目的 探究鎖陽(yáng)醋酸乙酯提取物（ECS）對(duì)阿爾茨海默癥（AD）模型小鼠行為學(xué)、線(xiàn)粒體組織形態(tài)學(xué)及線(xiàn)粒體動(dòng)力學(xué)相關(guān)蛋白表達(dá)的影響和作用機(jī)制。方法 采用8周齡AD模型5xFAD轉(zhuǎn)基因小鼠為模型組，8周齡野生型C57BL/6小鼠為對(duì)照組，ECS組為模型鼠ig ECS 47 mg/kg，每天1次，連續(xù)14 d。給藥6和12周分別進(jìn)行跳臺(tái)、水迷宮、曠場(chǎng)行為學(xué)檢測(cè)；給藥12周后取海馬部位進(jìn)行電鏡觀察；Western blotting檢測(cè)線(xiàn)粒體融合相關(guān)蛋白MFN1、OPA1和線(xiàn)粒體分裂相關(guān)蛋白DRP1的表達(dá)。結(jié)果 水迷宮結(jié)果顯示，給藥6周，3組間均無(wú)顯著性差異；給藥12周后，與對(duì)照組比較，模型組小鼠潛伏期時(shí)間延長(zhǎng)（P<0.05），穿越平臺(tái)次數(shù)顯著減少（P<0.05）；與模型組比較，ECS組小鼠潛伏期時(shí)間顯著縮短（P<0.05），穿越平臺(tái)次數(shù)顯著增加（P<0.05）；跳臺(tái)結(jié)果顯示，與對(duì)照組比較，給藥6周及12周，模型組小鼠跳臺(tái)潛伏期時(shí)間顯著縮短（P<0.05），跳臺(tái)次數(shù)顯著增多（P<0.05）；與模型組比較，給藥6周，ECS組無(wú)顯著性差異；給藥12周，ECS組跳臺(tái)潛伏期時(shí)間顯著延長(zhǎng)（P<0.05），跳臺(tái)次數(shù)顯著減少（P<0.05）。曠場(chǎng)結(jié)果顯示，給藥12周，三組均無(wú)顯著性差異。電鏡結(jié)果顯示，對(duì)照組線(xiàn)粒體結(jié)構(gòu)清晰膜完整，嵴排列整齊；模型組線(xiàn)粒體膜破裂、直徑變小、嵴扭曲模糊；ECS組線(xiàn)粒體嵴不完全變形，部分基質(zhì)出現(xiàn)空泡。Western blotting結(jié)果顯示，與對(duì)照組比較，模型組OPA1及MFN1蛋白表達(dá)水平顯著下調(diào)，DRP1蛋白表達(dá)水平顯著上調(diào)（P<0.05）；與模型組比較，ECS組MFN1、OPA1蛋白表達(dá)水平顯著上調(diào)（P<0.05、0.01），DRP1蛋白表達(dá)水平顯著下調(diào)（P<0.05）。結(jié)論 ECS顯著改善AD模型小鼠的行為學(xué)，機(jī)制可能與調(diào)節(jié)線(xiàn)粒體動(dòng)力學(xué)失衡相關(guān)。

Objective To investigate the effect and mechanism of the effective fraction of Cynomorium songaricum Ethyl Acetate Extract (ECS) on behavior, mitochondrial histomorphology and expression of mitochondrial kinetic-related proteins in Alzheimer's disease model mice. Methods 8-week-old AD model 5xFAD transgenic mice were used as the model group, 8-week-old wild-type C57BL/6 mice as the control group, and ECS as the administration group. After 6 weeks and 12 weeks of administration, the behaviors of platform jumping, water maze and so on were detected. After 12 weeks of administration, the hippocampus was taken and observed by electron microscope. Western blotting was used to detect the expression of MFN1, OPA1 and DRP1. Results The results of water maze showed that there was no significant difference among the three groups after 6 weeks of administration, but after 12 weeks of administration, the incubation period of the model group was longer than that of the control group (P < 0.05), and the number of crossing the platform was decreased (P < 0.05). Compared with the model group, the latency time and the number of crossing the platform in ECS group were shorter (P < 0.05) and higher than those in model group (P < 0.05). The results of platform jumping showed that compared with the control group, the latency time of platform jump in the model group was shortened (P < 0.05) and the number of platform jumping was increased (P < 0.05), which was similar to that in the model group. After 6 weeks of administration, there was no significant difference in ECS group, but at 12 weeks of administration, the latency time of platform jump in ECS group was prolonged and the number of platform jumping was decreased (P < 0.05). The results of open field showed that there was no significant difference among the three groups at 12 weeks. The results of electron microscope showed that the structure of mitochondria in the control group was clear and the crest was arranged neatly, while in the model group, the mitochondrial membrane ruptured, the diameter became smaller and the cristae were distorted and blurred. Western blotting results showed that compared with the control group, the expression level of OPA1 and mfn1 protein in the model group was significantly down regulated, and the expression level of drp1 protein was significantly up regulated (P < 0.05); compared with the model group, the expression level of mfn1 and OPA1 protein in the ECS group was significantly up regulated (P < 0.05, 0.01), and the expression level of drp1 protein was significantly down regulated (P < 0.05). Conclusion ECS can significantly improve the behavior of AD mice, and the mechanism may be related to the regulation of mitochondrial dynamics imbalance.

R965

北京市自然科學(xué)基金（面上項(xiàng)目）（18G40092）