目的 探討白頭翁皂苷 B₄和常春藤皂苷 C（HSC，又名白頭翁皂苷 B₅）質(zhì)量比為 4∶1的組合物對帕金森病和阿爾茨海默病模型小鼠的保護作用及機制。方法 取60只健康雄性C57BL/6小鼠，隨機抽取10只作為對照組，剩余50只小鼠ip1-甲基-4-苯基-1，2，3，6-四氫吡啶（MPTP）制備帕金森?。≒D）模型，模型構(gòu)建成功后隨機分為模型組、左旋多巴胺片（陽性對照，30 mg·kg^-1）組和白頭翁皂苷B₄＋HSC高、中、低劑量（20、10、5 mg·kg^-1）組，每組10只，ig給藥14 d，每天1次，對照、模型組小鼠ig等量0.9%的氯化鈉溶液。每2天觀察動物震顫行為出現(xiàn)與消失的時間，于給藥第7、14天展開行為學實驗并記錄評分；HE染色法觀察腦組織黑質(zhì)病理學變化；免疫組化檢測腦組織 α-突觸核蛋白表達；實時熒光定量 PCR（qRTPCR）檢測小鼠腦組織多巴胺轉(zhuǎn)運體（DAT）、酪氨酸羥化酶（TH）mRNA表達。將 50只 APP/PS1轉(zhuǎn)基因小鼠隨機分為模型組、鹽酸多奈哌齊（陽性藥，1.5 mg·kg^-1）組和白頭翁皂苷B₄＋HSC高、中、低劑量（20、10、5 mg·kg^-1）組，取 10只同背景 APP/PS1轉(zhuǎn)基因陰性小鼠作為對照組，每天1次，連續(xù)ig給藥28 d，模型組和對照組ig等量0.9%的氯化鈉溶液。給藥結(jié)束后進行行為學實驗并記錄評分；ELISA法檢測小鼠血清中炎癥因子白細胞介素（IL-6、IL-1β）、腫瘤壞死因子-α（TNF-α）水平、環(huán)氧化酶 2（COX-2）和氧化應激因子活性氧（ROS），試劑盒檢測超氧化物歧化酶（SOD）和谷胱甘肽過氧化物酶（GSH-Px）水平。結(jié)果 在 PD模型小鼠中，與模型組比較，白頭翁皂苷 B₄＋HSC顯著降低震顫麻痹評分（P＜0.01、0.001），顯著降低曠場實驗評分（P＜0.01、0.001），顯著降低爬桿實驗評分（P＜0.01、0.001），顯著降低懸掛實驗評分（P＜0.01、0.001），顯著降低游泳實驗評分（P＜0.01、0.001），顯著降低行為學實驗綜合評分（P＜0.001），明顯改善黑質(zhì)體結(jié)構(gòu)損傷，明顯降低α-突觸核蛋白表達，顯著升高小鼠腦組織DAT和TH mRNA水平（P＜0.05、0.01、0.001）。在AD模型小鼠中，與模型組比較，白頭翁皂苷 B₄＋HSC顯著縮短水迷宮定位航行實驗逃避潛伏期（P＜0.001），明顯延長在目標象限的累計停留時間（P＜0.05、0.01、0.001），顯著增加準確穿越平臺所在位置的次數(shù)（P＜0.05、0.01），顯著降低血清炎癥因子水平（P＜0.05、0.01、0.001），顯著降低 ROS水平、升高 SOD和 GSH-Px的水平（P＜0.05、0.01、0.001）。結(jié)論 白頭翁皂苷＋HSC對 PD和 AD小鼠具有保護作用，可能通過發(fā)揮抗炎、抗氧化，改善腦組織和神經(jīng)元損傷等來實現(xiàn)。

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

廣西高校引進海外高層次人才“百人計劃”項目（05018064）