石菖蒲-川芎配伍通过调节Nrf2/HO-1通路改善阿尔茨海默病的作用机制研究

Mechanisms of calamus chinensis-chuanxiong ameliorating Alzheimer’s disease by regulating the Nrf2/HO-1 pathway

关键词:  石菖蒲  川芎  阿尔茨海默病  氧化应激  Nrf2/HO-1信号通路

Key words:  calamus  chuanxiong  Alzheimer’s disease  oxidative stress  Nrf2/HO-1 signaling pathway

摘要:

[目的] 探讨石菖蒲-川芎水提液（SCP-CX）配伍对D-半乳糖联合亚硝酸钠（D-Galactose—Sodium nitrite，D-gal—NaNO₂）诱导的阿尔茨海默病（AD）模型小鼠的抗氧化应激和认知功能改善作用。[方法] 随机将8月龄C57BL/6小鼠分为Control组、Model组、给药组（盐酸多奈哌齐组（0.5 mg/kg）、SCP组（1.6 mg/kg）、CX组（0.8 mg/kg）、SCP-CX组（1.6 mg/kg与0.8 mg/kg），每组10只。通过皮下注射D-gal—NaNO₂建立AD模型小鼠，连续造模12周，造模第4周后开始灌胃给药。通过Morris水迷宫实验评价小鼠学习及空间记忆能力，通过试剂盒检测小鼠血清中谷胱甘肽过氧化物酶（GSH-PX）、丙二醛（MDA）含量和超氧化物歧化酶（SOD）活性，采用ELISA测定小鼠脑组织海马区活性氧（ROS）、β淀粉样蛋白_1-40（Aβ_1-40）含量，通过Western Blotting检测小鼠脑组织中核因子-红细胞2相关因子2（Nrf2）、血红素氧化酶1（HO-1）蛋白表达。[结果] 与Model组比较，各给药组逃避潜伏期显著缩短（P＜0.05），登上平台次数增多（P＜0.05）；GSH-Px水平显著上升（P＜0.05）；SOD活性显著下降（P＜0.05）；ROS含量显著降低（P＜0.05）；Aβ_1-40沉积显著减少（P＜0.05）。与Model组比较，SCP-CX组能显著增加Nrf2及HO-1蛋白表达量（P＜0.05）；与SCP组比较，SCP-CX组能显著增加Nrf2蛋白表达量（P＜0.05）。 [结论] SCP-CX能通过调节Nrf2/HO-1信号通路降低小鼠脑氧化应激反应，减少Aβ沉积，从而改善AD模型小鼠的认知功能障碍。

Abstract:

[Objective] To investigate the effects of water extract of acorus tatarinowii and ligusticum chuanxiong（SCP-CX） on anti-oxidative stress and cognitive function in mice with Alzheimer’s disease（AD） induced by D-galactose-sodium nitrite（D-gal—NaNO₂）. [Methods] Eight-month-old C57BL/6 mice were randomly divided into Control group，Model group，and drug administration group（Donepezil group（0.5 mg/kg），SCP group（1.6 mg/kg），CX group（0.8 mg/kg） and SCP-CX group（1.6 mg/kg vs. 0.8 mg/kg），each group with 10 mice. AD model mice were established by subcutaneous injection of D-gal—NaNO₂ for 12 consecutive weeks，and the drug was administered by gavage starting after the 4th week of modeling. The learning and spatial memory abilities of mice were evaluated by Morris water maze experiment，the serum levels of glutathione peroxidase（GSH-PX），malondialdehyde（MDA） and superoxide dismutase（SOD） activities of mice were detected by the kit，and the levels of reactive oxygen species（ROS） and β-amyloid_1-40（Aβ_1-40） in the hippocampal area of mouse brain tissue were determined by ELISA，and the protein expression of nuclear factor-erythroid 2 related factor 2（Nrf2），heme oxygenase 1（HO-1） in mouse brain tissue was detected by Western Blotting. [Results] Compared with the Model group，the escape latency was significantly shorter（P<0.05） and the number of ascending plateaus increased（P<0.05） in each dosing group；GSH-Px levels were significantly increased（P<0.05）；SOD activity was significantly decreased（P<0.05）；ROS levels were significantly reduced（P<0.05）；and Aβ_1-40 deposition was significantly reduced（P<0.05）. Compared with the Model group，the SCP-CX group could significantly increase Nrf2 and HO-1 protein expression（P<0.05）；compared with the SCP group，the SCP-CX group was able to significantly increase Nrf2 protein expression（P<0.05）. [Conclusion] SCP-CX can improve cognitive dysfunction in AD model mice by modulating the Nrf2/HO-1 signaling pathway to reduce oxidative stress and Aβ deposition in the mouse brain.