醒脑启智胶囊对血管性痴呆小鼠治疗作用机理研究
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摘要
目的:血管性痴呆(以下简称VD)属中医学老年呆病范畴。近年来,随着社会人口老龄化,VD的患病率越来越高,成为严重影响人们生活质量的重要因素。因此,深入探讨VD的病理机制,开发和研制防治VD的有效药物具有非常重要的意义。根据中医学对VD的传统认识和现代研究成果,导师以补益肝肾,化痰祛瘀为法,研制成醒脑启智胶囊。采用双侧颈总动脉结扎的方法制备小鼠VD模型,观察了该方药对VD小鼠行为学、脑组织海马区神经递质、海马组织病理形态学、海马细胞增殖与凋亡及海马组织NGBmRNA等指标的影响,以进一步探讨其对VD的治疗作用机理。
方法:
第一部分醒脑启智胶囊对血管性痴呆小鼠行为学及脑组织病理形态学的影响
实验采用双侧颈总动脉结扎的方法制备血管性痴呆小鼠模型。健康雄性昆明小鼠360只,随机分为假手术组、模型组、醒脑启智胶囊高剂量组(以下简称“高剂量组”)、醒脑启智胶囊低剂量组(以下简称“低剂量组”)、银杏叶液对照组(以下简称“银杏叶组”)、尼莫地平液对照组(以下简称“尼莫地平组”)6组,每组60只。假手术组小鼠只分离颈总动脉,穿线但不结扎,尾部不放血,其余各组结扎双侧颈总动脉阻断血流加尾部放血,制备反复脑缺血再灌注VD模型。术后第2天开始治疗。高剂量组和低剂量组灌服醒脑液,银杏叶组灌服银杏叶液,尼莫地平组灌服尼莫地平液,假手术组和模型组均灌服生理盐水。分别于术后7、15、30d进行行为学实验(跳台法、水迷宫法)。行为学实验结束后断头处死小鼠,取左侧脑组织固定后切片,常规HE染色,光镜下观察脑组织的形态变化。
第二部分醒脑启智胶囊对血管性痴呆小鼠脑组织海马区神经递质的影响
Objective: Vascular Dementia (VD hereinafter) subordinates to the disease scope of dementia of old age in the light of TCM (tradditional Chinese medincine). In the recent years,with the aging of population the prevalence of vascular dementia is on the rise and affects seriously the people’s working ability as well as living quality. Therefore,it’s of great significance to make deeper research on it’s pathological mechanism and develop the effective medicine with the integration of Chinese and western medical theories. Based on the conventional knowledge and the modern research achievements of TCM on VD, we developed XingNaoQiZhi capsule with the function of benefitting liver and kidney, promoting blood circulation and resolving phlegm.In order to make probe into its treatment action mechanism, we adopted VD model of kunming mice with the operation of cerebral ischemic reperfusion , observed its effect on praxiology, neurotransmitter, pathomorphology, cell proliferation and Apoptosis of hippocampus, the expression of NGBmRNA, etc.
Methods:
Part One The effect of XingNaoQiZhi capsule on praxiology and cerebral pathomorphology of VD mice
We duplicated the model of VD mice with the cerebral ischemic-reperfusion operation with the ligation both side carotid artery. 360 Kunming mice were divided into six groups at random , which were sham-operation group,model group, high dose of XingNaoQiZhi capsule group(which were abbreviated to“High-dose group”in the following), low dose of XingNaoQiZhi capsule group(which were abbreviated to“low-dose group”in the following), extract of Ginkgobiloba control group(which were abbreviated to“Ginkgobiloba group”)and Nimodipine control group(which
方法:
第一部分醒脑启智胶囊对血管性痴呆小鼠行为学及脑组织病理形态学的影响
实验采用双侧颈总动脉结扎的方法制备血管性痴呆小鼠模型。健康雄性昆明小鼠360只,随机分为假手术组、模型组、醒脑启智胶囊高剂量组(以下简称“高剂量组”)、醒脑启智胶囊低剂量组(以下简称“低剂量组”)、银杏叶液对照组(以下简称“银杏叶组”)、尼莫地平液对照组(以下简称“尼莫地平组”)6组,每组60只。假手术组小鼠只分离颈总动脉,穿线但不结扎,尾部不放血,其余各组结扎双侧颈总动脉阻断血流加尾部放血,制备反复脑缺血再灌注VD模型。术后第2天开始治疗。高剂量组和低剂量组灌服醒脑液,银杏叶组灌服银杏叶液,尼莫地平组灌服尼莫地平液,假手术组和模型组均灌服生理盐水。分别于术后7、15、30d进行行为学实验(跳台法、水迷宫法)。行为学实验结束后断头处死小鼠,取左侧脑组织固定后切片,常规HE染色,光镜下观察脑组织的形态变化。
第二部分醒脑启智胶囊对血管性痴呆小鼠脑组织海马区神经递质的影响
Objective: Vascular Dementia (VD hereinafter) subordinates to the disease scope of dementia of old age in the light of TCM (tradditional Chinese medincine). In the recent years,with the aging of population the prevalence of vascular dementia is on the rise and affects seriously the people’s working ability as well as living quality. Therefore,it’s of great significance to make deeper research on it’s pathological mechanism and develop the effective medicine with the integration of Chinese and western medical theories. Based on the conventional knowledge and the modern research achievements of TCM on VD, we developed XingNaoQiZhi capsule with the function of benefitting liver and kidney, promoting blood circulation and resolving phlegm.In order to make probe into its treatment action mechanism, we adopted VD model of kunming mice with the operation of cerebral ischemic reperfusion , observed its effect on praxiology, neurotransmitter, pathomorphology, cell proliferation and Apoptosis of hippocampus, the expression of NGBmRNA, etc.
Methods:
Part One The effect of XingNaoQiZhi capsule on praxiology and cerebral pathomorphology of VD mice
We duplicated the model of VD mice with the cerebral ischemic-reperfusion operation with the ligation both side carotid artery. 360 Kunming mice were divided into six groups at random , which were sham-operation group,model group, high dose of XingNaoQiZhi capsule group(which were abbreviated to“High-dose group”in the following), low dose of XingNaoQiZhi capsule group(which were abbreviated to“low-dose group”in the following), extract of Ginkgobiloba control group(which were abbreviated to“Ginkgobiloba group”)and Nimodipine control group(which
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