目的 探讨白头翁皂苷 B₄和常春藤皂苷 C（HSC，又名白头翁皂苷 B₅）质量比为 4∶1的组合物对帕金森病和阿尔茨海默病模型小鼠的保护作用及机制。方法 取60只健康雄性C57BL/6小鼠，随机抽取10只作为对照组，剩余50只小鼠ip1-甲基-4-苯基-1，2，3，6-四氢吡啶（MPTP）制备帕金森病（PD）模型，模型构建成功后随机分为模型组、左旋多巴胺片（阳性对照，30 mg·kg^-1）组和白头翁皂苷B₄＋HSC高、中、低剂量（20、10、5 mg·kg^-1）组，每组10只，ig给药14 d，每天1次，对照、模型组小鼠ig等量0.9%的氯化钠溶液。每2天观察动物震颤行为出现与消失的时间，于给药第7、14天展开行为学实验并记录评分；HE染色法观察脑组织黑质病理学变化；免疫组化检测脑组织 α-突触核蛋白表达；实时荧光定量 PCR（qRTPCR）检测小鼠脑组织多巴胺转运体（DAT）、酪氨酸羟化酶（TH）mRNA表达。将 50只 APP/PS1转基因小鼠随机分为模型组、盐酸多奈哌齐（阳性药，1.5 mg·kg^-1）组和白头翁皂苷B₄＋HSC高、中、低剂量（20、10、5 mg·kg^-1）组，取 10只同背景 APP/PS1转基因阴性小鼠作为对照组，每天1次，连续ig给药28 d，模型组和对照组ig等量0.9%的氯化钠溶液。给药结束后进行行为学实验并记录评分；ELISA法检测小鼠血清中炎症因子白细胞介素（IL-6、IL-1β）、肿瘤坏死因子-α（TNF-α）水平、环氧化酶 2（COX-2）和氧化应激因子活性氧（ROS），试剂盒检测超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）水平。结果 在 PD模型小鼠中，与模型组比较，白头翁皂苷 B₄＋HSC显著降低震颤麻痹评分（P＜0.01、0.001），显著降低旷场实验评分（P＜0.01、0.001），显著降低爬杆实验评分（P＜0.01、0.001），显著降低悬挂实验评分（P＜0.01、0.001），显著降低游泳实验评分（P＜0.01、0.001），显著降低行为学实验综合评分（P＜0.001），明显改善黑质体结构损伤，明显降低α-突触核蛋白表达，显著升高小鼠脑组织DAT和TH mRNA水平（P＜0.05、0.01、0.001）。在AD模型小鼠中，与模型组比较，白头翁皂苷 B₄＋HSC显著缩短水迷宫定位航行实验逃避潜伏期（P＜0.001），明显延长在目标象限的累计停留时间（P＜0.05、0.01、0.001），显著增加准确穿越平台所在位置的次数（P＜0.05、0.01），显著降低血清炎症因子水平（P＜0.05、0.01、0.001），显著降低 ROS水平、升高 SOD和 GSH-Px的水平（P＜0.05、0.01、0.001）。结论 白头翁皂苷＋HSC对 PD和 AD小鼠具有保护作用，可能通过发挥抗炎、抗氧化，改善脑组织和神经元损伤等来实现。

Objective To investigate the protective effects of a combination of Pulsatilla saponin B₄ and hederagenin C (HSC, anemoside B₅) with a quality ratio of 4∶ 1 on Parkinson's disease (PD) and Alzheimer's disease (AD) models in mice and the underlying mechanisms.Methods Sixty healthy male C57BL/6 mice were randomly selected, with 10 mice serving as the control group. The remaining 50 mice were intraperitoneally injected with 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) to establish a PD model. After the model was successfully established, the mice were randomly divided into the model group, the positive control group (levodopa hydrochloride tablets, 30 mg·kg^-1), and the Pulsatilla saponin B₄+HSC high, medium, and low dose groups (20, 10, and 5 mg·kg^-1), with 10 mice in each group. The mice were ig given the drugs for 14 d, once a day. The control and model groups were ig given an equal volume of 0.9% sodium chloride solution. The onset and disappearance times of tremor behavior were observed every two days. Behavioral experiments were conducted and scores were recorded on days 7 and 14 of the treatment. Hematoxylin and eosin (HE) staining was used to observe pathological changes in the substantia nigra of the brain tissue. Immunohistochemistry was used to detect the expression of α-synuclein in the brain tissue. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of dopamine transporter (DAT) and tyrosine hydroxylase (TH) mRNA in the brain tissue. 50 APP/PS1 transgenic mice were randomly divided into the model group, the positive drug group (donepezil hydrochloride, 1.5 mg·kg^-1), and the high, medium, and low dose groups of Pulsatilla saponin B₄+HSC (20, 10, and 5 mg·kg^-1), respectively. Ten APP/PS1 transgenic wild-type mice were taken as the control group. The mice were administered ig once a day for 28 d. The model group and the control group were given the same amount of 0.9% sodium chloride solution. After the administration, behavioral tests were conducted and the scores were recorded. The levels of inflammatory factors interleukin-6 (IL- 6), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), cyclooxygenase-2 (COX-2), and reactive oxygen species (ROS) in the serum of the mice were detected by ELISA. The levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were detected by kits.Results In the PD model mice, compared with the model group, Pulsatilla saponin B₄+HSC significantly reduced the tremor paralysis score (PD-RATS) score (P < 0.01, 0.001), significantly reduced the open field test score (P < 0.01, 0.001), significantly reduced the climbing rod test score (P < 0.01, 0.001), significantly reduced the hanging test score (P < 0.01, 0.001), significantly reduced the swimming test score (P < 0.01, 0.001), and significantly reduced the comprehensive score of behavioral test (P < 0.001), and significantly improved the structure damage of substantia nigra, significantly reduced the expression of α -synuclein, and significantly increased the levels of DAT and TH mRNA in the brain tissue of mice (P < 0.05, 0.01, 0.001). In the AD mouse model, compared with the model group, Pulsatilla saponin B₄ + HSC significantly shortened the escape latency in the water maze spatial navigation experiment (P < 0.001), significantly prolonged the cumulative stay time in the target quadrant (P < 0.05, 0.01, 0.001), significantly increased the number of accurate crossings of the platform location (P < 0.05, 0.01), significantly reduced the serum inflammatory factor level (P < 0.05, 0.01, 0.001), significantly reduced the level of ROS, and increased the level of SOD and GSH-Px (P < 0.05, 0.01, 0.001).Conclusion Pulsatilla saponin B₄+HSC effectively played protective roles on PD and AD in mice, which may be achieved through anti-inflammatory and antioxidant effects and improvement of brain tissue and neuron damage.

R285.5

广西高校引进海外高层次人才“百人计划”项目（05018064）