[1]陆叶,袁媛,李双阳,等.通窍益智颗粒通过Ca2+/CaMK Ⅱ信号通路改善血管性痴呆大鼠海马CA1区突触可塑性机制研究[J].西部中医药,2025,38(06):22-28.[doi:10.12174/j.issn.2096-9600.2025.06.05]
　LU Ye,YUAN Yuan,LI Shuangyang,et al.Study on the Mechanism of Synaptic Plasticity in the Hippocampal CAI Region of Vascular Dementia Rats by Tongqiao Yizhi Granules via Ca2+/CaMKⅡ Signaling Pathway[J].Western Journal of Traditional Chinese Medicine,2025,38(06):22-28.[doi:10.12174/j.issn.2096-9600.2025.06.05]
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通窍益智颗粒通过Ca2+/CaMK Ⅱ信号通路改善血管性痴呆大鼠海马CA1区突触可塑性机制研究()

《西部中医药》[ISSN:2096-9600/CN:62-1204/R]

卷:
38
期数:
2025年06期
页码:
22-28
栏目:
基础研究
出版日期:
2025-06-15

文章信息/Info

Title:
Study on the Mechanism of Synaptic Plasticity in the Hippocampal CAI Region of Vascular Dementia Rats by Tongqiao Yizhi Granules via Ca2+/CaMKⅡ Signaling Pathway
作者:
陆叶, 袁媛, 李双阳, 王饶琼, 唐红梅, 白雪
西南医科大学附属中医医院，四川 泸州 646000
Author(s):
LU Ye, YUAN Yuan, LI Shuangyang, WANG Raoqiong, TANG Hongmei, BAI Xue
关键词:
血管性痴呆; 玄府; 钙离子; 磷酸化钙调蛋白依赖性蛋白激酶; 突触可塑性; 通窍益智颗粒
Keywords:
vascular dementia; sweating pore; Ca; phosphorylated calcium modulated protein-dependent protein kinase; synaptic plasticity; granules
分类号:
R256.29
DOI:
10.12174/j.issn.2096-9600.2025.06.05
文献标志码:
A
摘要:
目的探讨通窍益智颗粒对血管性痴呆（vascular dementia，VaD）大鼠的相关作用机制。 方法选取92只SD大鼠，其中12只纳入假手术组，其余80只大鼠采用双侧颈总动脉永久结扎法制备VaD大鼠模型，筛选出62只造模成功的SD大鼠，按随机数字表法分为模型组、美金刚组及通窍益智颗粒高、中剂量组，每组10只。假手术组、模型组予生理盐水灌胃；美金刚组大鼠以1.04 mg/（kg·d）剂量美金刚灌胃；通窍益智颗粒高、中剂量组分别以9.68、4.84 g/（kg·d）剂量通窍益智颗粒灌胃，每日灌胃2次，连续干预30天。采用Morris水迷宫方法评价大鼠行为学变化情况；透射电子显微镜观察大鼠海马CA1区神经元突触结构；流式细胞术检测大鼠海马神经元中Ca2+荧光强度；免疫荧光法测定N-甲基-D-天冬氨酸（N-methyl-D-aspartic acid receptor，NMDA）受体亚基Glu N1、Glu N2B以及α-氨基-3-羟基-5-甲基-4-异恶唑丙酸（α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid，AMPA）受体功能亚基Glu A2海马CA1区定位表达；蛋白免疫印迹法检测Glu N1、Glu N2B、Glu A2及钙调蛋白依赖性蛋白激酶（Calmodulin-dependent protein kinase，CaMKⅡ）和磷酸化钙调蛋白依赖性蛋白激酶（phosphorylation calmodulin-dependent protein kinase，p CaMKⅡ）蛋白表达情况。 结果与假手术组比较，模型组大鼠在目标象限游泳时间缩短，穿越原平台次数减少，神经元中Ca2+浓度升高，海马组织Glu N1、Glu N2B、Glu A2和p CaMK Ⅱ表达水平降低（P＜0.05）。实验第5天模型组大鼠EL时间延长（P＜0.05）；与模型组比较，美金刚组及通窍益智颗粒高、中剂量组大鼠在目标象限游泳时间延长，穿越原平台次数增多，神经元中Ca2+浓度均降低，海马组织Glu N1、Glu N2B、Glu A2和p CaMK Ⅱ表达水平升高（P＜0.05）。实验第5天美金刚组及通窍益智颗粒高、中剂量组大鼠EL时间缩短（P＜0.05）；与美金刚组、通窍益智颗粒高、中剂量组比较，通窍益智颗粒高剂量组各项指标改善最明显（P＜0.05）。模型组大鼠海马CA1区突触间隙变模糊，突触前后膜肿胀，结构难以区分，突触活性区变小，细胞内线粒体结构破坏；通窍益智颗粒高、中剂量组和美金刚组大鼠突触结构改善，突触前后膜结构较完整，突触间隙较清晰，突触小泡增多，细胞内可见完整线粒体，以通窍益智颗粒高剂量组改善最佳。 结论基于“玄府理论”组成的通窍益智颗粒能够改善VaD模型大鼠学习记忆能力及海马CA1区神经元突触结构，其潜在机制可能与其调节Ca2+/CaMK Ⅱ信号通路，改善突触可塑性有关。
Abstract:
ObjectiveTo explore the relevant mechanism of Tongqiao Yizhi (orifice-unobstructing wisdom-benefiting) granules in the treatment of vascular dementia (VaD). MethodsA total of 92 SD rats were selected, among them, 12 rats were included as sham operation group, and the remaining 80 rats were prepared into VaD models using bilateral permanent ligation of the common carotid artery, 62 rats, after successfully modeling, were screened, and divided into the model group, memantine group, high and moderate doses groups of Tongqiao Yizhi granules according to random number table method with ten rats in each group. Sham operation group and the model group were drenched with physiological saline; memantine group accepted intragastric administration of memantine, 1.04 mg/(kg·d); high and moderate doses groups of Tongqiao Yizhi granules were given the granules, 9.68 and 4.84 g/(kg·d), for gastric lavage respectively, twice each day, for 30 days continuously. Morris water maze was adopted to assess behavioral changes in rats; transmission electron microscopy was used to observe synaptic structure of the neuron in the hippocampal CAI region of the rats; flow cytometry was applied to detect fluorescence intensity of Ca2+ in hippocampal neurons of the rats; immunofluorescence was utilized to measure the expressions of NMDA receptor subunit Glu N1, Glu N2B and AMPA receptor subunit Glu A2 in hippocampal CAI region; Western blot was used to check the expressions of Glu N1, Glu N2B, Glu A2 and CaMKⅡ, and p CaMKⅡ protein. ResultsCompared with sham operation group, the time spent swimming shortened in the target quadrant and the number of platform crossings decreased, while the concentrations of Ca2+ in the neurons increased in the model group, the expressions of Glu N1, Glu N2B and Glu A2, and p CaMKⅡ protein lowered in hippocampal CAI region (P<0.05). EL time prolonged in the model group on the fifth day of the experiment (P<0.05); compared with the model group, the swimming time spent in the target quadrant prolonged and the number of platform crossings increased, the density of Ca2+ in the neurons decreased, the expressions of Glu N1, Glu N2B and Glu A2, and p CaMKⅡ raised in hippocampal CAI region in memantine group, high and moderate dose groups of Tongqiao Yizhi granules (P<0.05). EL time shortened in memantine group, high and moderate dose groups of Tongqiao Yizhi granules on the fifth day of the experiment (P<0.05). The improvements of the indexes were more significant in high dose group of Tongqiao Yizhi granules when compared with memantine group and moderate dose group of Tongqiao Yizhi granules (P<0.05). The synaptic cleft in CA1 region of rat hippocampus was blurred, the presynaptic and postsynaptic membranes were swollen, the structure was difficult to distinguish, the synaptic active area was reduced, and mitochondrial structure within the cells was destroyed in the rats of the model group; Synaptic structure was improved in memantine group, high and moderate dose groups of Tongqiao Yizhi granules, synaptic membrane structure was relatively complete, synaptic cleft was more clear, synaptic vesicles were increased in memantine group, high and moderate dose groups of Tongqiao Yizhi granules, complete mitochondria could be seen within the cells, and the improvements of high dose group of the granules were the best. Conclusion Tongqiao Yizhi granules based on "sweating pore theory" could improve learning and memory ability in VaD rat models, and synaptic structure of the neurons in hippocampal CAI region, and its potential mechanism might be related to that it could regulate Ca2+/CaMKⅡ signaling pathways and improve synaptic plasticity.

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备注/Memo

备注/Memo:
陆叶（1995—），女，硕士学位，医师。研究方向：脑血管疾病的中西医结合诊治。四川省中医药管理局科研专项（川中医药函〔2020〕196号）；西南医科大学附属中医医院联合项目（2018XYLH-006）；西南医科大学附属中医医院科研项目（西南医大中医院〔2019〕119号）；泸州市科技计划项目（2022-SYF-34）。
更新日期/Last Update: 2025-06-15