定志小丸有效成分对阿尔兹海默病转基因模型小鼠的氧化应激、炎症和自噬因子的作用研究

Effects of the active ingredients of Dingzhi Xiaowan on oxidative stress,inflammation and autophagy factors in transgenic mice with Alzheimer's disease

目的研究定志小丸有效成分对阿尔兹海默病转基因模型小鼠的氧化应激、炎症和自噬因子的作用。方法将70只APP/PS1转基因小鼠随机分为7组,每组10只,即模型组、人参皂苷Rb3组(15 mg/kg),β-细辛醚组(45 mg/kg),联合组(人参皂苷Rb315 mg/kg加β-细辛醚45 mg/kg),多奈哌齐组(3 mg/kg)、Akt抑制剂组(25 mg/kg)和自噬抑制剂组(30 mg/kg),并设正常组(C57BL/6小鼠),每组10只。除模型组和正常组给予等体积生理盐水,给药30 d,每天1次。给药结束后,ELISA检测各组皮质中单胺氧化酶(MAO)、活性氧(ROS)、8-羟基脱氧鸟苷(8-OHDG)、肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)、白介素-10(IL-10)和Beclin-1的含量;苏木素伊红染色观察皮质的组织病理学情况。结果与正常组比较,模型组的MAO、ROS、8-OHDG、TNF-α、IL-6和Beclin-1含量增加,IL-10含量减少(P<0.01)。与模型组比较,人参皂苷Rb3组、β-细辛醚组、联合组和自噬抑制剂组MAO、ROS、8-OHDG、TNF-α、IL-6和Beclin-1含量减少,IL-10含量增加(P<0.01)。还有模型组和Akt抑制剂组的细胞间隙较大,细胞核核固缩明显,细胞排列较为稀疏和杂乱,人参皂苷Rb3组、β-细辛醚组、联合组、多奈哌齐组和自噬抑制剂组的细胞状态都有所缓解。结论定志小丸有效成分对APP/PS1转基因小鼠的具有抗氧化和抗炎的作用,其机制可能是通过激活Akt抑制自噬从而保护皮质神经元细胞。

Objective To investigate the effects of the active ingredients of Dingzhi Xiaowan on oxidative stress, inflammation and autophagy factors in transgenic mice with Alzheimer’s disease. Methods A total of seventy APP/PS1 transgenic mice were randomly divided into 7 groups, namely a model group, a ginsenoside Rb3 group(15 mg/kg), a β-asarone group(45 mg/kg), a combination group(ginsenoside Rb3 15 mg/kg plus β-asarone 45 mg/kg), a donepezil group(3 mg/kg), an Akt inhibitor group(25 mg/kg), an autophagy inhibitor group(30 mg/kg), and a normal group(C57BL/6 mice), with 10 mice in each group. Except the model group and the normal group, all the mice were given the equal volume of normal saline for 30 days, once a day. After the intervention, the contents of monoamine oxidase(MAO), reactive oxygen species(ROS), 8-hydroxydeoxyguanosine(8-OHDG), tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), interleukin-10(IL-10) and Beclin-1 in the cortex of each group were detected by ELISA, and the histopathology of the cortexes was observed by hematoxylin eosin staining. Results Compared with the normal group, the contents of MAO, ROS, 8-OHDG, TNF-α, IL-6 and Beclin-1 in the model group were increased, while the content of IL-10 was decreased(P<0.01). Compared with the model group, the contents of MAO, ROS, 8-OHDG, TNF-α, IL-6 and Beclin-1 in the ginsenoside Rb3 group, the β-asarone group, the combination group and the autophagy inhibitor group were decreased(P<0.01). In addition, the model group and the Akt inhibitor group had a larger cell space, an obvious nuclear pyknosis, a sparse and disordered cell arrangement. The cell states were alleviated in the ginsenoside Rb3 group, the β-asarone group, the combination group, the donepezil group and the autophagy inhibitor group. Conclusion The active ingredients of Dingzhi Xiaowan have antioxidant and anti-inflammatory effects on APP/PS1 transgenic mice, whose mechanism may be to protect cortical neurons by activating Akt and inhibiting autophagy.