthesia,observed the pathology of hippocampus and the immunohistochemistry section of microvascular endothelial cells.Western blot and reverse transcription-polymerase chain reaction(RT-PCR)were used to observe the expression of glutamate vesicle transporte,vascular endothelial growth factor(VEGF),vascular endothelial growth factor receptor 2(FLK-1),mitogen activated protein kinase(p38 MAPK),N-methyl-D-aspartate receptor 1(NMDAR1)and postsynaptic density 95(PSD-95).Results:Compared with the model group,the latency of the rats in the water maze were significantly shortened(P<0.05).The times of platform crossing and the first quadrant distance were significantly increased(P<0.05)in the low-dose SYF group and the high-dose SYF group.The mRNA and protein expressions of VEGF,FLK-1,p38 MAPK,NMDAR1,PSD-95 in hippocampus were significantly increased(P<0.05).Conclusion:SYF can regulate synaptic plasticity from BMEC through VEGF/FLK-1/p38 MAPK signaling pathway,thus protect VaD model rats.The experimental results are in line with the theory of‘harmonious blood vessel leads good spirit’.

关 键 词：参麻益智方 血管性痴呆 络病 血管内皮生长因子 突触可塑性 

分 类 号：R285.5[医药卫生—中药学]