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目的:升降散对血管性痴呆(VD)大鼠学习记忆能力的影响及神经保护分子机制。方法:SD大鼠70只,随机分为正常组、假手术组,模型组,阳性药物组(盐酸多奈哌齐,1 mg·kg-1),升降散低、中、高剂量组(0.67,1.33,2.66 g·kg-1),每组10只,除正常组和假手术组外,其余各组采用改良的2VO法制作慢性脑缺血VD模型,手术1周后开始给药,连续灌胃给药8周。采用2VO两血管阻断法观察升降散对VD各组大鼠学习和记忆的影响、定位航行和空间探索行为功能,分析其学习记忆和空间辨认的能力;采用苏木素-伊红(HE)染色考察海马区病理形态;利用蛋白质免疫印迹(Western blot)和免疫组织荧光化学染色法检测血管内皮生长因子(VEGF),巢蛋白(Nestin),Wnt信号通路关键蛋白,磷酸化低密度脂蛋白受体相关蛋白(p-LRP),磷酸化糖原合成酶激酶(p-GSK3β)和β-连环蛋白(β-catenin)的表达水平,初步探讨升降散对VD大鼠海马组织保护作用机制。结果:与模型组比较,升降散不同剂量组能不同程度延长VD大鼠的避暗实验的潜伏期、缩短VD大鼠水迷宫定位航行和空间探索实验的潜伏期,升降散呈剂量依赖性促进海马区的病理性损伤修复及VEGF和Nestin表达,升降散呈剂量依赖性上调海马组织Wnt信号通路关键蛋白p-LRP,p-GSK3β和β-catenin蛋白水平,均具有一定的统计学意义(P<0.05,P<0.01)。结论:升降散改善VD大鼠学习记忆和空间辨认的行为学能力可能与激活海马组织Wnt信号通路活性有关。
Objective: The effect of Shengjiangsan on learning and memory of vascular dementia (VD) rats and the molecular mechanism of neuroprotection. Methods: Seventy SD rats were randomly divided into four groups: normal group, sham operation group, model group, positive drug group (donepezil hydrochloride, 1 mg · kg-1), ascending and descending low, middle and high dose groups (0.67, 1.33, 2.66 g · kg-1), 10 rats in each group. Except for the normal group and the sham operation group, all the other groups were made into chronic cerebral ischemia VD model by the modified 2VO method. Medicine for 8 weeks. The effects of Shengjiang San on learning and memory, positioning navigation and spatial exploration behavior of rats in VD group were observed by 2VO two-vessel occlusion method, and their learning and memory and spatial identification ability were analyzed. The hematoxylin and eosin (HE) staining The histopathology of the hippocampus was observed by Western blot and immunohistochemical staining. The expressions of VEGF, Nestin, Wnt signaling pathway and phosphorylated low density lipoprotein receptor (P-LRP), phosphorylated glycogen synthase kinase (p-GSK3β) and β-catenin (β-catenin) in rat hippocampus and to explore the protective mechanism of Shengjiang powder on hippocampus of VD rats. Results: Compared with the model group, the different doses of ascending and descending groups could prolong the latency of darkening test in VD rats, shorten the latency of navigation and space exploration experiment of water maze in VD rats, RSP could up-regulate the protein levels of p-LRP, p-GSK3β and β-catenin in Wnt signaling pathway in hippocampus in a dose-dependent manner (P <0.05) , P <0.01). Conclusion: Shengjiang San improve learning and memory and spatial cognition of VD rats behavioral abilities may be related to activation of Wnt signaling pathway activity in hippocampus.