补肾还聪胶囊对血管性痴呆大鼠的影响

Pharmacological study the protective effects and its mechanism of Bushen Huancong capsule on vascular dementia in rats

目的:研究补肾还聪胶囊对血管性痴呆的防治作用,并初步探讨其可能的作用机理,为进一步开发成为防治血管性痴呆的中药新药打下基础。方法:SD大鼠永久性大脑中动脉闭塞导致的血管性痴呆,造模后24小时,将模型大鼠随机分为模型组、尼莫地平0.02 g/kg、补肾还聪胶囊20 g/kg、10 g/kg及5 g/kg组,同时设假手术组。每天灌胃给药1次,连续35天。利用Morris水迷宫对各组大鼠进行学习记忆能力检测,观测学习记忆能力,于行为学实验结束后第二天,取血测定超氧化物歧化酶(SOD),丙二醛(MDA),一氧化氮(NO),一氧化氮合酶(NOS),诱导性一氧化氮合酶(iNOS),取脑组织测定单胺氧化酶(MAO),乙酰胆碱(Ach),胆碱乙酰转移酶(ChAT),乙酰胆碱酯酶(AChE)的活性等指标。银染观察大鼠海马CA1区神经元染色。结果:Morris水迷宫实验显示,第一天各组大鼠逃避潜伏期无差异,其余几天模型组大鼠在定位航行试验中隐匿平台平均逃避潜伏期较正常组显著延长,补肾还聪胶囊20 g/kg,10 g/kg剂量组平均逃避潜伏期较模型组显著缩短,跨越原平台位置次数明显增加。空间探索试验中模型组跨越原平台位置次数,在原平台象限滞留时间及原平台象限内的游程占总游程的百分比明显减少。空间探索试验中,与模型组比较,补肾还聪胶囊20 g/kg剂量组平台象限滞留时间及平台象限游程占总游程的百分比显著增加。生化检测结果显示:模型组脑组织Ach,ChAT含量较假手术组显著降低,与模型组比较,补肾还聪胶囊20 g/kg,10 g/kg剂量组提高了Ach,ChAT含量。模型组大鼠血清SOD活性较假手术组明显降低,MDA、NO、NOS、iNOS活性较假手术组明显升高,补肾还聪胶囊20 g/kg及10 g/kg可显著对抗脑缺血所致上述指标的改变,与模型组比较有统计学差异。模型组大鼠脑组织MAO活性较假手术组明显升高,而这种升高可被补肾还聪胶囊20 g/kg,10 g/kg,5 g/kg剂量组抑制(P<0.05)。银染结果显示:补肾还聪胶囊明显改善血管性痴呆大鼠海马CAl区神经元数量的减少。结论:补肾还聪胶囊可显著改善VD大鼠的学习记忆功能,其作用机制可能与其具有抗氧化应激、改善胆碱能系统功能、减少海马区神经元的损伤有关。

To study the effects and mechanisms of Bushen Huancong Capsule on the vascular dementia in rats. Methods： Vascular de- mentia were induced by permanent middle cerebral artery occlusion（pMCAO） in male rats . 24 h later, the ischemic rats were divided into five groups. These rats were administrated by ig respectively Nimedipine 0.02 g/kg, Bushen Huancong Capsule 20 g/kg,10 g/kg, 5 g/kg, or the same volume of solvent（the model group and sham-operation group）. Once per day, for 35 d. The levels of learning and memory of rat were investigated by morris water maze. Meanwhile levels of malondialdehyde （ MDA ） and Nitric oxide（ NO）, activities of superexide dis- mutase （ SOD）, Nitric oxide synthesis（NOS） and Introduce nitric oxide synthase（iNOS） in serum, the content of Acetylcholine（Ach） and the activities of Choline acetyltransferase （CHAT） , Acetylcholine Esterase （AchE） and Monoamine oxidase（MAO） in brain issue were measured in rats. The CA1 neurocytes in the hippocampus were observed by Bielschowsky stain. Results： Morris Water Maze test： On the place navigation test, the latency of escape among groups have no difference at the first day,Bushen Huancong Capsule 20 g/kg and 10 g/kg can obviously decreased escape latencies time. In spatial probe test, the frequency of passing through the platform, the time spent in target quadrant and the percentage of the distance spent in target quadrant of the model group were obviously decreased as compared with the sham- operation group（ P 〈 0.05 or P 〈 0. 01 ） , Bushen Huancong Capsule 20 g/kg, 10 g/kg and 5 g/kg can increase the frequency of passing through the platform. And Bushen Huancong Capsule 20 g/kg also increase the time spent in the target quadrant and the percentage of the distance spent in the target quadrant in VD rats. Biochemical Test. The levels of Ach, ChAT were lower but the MAO was higher in the brain tissue from the model group than that in the sham-operation group in rats（ P 〈0.01 ）, and Bushen Huancong Capsule 20 g/kg ,10 g/kg sig- nificantly inceased levels of Ach, ChAT and decreased the activities of MAO in VD rats. The SOD activity were lower, but the levels of MDA, NO, NOS and iNOS were higher in serum from the model group than that in the sham-operation group（ P 〈 0.01 ）, Bushen Huancong Capsule 20 g/kg, 10 g/kg significantly improve theses indexes in VD rats（ P 〈0.05 or P 〈0.01 ）. Bielschowsky stain：The number of the neurocyte in hippocampal CAl area were obviously improved by Bushen Huancong Capsule in vascular dementia rats. Conclusion： Bushen Huancong Capsule could obviously improve the function of learning and memory of VD rats. The possible mechanisms may be due to Bushen Huancong Capsule has the actions of antioxidation, improvement of cholinergie system in brain, or lower the injury of neurons in the hippocampal CAl area.