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    丹龙醒脑方对血管性痴呆大鼠神经血管单元及突触可塑性的影响

    Effect of Danlong Xingnao Formula on Neurovascular Unit and Synaptic Plasticity in Vascular Dementia Rats

      摘要
    • 摘要:
      目的 探讨丹龙醒脑方对血管性痴呆(vascular dementia，VD)大鼠神经血管单元(neurovascular unit，NVU)及突触可塑性的影响。
      方法 SD大鼠采用改良2-VO法制备VD大鼠模型，随机分为模型组、尼莫地平组、丹龙醒脑方高剂量组(14.8 g·kg−1)、丹龙醒脑方中剂量组(7.4 g·kg−1)、丹龙醒脑方低剂量组(3.7 g·kg−1)，每组10只，分别给予相应药物灌胃。同时设置假手术组，等量生理盐水灌胃，共给药4周。Morris水迷宫试验检测各组大鼠认知功能；透射电镜观察海马组织NVU及突触超微结构；采用免疫组化法检测海马组织微血管密度(microvessel density，MVD)、生长相关蛋白43(growth associated protein 43，GAP43)、神经生长抑制因子A(neurite outgrowth inhibitor A，Nogo-A)、血管内皮生长因子(vascular endothelial growth factor，VEGF)、胶质纤维酸性蛋白(glial fibrillary acidic protein，GFAP)表达；Real-time PCR法检测海马组织突触素(synapsin，SYP)、突触后致密蛋白-95(postsynaptic dense protein-95，PSD-95) mRNA的表达；Western blotting检测海马组织SYP、PSD-95蛋白表达。
      结果 与假手术组比较，模型组大鼠学习、空间记忆能力明显下降(P<0.01)，海马组织NVU及突触超微结构出现明显的病理改变，Nogo-A、GFAP表达升高(P<0.01)，MVD、GAP43、VEGF表达降低(P<0.01)，SYP、PSD-95 mRNA及蛋白表达降低(P<0.01)。与模型组比较，丹龙醒脑方干预后可明显提高模型大鼠的学习、空间记忆能力，减轻NVU及突触超微结构的病理形态学损伤，降低Nogo-A、GFAP表达，增加MVD、GAP43、VEGF表达，上调SYP、PSD-95 mRNA及蛋白表达(P<0.05或P<0.01)。
      结论 丹龙醒脑方可能通过促进NVU和突触可塑性重塑改善VD大鼠学习记忆能力。

       

      Abstract:
      OBJECTIVE  To observe the effects of Danlong Xingnao Formula on neurovascular unit(NVU) and synaptic plasticity in rats with vascular dementia(VD).
      METHODS  SD rats were gave modified 2-VO method to establish VD rat model. SD rats were randomly divided into model group, nimodipine group, Danlong Xingnao Formula high-dose(14.8 g·kg−1), medium-dose(7.4 g·kg−1) and low-dose(3.7 g·kg−1) group with 10 rats in each group. Administer corresponding medication by gavage separately, at the same time, the sham operation group was set and given the same amount of normal saline, all rats were treated for 4 weeks. Morris water maze test was used to evaluate the spatial learning and memory abilities of rats in each group. Observation of NVU and synaptic ultrastructure in hippocampal tissue using transmission electron microscopy. Detection of microvessel density(MVD), growth associated protein 43(GAP43), neurite outgrowth inhibitor A (Nogo-A), vascular endothelial growth factor(VEGF), and glial fibrillary acidic protein(GFAP) expression in hippocampal tissue using immunohistochemistry. Real-time PCR method was used for detecting the expression of synapsin(SYP) and postsynaptic dense protein-95(PSD-95) mRNA in hippocampal tissue. Western blotting was used for detection of SYP and PSD-95 protein expression in hippocampal tissue.
      RESULTS  Compared with the sham operation group, the learning and spatial memory abilities of the model group were significantly decreased(P<0.01), significant pathological changes were observed in the NVU and synaptic ultrastructure of hippocampal tissue, Nogo-A and GFAP expression were increased(P<0.01), MVD, GAP43, VEGF expression were decreased(P<0.01), the expression of SYP, PSD-95 mRNA and protein in hippocampus were decreased(P<0.01). Compared with the model group, after the intervention of Danlong Xingnao Formula, the learning and memory ability of VD rats was significantly improved; the pathological morphological damage to NVU and synaptic ultrastructure was reduced, the expression of Nogo-A and GFAP was reduced, while the expression of MVD, GAP43, VEGF was increased, the mRNA and protein expression of SYP, PSD-95 in the hippocampus was up-regulated(P<0.05 or P<0.01).
      CONCLUSION  The Danlong Xingnao Formula may improve the learning and memory abilities of VD rats by promoting NVU and synaptic plasticity remodeling.

       

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