龙眼果肉提取物改善东莨菪碱诱导小鼠学习记忆功能

doi:10.3864/j.issn.0578-1752.2016.21.014

摘要/Abstract

摘要： 【目的】比较不同剂量龙眼果肉醇提物和水提物对东莨菪碱所致记忆获得性障碍小鼠学习记忆功能的影响，初步解释龙眼果肉改善学习记忆的作用机制。【方法】分析龙眼果肉醇提物和水提物中主要活性物质含量。将无特定病原体（SPF级）雄性昆明小鼠随机分为空白组、模型组、龙眼果肉醇提物低剂量组/高剂量组（150 mg·kg^-1/300 mg·kg^-1）、龙眼果肉水提物低剂量组/高剂量组（150 mg·kg-1/300 mg·kg^-1），共6组，连续灌胃28 d后，空白组腹腔注射等量生理盐水，各试验组腹腔注射东莨菪碱造模，0.5 h后，以潜伏期和穿梭次数为考察指标，采用避暗试验进行行为学测试，作为习得成绩；24 h后再次进行避暗试验，对小鼠进行记忆保持测试，测试结束后摘眼球取血，脱颈致死，取脑组织进行生化指标检测。测定脑组织的胆碱类（乙酰胆碱转移酶choline acetyltransferase，ChAT和乙酰胆碱酯酶acetylcholinesterase，AChE活力）及抗氧化相关指标（超氧化物歧化酶（superoxide dismutase，SOD）、谷胱甘肽过氧物酶（glutathione peroxidase，GSH-Px）活力及丙二醛（malondialdehyde，MDA）含量），血清中抗氧化相关指标（SOD、GSH-Px活力及MDA含量）。通过综合比较各组间行为学试验指标、脑组织、血清生化指标，判断龙眼果肉改善学习记忆功能的作用效果及作用机制。【结果】分析龙眼果肉提取物成分发现，龙眼果肉水提物主要为多糖及蛋白质，醇提物除含有糖类外，还含有较丰富的酚类、黄酮类和磷脂物质，其中，醇提物中总酚、总黄酮、总磷脂含量显著高于水提物的（P＜0.05）。行为学试验表明，模型组小鼠在避暗试验中5 min内穿梭次数为2.89次，是正常小鼠的6.09倍，高剂量龙眼果肉醇提物和水提物组小鼠穿梭次数分别为0.75次和0.56次，与正常组无显著差异；痴呆小鼠在避暗试验中的潜伏期仅为正常组的0.43倍,与模型组相比，各剂量龙眼果肉醇提物和水提物均能显著增加痴呆小鼠在避暗试验中的潜伏期（P＜0.05），且存在一定剂量效应关系，其中龙眼果肉醇提物和水提物高剂量组小鼠的避暗潜伏期分别为289.18 s、290.80 s，与正常组相比无显著差异（P＞0.05）。生化指标方面，与模型组相比，各剂量龙眼果肉醇提物和水提物均能显著增加痴呆小鼠脑组织中ChAT活力，显著降低AChE活力（P＜0.05），且存在一定的剂量效应关系。相同剂量时，龙眼果肉水提物效果优于醇提物，其中龙眼果肉醇提物高剂量组，龙眼果肉水提物低、高剂量组小鼠脑组织中ChAT活力基本恢复正常水平；在抗氧化水平方面，与模型组相比，龙眼果肉醇提物高剂量组显著增加痴呆小鼠脑组织和血清中SOD、GSH-Px活力，显著降低MDA含量（P＜0.05），基本达到正常水平，效果优于同等剂量的龙眼果肉水提物。【结论】龙眼果肉水提物和醇提物均可改善东莨菪碱诱导记忆获得性障碍小鼠的学习记忆功能，但二者的作用机制可能存在差异，龙眼果肉水提物主要通过调节胆碱能神经系统改善学习记忆功能，其主要作用的物质基础可能为多糖或糖蛋白；醇提物则主要通过提高机体抗氧化活力改善学习记忆功能，其主要作用物质可能为多酚、黄酮、磷脂等。

关键词: 龙眼果肉；东莨菪碱；学习记忆功能；乙酰胆碱转移酶, 抗氧化

Abstract: 【Objective】The objective of this experiment is to study the effect of different doses of both ethanol extract and aqueous extract of longan pulp on learning and memory function in dementia mice induced by scopolamine and to clarify the mechanism of longan pulp in improving learning and memory of imparment mice.【Method】The contents of the main active substances in the ethanol extract and water extract of longan pulp were analyzed. SPF male Kunming mice were randomly divided into 6 groups for 28 d administration: normal control group, model group, low/high dose longan pulp ethanol extract group (150 mg?kg^-1/300 mg?kg^-1), low/high dose longan pulp aqueous extract group (150 mg?kg^-1/300 mg?kg^-1). At the end of last time administration, the passive avoidance tests were performed after mice received scopolamine intraperitoneal injection, the indexes of step-through latency and shuttle number were marked as acquisition trial, the passive avoidance tests were performed again after 24 h which was marked as retention trial. Then detection of the antioxidant activities of serum and brain, such as SOD, GSH-Px, MDA, as well as ChAT and AChE of brain tissue were carried out. 【Result】By analyzing the constituents of longan pulp extract, it was found that longan pulp aqueous extract contains mainly polysaccharide and protein, and ethanol extract contains not only carbohydrates, but also abundant phenolic acid, flavonoid and phospholipid substances. Moreover, the content of total phenols, total flavonoids and total phospholipid in ethanol extraction was significantly higher than that of aqueous extract. The shuttle number of model mice in acquisition trial of passive avoidance test in 5 min (2.89) was 6.09 times of normal control (0.46). Meanwhile shuttle numbers of the high dose longan pulp ethanol extract and aqueous extract groups were 0.75 and 0.56, respectively, the numbers had no significant difference with normal control. The step-through latency of model group in acquisition trial of passive avoidance test was decreased to 130 s (normal control group was 300 s, P＜0.05). Compared with model group, each dose of longan pulp ethanol extract and aqueous extract could significantly increase the dementia mice in the step-through latency of passive avoidance test (P＜0.05) with dose-dependent, especially the high dose longan pulp ethanol extract and aqueous extract group (289.18 s, 290.80 s, respectively,) basically reached normal level. Biochemical examinations revealed that each dose of longan pulp ethanol extract and aqueous extract could significantly increase the activity of ChAT in brain of dementia mice and significantly reduce AChE activity (P＜0.05) in a dose-dependent manner. At the same dose level, aqueous extract of longan pulp performed better than ethanol extract. ChAT activity recovered normal in high dose longan pulp ethanol extract group, low/high dose longan pulp aqueous extract groups. In the antioxidation, compared with the model group, longan pulp ethanol extract in high dose group was significantly increased SOD and GSH-Px activities and decreased MDA content significantly in serum and brain of dementia mice (P＜0.05), basically reached normal, the effect was better than the same dose of aqueous extract of longan pulp.【Conclusion】Both ethanol extract and aqueous extract of longan pulp have repairing effects on memory deficits and behavioral disorders produced by scopolamine. It was suggested that ethanol extract and aqueous extract of longan pulp may be bene?cial to the treatment of Alzheimer’s disease, but their action mechanisms might be different: to promote learning and memory, aqueous extract of longan pulp mainly regulates cholinergic nerve systemby polysaccharide or glycoprotein, and ethanol extract is mainly by means of providing antioxidation protection, phenolic acid, flavonoid and phospholipid are principal bioactive substances.

Key words: longan pulp, scopolamine, learning and memory function, choline acetyltransterase, antioxidation

白亚娟，刘磊，张瑞芬，邓媛元，黄菲，张名位. 龙眼果肉提取物改善东莨菪碱诱导小鼠学习记忆功能[J]. 中国农业科学, 2016, 49(21): 4203-4213.

BAI Ya-juan, LIU Lei, ZHANG Rui-fen, DENG Yuan-yuan, HUANG Fei, ZHANG Ming-wei. Longan Pulp Extracts Ameliorate Scopolamine-Induced Learning and Memory of Impairment Mice[J]. Scientia Agricultura Sinica, 2016, 49(21): 4203-4213.

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