中华中医药学刊

目的 探讨锁阳黄酮(Cynomorium songaricum flavonoids, CSF)对快速老化小鼠(SAMP8)认知功能障碍的影响及其作用机制。方法 选用50只7月龄雄性SAMP8小鼠，随机分为对照组，锁阳黄酮低、中、高剂量组[25、50、100 mg/(kg·d)]、多奈哌齐组。各剂量组给予锁阳黄酮对应剂量连续灌胃20 d,多奈哌齐组给予SAMP8小鼠多奈哌齐[0.71 mg/(kg·d)]灌胃20 d。给药结束后Morris水迷宫评价空间学习记忆表现，脑组织取材后进行尼氏染色观察海马锥体细胞形态并计数；Western blot测定各组海马区凋亡相关蛋白、醌氧化还原酶(NADPH:quinone oxidoreductase-1,NQO1)、核因子E2相关因子2(Nuclear factor E2 related factor 2,Nrf2)、Kelch环氧氯丙烷相关蛋白1(Kelch-like ECH-associated protein 1,Keap1)蛋白表达水平；ELISA测定血清谷胱甘肽过氧化物酶(Glutathione peroxidase, GSH-Px)和超氧化物歧化酶(Superoxide dismutase, SOD)活性、丙二醛(Malondialdehyd, MDA)的含量。结果 CSF显著增加了穿越平台次数与目标象限时间百分比，减小了上台前路程和逃避潜伏期(P<0.05);锁阳黄酮增加了海马CA1区正常锥体细胞数量，改善了锥体细胞的形态；Western blot结果表明，同对照组相比，锁阳黄酮中、高剂量组及多奈哌齐组海马区Nrf2、Keap1、NQO1蛋白表达量均显著升高(P<0.05);锁阳黄酮各剂量组、多奈哌齐组海马区Bcl-2/Bax蛋白表达比值均明显增加(P<0.05),锁阳黄酮中、高剂量组及多奈哌齐组海马区Caspase-3蛋白表达量均显著降低(P<0.05)。与对照组相比，锁阳黄酮提高了SAMP8小鼠血清SOD、GSH-Px的活性(P<0.05),降低了MDA的含量(P<0.05)。结论 锁阳黄酮能显著改善阿尔茨海默病(Alzheimer disease, AD)空间学习记忆障碍，其机制可能与抑制氧化应激与细胞凋亡相关。

Objective To investigate the effect and mechanism of Cynomorium songaricum flavonoids(CSF) on cognitive dysfunction in senescence-accelerated mice prone 8(SAMP8). Methods Fifty 7-month-old male SAMP8 were randomly divided into control group, low, intermediate and high dose groups of CSF[25,50,100 mg/(kg·d)],and donepezil group. The low, intermediate and high dose groups were given a corresponding dose of CSF to SAMP8 mice by continuous gavage for 20 days, while the donepezil group was given donepezil[0.71 mg/(kg·d)] to SAMP8 mice by gavage for 20 days. The spatial learning and memory performance was evaluate by the Morris water maze experiment after administration, and the morphology and count of hippocampal pyramidal cells was observed by Nissl staining. The apoptosis-related protein expression levels and the levels of nuclear factor E2 related factor 2(Nrf2),Kelch epichlorohydrin associated protein 1(Keap1) and quinone oxidoreductase-1(NQO1) in the hippocampus of each group were detected by Western blotting. The serum levels of malondialdehyde(MDA) content as well as glutathione peroxidase(GSH-Px) and superoxide dismutase(SOD) activities were measured by ELISA. Results CSF significantly reduced the escape latency and path length, while increased the number of crossing platforms and the percentage of target quadrant time(P<0.05). CSF improved the morphology and quantity of pyramidal cells in hippocampal CA1 region. Western blot results showed that compared with the control group, the protein expression levels of Nrf2,Keap1 and NQO1 in the hippocampus of the intermediate dose group, high dose group and donepezil group all significantly increased(P<0.05),the ratio of Bcl-2/Bax protein expression in the hippocampus of all administration groups of CSF and donepezil group increased substantially(P<0.05),and the expressions of Caspase-3 protein in the hippocampus of the intermediate dose group, high dose group and donepezil group all decreased significantly(P<0.05). Compared with the control group, CSF significantly increased the serum activities of SOD and GSH-Px in SAMP8 mice(P<0.05),while remarkedly decreased the content of MDA(P<0.05). Conclusion CSF can significantly improve spatial learning and memory impairment in Alzheimer's disease, and its mechanism may be related to inhibiting oxidative stress and cell apoptosis.