丹参7种水溶性有效成分配伍对脑缺血再灌注所致记忆损伤模型小鼠的影响

Inf luence of Radix Salviae Miltiorrhizae Hydrosoluble Combinations on the Memory Dysfunction Induced by Ischemia-Reperfusion in Mice

目的:探讨前期研究筛选出的丹参七种水溶性有效成分最优配伍组合样品A2、B4、C11对脑缺血再灌注(ischemia reperfusion)小鼠学习记忆的保护作用。方法:采用改进的Himori法暂时性阻断两侧颈总动脉制备小鼠脑缺血再灌注损伤模型,应用水迷宫实验,观察配伍组合样品对脑缺血再灌注小鼠记忆功能的保护作用,检测小鼠断头耐缺氧存活时间,脑组织中超氧化物歧化酶(superoxide dismutase,SOD)、过氧化氢酶(catalase,CAT)、乙酰胆碱酯酶(acetylcholine esterase,ACh E)活力及丙二醛(malondialdehyde,MDA)含量。免疫组织化学方法检测小鼠海马神经元凋亡相关天冬氨酸特异性半胱氨酸蛋白酶-3(cysteine-asparate protease-3,caspase-3)和神经生长因子(nerve growth factor,NGF)的表达。结果:丹参7种水溶性有效成分配伍组合样品B4、C11可明显改善小鼠脑缺血再灌注所致的记忆障碍,增强耐缺氧能力,提高脑内SOD,CAT活性,降低ACh E活性和MDA含量;抑制凋亡蛋白caspase-3表达,上调NGF表达。其中以B4(2.8μmol·L-1)、C11(30.0μmol·L-1)组作用最为明显(P<0.01),A2各个剂量组效果不显著(P>0.05)。结论:丹参7种水溶性有效成分配伍组合样品B4、C11对脑缺血再灌注损伤小鼠记忆功能有保护作用,其机制可能通过提高清除脑内自由基能力,抑制小鼠神经细胞凋亡,促进神经再生,来减轻脑缺血再灌注引起的脑组织损伤。

Objective： To study the protective effects of Radix Salviae Miltiorrhizae Hydrosoluble combinations on cerebral ischemia and reperfusion injury and its mechanism. Methods： Cerebral ischemia and reperfusion model was conducted referring to improved Himori method in which bilateral common carotid arteries were temporarily obstructed. The effect of combination on learning and memory was examined in Morris water maze test. Anoxia survival time of decapitated mice, the activities of SOD, CAT and ACHE, and the content of MDA were measured. The influences of combination were studied on the neurological function and the expression of apoptosis related protein caspase-3 in hippocampal neuron and nerve growth factor in cortex using immunohistochemistry. Results： Combinations （B2. C11） could obviously improve memory impairment induced by cerebral ischemia and reperfusion in mice, enhance hypoxia tolerance, increase the activities of SOD and CAT, with decreasing the activities of AChE and the content of MDA. At the same time, the expression of caspase-3 was inhibited, increasing NGF. Effects of combination were most obvious in group （B4 1.4 μmol-L-1, Cll 30.0 μmol. L-1）, A2 combination has no significant effect at any dose. Conclusion： Combinations （B4、C11） have a significantly protective effect on hippocampal neurons after cerebral ischemia and reperfusion injury. The mechanism may be related to removing free radicals, inhibiting apoptosis of nerve cells, and promoting nerve regeneration.