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(duì)帕金森病和阿爾茨海默病模型小鼠的保護(hù)作用及機(jī)制。方法 取60只健康雄性C57BL/6小鼠，隨機(jī)抽取10只作為對(duì)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(jī)分為模型組、左旋多巴胺片（陽(yáng)性對(duì)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(duì)照、模型組小鼠ig等量0.9%的氯化鈉溶液。每2天觀察動(dòng)物震顫行為出現(xiàn)與消失的時(shí)間，于給藥第7、14天展開(kāi)行為學(xué)實(shí)驗(yàn)并記錄評(píng)分；HE染色法觀察腦組織黑質(zhì)病理學(xué)變化；免疫組化檢測(cè)腦組織 α-突觸核蛋白表達(dá)；實(shí)時(shí)熒光定量 PCR（qRTPCR）檢測(cè)小鼠腦組織多巴胺轉(zhuǎn)運(yùn)體（DAT）、酪氨酸羥化酶（TH）mRNA表達(dá)。將 50只 APP/PS1轉(zhuǎn)基因小鼠隨機(jī)分為模型組、鹽酸多奈哌齊（陽(yáng)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(duì)照組，每天1次，連續(xù)ig給藥28 d，模型組和對(duì)照組ig等量0.9%的氯化鈉溶液。給藥結(jié)束后進(jìn)行行為學(xué)實(shí)驗(yàn)并記錄評(píng)分；ELISA法檢測(cè)小鼠血清中炎癥因子白細(xì)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(yīng)激因子活性氧（ROS），試劑盒檢測(cè)超氧化物歧化酶（SOD）和谷胱甘肽過(guò)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(píng)分（P＜0.01、0.001），顯著降低曠場(chǎng)實(shí)驗(yàn)評(píng)分（P＜0.01、0.001），顯著降低爬桿實(shí)驗(yàn)評(píng)分（P＜0.01、0.001），顯著降低懸掛實(shí)驗(yàn)評(píng)分（P＜0.01、0.001），顯著降低游泳實(shí)驗(yàn)評(píng)分（P＜0.01、0.001），顯著降低行為學(xué)實(shí)驗(yàn)綜合評(píng)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(dá)，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(shí)驗(yàn)逃避潛伏期（P＜0.001），明顯延長(zhǎng)在目標(biāo)象限的累計(jì)停留時(shí)間（P＜0.05、0.01、0.001），顯著增加準(zhǔn)確穿越平臺(tái)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(duì) PD和 AD小鼠具有保護(hù)作用，可能通過(guò)發(fā)揮抗炎、抗氧化，改善腦組織和神經(jīng)元損傷等來(lái)實(shí)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(jìn)海外高層次人才“百人計(jì)劃”項(xiàng)目（05018064）