沙棘果多酚提取工艺优化及其抗阿尔茨海默症活性

目的:确定沙棘果多酚(SBP)最佳提取工艺,并探究其醇提取物抗阿尔茨海默症(AD)活性。方法:以单因素实验为基础,星点设计-效应面法(CCD-RSM)优化沙棘果乙醇提取物提取工艺。60只小鼠随机分为空白组、阳性组、模型组及SBP高、中、低剂量组,D-半乳糖联合三氯化铝诱导AD模型。利用Morris水迷宫进行神经行为学检测,酶联免疫吸附测定小鼠脑中乙酰胆碱(Ach)、胆碱乙酰转移酶(ChAT)及乙酰胆碱酯酶(AchE)含量,HE染色观察脑海马区形态变化。结果:SBP最佳提取工艺为乙醇浓度为71%,超声时间32 min,液料比20:1 mL/g,SBP提取量是3.0813 mg/g。与模型组比较,SBP高剂量组AD小鼠逃避潜伏期缩短(P<0.01),探索潜伏期显著延长,探索次数显著减少(P<0.05),Ach含量极显著增加(P<0.01),AchE含量极显著减少(P<0.01),ChAT含量显著增加(P<0.05),小鼠脑海马区形态明显改善。结论:SBP提取工艺优化,实际值与预测值吻合度高,可行性强;SBP能够调节改善学习记忆能力,具有良好的抗AD活性。

Optimization of Extracting Process and Anti-alzheimer's Disease Activity of Polyphenols from the Fruit of Seabuckthorn(Hippophae rhamnoides L.)

Objective:To determine the optimum extraction conditions of polyphenols from sea buckthorn(Hippophae rhamnoides L.)fruit(SBP)and preliminarily evaluate its anti-alzheimer's disease activity. Methods:On the basis of single factor experiments,the optimum extraction conditions of SBP were investigated by central composite design-response surface(CCD-RSM). 60 mice were randomly divided into control group,positive group model group and high-dose,medium-dose,low-dose group of SBP. AD model was induced by D-galactose combined with aluminum chloride. The escape incubation period of mice were recorded learning. Exploration incubation period and the times of exploration were recorded after last administration. ELISA was used to determine the content of acetylcholine(Ach),acetylcholine esterase(AChE)and choline acetyltransferase(ChAT)in mice brain and observation of hippocampal neuronal damage in mice brain by hematoxylin-eosin staining. Results:The optimal extraction conditions were as follows:71% ethanol as extraction solvent,a liquid/solid radio of 20:1 mL/g and an extraction time of 32 min. Under these optimized conditions,the extraction yield of polyphenols was 3.0813 mg/g. Compared with the model group,the escape incubation period was shortened extremly significant(P<0.01),exploration latency was prolonged and the number of explorations was decreased in Morris water maze experiment significantly(P<0.05),the morphological changes in hippocampal region was improved and significantly increased content of Ach content extremly significant(P<0.01),extremly significant decreased content of AchE content(P<0.01),and significantly increased content of ChAT content(P<0.05)in the high-dose group of SBP. Conclusion:Using central composite design-response surface to optimize SBP alcohol extraction process,the actual and predicted values are consistent with high and good predictability which is feasible. The SBP have good anti-Alzheimer's disease activity.