木蝴蝶茶活性成分提取工艺及对阿尔兹海默病小鼠认知能力干预作用

Extraction Technology of Active Components from Muhudie Tea and Intervention of Cognitive Ability on AD Mice

目的:优选木蝴蝶茶总黄酮的提取工艺,研究其对阿尔兹海默病(AD)小鼠认知能力的影响。方法:采用正交设计试验,以总黄酮收率为指标,以溶剂浓度、溶剂用量、提取次数、提取时间为主因素,优选最佳提取工艺;腹腔注射东莨菪碱(3 mg/kg)建造AD小鼠模型,木蝴蝶茶高、中、低剂量组分别给予1.928、3.857和5.785 g/kg,空白对照组、模型组给予等体积的生理盐水溶液,Morris水迷宫检测各组小鼠隐蔽平台实验潜伏期;末次给药后1 h,心脏取血测定血清生化指标lg G、SOD、MDA。结果:最佳提取工艺:药材加25倍量70%乙醇,回流提取2次,每次60 min。与空白对照组比较,各组小鼠隐蔽平台实验潜伏期差异显著(P<0.05),与模型组比较,高剂量组25 d后差异均显著(P<0.05),中剂量组第28 d差异显著(P<0.05)。与模型组比较,中剂量组可显著提高SOD活性(P<0.05),高剂量组lg G含量升高,SOD活性增强,MDA含量降低(均P<0.05)。结论:该工艺总黄酮得率较高,工艺稳定、重复性好,可为木蝴蝶茶的生产提供参考;木蝴蝶茶对AD小鼠空间学习记忆能力有一定的改善作用,其机制与血清中lg G、SOD和MDA相关。

Objective： To optimize extraction technology for total flavonoids of Muhudie Tea and investigate the effect of cognitive ability on Alzheimer＇s disease（ AD） mice. Methods： The optimum extraction condition was investigated by orthogonal design and regarded the yield of total flavonoids as index by using concentration of solvent,dosage of solvent,duration of extraction and frequency of extraction as the main factors. The AD model on mice was built by peritoneal injection of scopolamine（ 3 mg/kg）.Muhudie Tea was treated at the dose of 1. 928 g/kg,3. 857 g/kg and 5. 785 g/kg,respectively. The normal control and model groups were treated with equal volume of physiological saline. Morris water maze was used to detect the latency of hidden platform experiment on each group＇s mice. The last 1 h after the treatment,blood was taken out from the heart and serum biochemical indicators of lg G,SOD,MDA were measured. Results： The optimal extraction technique was to extract total flavonoids in 70% alcohol with 25 times weight of herbal medicine for 2 times,with 60 min once. Compared with the control group,the latency difference of the hidden platform was significant（ P〈0. 05）. Compared with the model group,the differences were significant in the high dose group after 25 d and the medium dose group in 28 d（ P〈0. 05）. Compared with the model group,Muhudie Tea could significantly increase the SOD activity at the dose of 3. 857 g/kg（ P〈0. 05） and make lg G content increased,SOD activity increased and MDA content decreased at the dose of 5. 785 g/kg（ P〈0. 05）. Conclusion： High yield of total flavonoids is obtained with the present technology. The results with better repeatability are stable,which can provide the reference for the production of Muhudie Tea. It has a certain improvement effect on the spatial learning and memory ability of AD mice and its mechanism is related to lg G,SOD and MDA in serum.