人参糖蛋白对小鼠学习和记忆能力的影响

doi:10.13481/j.1671-587x.20160305

摘要/Abstract

摘要：

目的: 确定人参糖蛋白的基本化学组成,探讨其对东莨菪碱所致小鼠学习和记忆障碍的影响。方法: 采用中空纤维超滤设备,对已除去皂苷的人参水提取物进行分离。采用苯酚硫酸法测定总糖含量,Lowry法测定蛋白含量,高效液相色谱法(HPLC)测定人参糖蛋白的纯度和相对分子质量,将人参制备成1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生物后进行单糖组成分析。注射东莨菪碱建立小鼠记忆获得障碍模型,60只小鼠随机分为对照组(正常小鼠),模型组(训练后腹腔注射氢溴酸东莨菪碱2.5mg·kg^-1),阳性药组(训练前灌胃给药吡拉西坦9mg·kg^-1,30d),低、中、高剂量人参糖蛋白组(训练前灌胃给药,剂量为7.5、75.0、225.0mg·kg^-1),每组10只。水迷宫实验测定各组小鼠逃避潜伏期;跳台实验测定小鼠跨平台次数。结果: 人参糖蛋白总糖质量分数为58.41%,由7种单糖组成,其蛋白质量分数为19.93%,由17种氨基酸组成。人参糖蛋白相对分子质量分布在200~50000。Morris水迷宫实验,从第3天开始,与模型组比较,高剂量人参糖蛋白组小鼠平均逃避潜伏期明显缩短(P<0.05),跨平台次数明显增加(P<0.01)。在对位训练中,与模型组比较,高剂量人参糖蛋白组小鼠60s内在平台所在象限s的时间明显延长(P<0.01),从第2天起,与模型组比较,低、中和高剂量人参糖蛋白组小鼠平均逃避潜伏期明显缩短(P<0.05)。在跳台实验中,与模型组比较,低、中和高剂量人参糖蛋白组小鼠3min内的错误次数明显减少(P<0.05),中剂量人参糖蛋白组小鼠停留在平台上的潜伏期明显延长(P<0.05)。结论: 人参糖蛋白具有提高小鼠记忆能力的作用。

关键词: 人参, 糖蛋白, 学习能力, 记忆能力, 东莨菪碱

Abstract:

Objective: To identify the basic chemical compositions of panax ginseng glycoprotein,and to discuss its influence in the scopolamine-induced study and memory impairment in the mice. Methods: Hollow fiber ultrafiltration device was used to separate the ginseng aqueous extract which saponin had been removed.The content of total sugar was determined by phenol sulfuric acid method;the content of protein was determined by Lowry method;the purity and relative molecular weight distribution were determined by HPLC;the sugar compositions were analyzed after the preparation of PMP derivatives.The mouse model with acquired memory disorder was established through injection of scopolamine.A total of 60 mice were randomly divided into control group(normol mice),model group(intraperitoneal administration of scopolamine hydrobromide 2.5 mg·kg^-1 after training),positive drug group(intragastric administration of piracetam 9 mg·kg^-1 for 30 d before training),and low,middle and high doses of Panax ginseng glycoprotein groups (intragastric administration of glycoprotein 7.5,75.0,225.0 mg·kg^-1 respectively for 30 d before training),and there were 10 rats in each group.The escape latency and the number of crossing the platform of the mice were determined by Morris water maze and step-down test. Results: The total sugar mass fraction of Panax ginseng glycoprotein was 58.41% which was consisted of 7 kinds of sugars.The mass fraction of protein was 19.93% which was consisted of 17 kinds of amino acids.The relative molecular weights of Panax ginseng glycoprotein were 200-50 000.The Morris water maze test results showed that from the 3rd day,the average escape latency of mice in high dose of Panax ginseng glycoprotein group was significantly shortened compared with model group (P<0.05),and the number of crossing the platform in high dose of Panax ginseng glycoprotein group was significantly more than that in model group (P<0.01).The relative position test results showed that the time of spenting in the platform quadrant s of the mice within 60 s in high dose of Panax ginseng glycoprotein group was significantly prolonged compared with model group (P<0.05);from the 2 day,the average escape latency in different doses of Panax ginseng glycoprotein groups was significantly shortened compared with model group (P<0.05).The step-down test results showed that the mice in different doses of Panax ginseng glycoprotein groups made significantly less mistakes within 3 min compared with model group(P<0.05);compared with model group,the latent period of landing on the platform of the mice in middle dose of Panax ginseng glycoprotein group was significantly increased(P<0.05). Conclusion: Panax ginseng glycoprotein could improve the memory ability of mice.

Key words: Panax ginseng, glycoprotein, study ability, memory ability, scopolamine

中图分类号:

R285

罗浩铭, 陈英红, 周婷婷, 洪铁, 姜瑞芝, 王颖, 杨晓虹, 马莉. 人参糖蛋白对小鼠学习和记忆能力的影响[J]. 吉林大学学报(医学版), 2016, 42(03): 439-445.

LUO Haoming, CHEN Yinghong, ZHOU Tingting, HONG Tie, JIANG Ruizhi, WANG Ying, YANG Xiaohong, MA Li. Influence of glycoprotein from Panax ginsengin study and memory abilities in mice[J]. Journal of Jilin University Medicine Edition, 2016, 42(03): 439-445.

参考文献

[1] 赵莹,刘金平,卢丹,等.人参皂苷Re促进自然衰老大鼠学习记忆作用及其机理的研究[J]. 中药新药与临床药理,2007,18(1):20-22.

[2] 陈新梅,朱家壁.人参皂苷Rg1脂质体对东莨菪碱诱导大鼠学习记忆障碍的改善及其作用机制[J].中国临床药理学与治疗学,2005,10(8):898-902.

[3] 王晓英,陈霁,张均田.人参皂苷Rg1对β-淀粉样肽(25-35)侧脑室注射所致小鼠学习记忆障碍的改善作用及其机制[J].药学学报,2001,36(1):1-4.

[4] 张晶,王世荣,陈全成,等.人参皂苷Rg3(R),Rg3(S),Rg5/Rk1对乙醇致小鼠记忆阻碍改善作用的影响[J].吉林农业大学学报,2006,28(3):283-284.

[5] Sheng C,Peng W,Xia ZA,et al.The impact of ginsenosides on cognitive deficits in experimental animal studies of Alzheimer's disease:a systematic review [J].BMC Complement Alternat Med,2015,15:386.

[6] Liu J,Yan X,Li L,et al.Ginsenoside Rd improves learning and memory ability in APP transgenic mice [J].J Mol Neurosci,2015,57(4):522-528.

[7] 胡胜全,余惠旻,刘塔斯,等.人参水提物对β淀粉样蛋白诱导SH-SY5Y细胞凋亡的保护作用[J].时珍国医国药,2009,20(2):257-259.

[8] Hiroaki K,Hisao N,Teruko U,et al.Effects of nonsaponin fraction of red ginseng on learning deficits in aged rats [J].Physiol Behavior,2004,82(2/3):345-355.

[9] 王盛民,张瑛,孟建国,等.膨化加工对人参改善小鼠学习记忆能力的影响[J].陕西中医,2006,27(6):732-734.

[10] 张伟杰,王鹏,林茜,等.3种中药多糖的提取工艺[J].中国实验方剂学杂志,2011,17(16):19-23.

[11] Nelly BK,Gustar AH.New Method for quantitative determination of uronc acid [J].Anal Biochem,1973,54(2):484-487.

[12] 李建武,萧能赓,余瑞元,等.生物化学实验原理方法 [M].北京:北京大学出版社,1994.

[13] 王颖,高其品,李桂荣,等.人参糖肽PG-g的分离纯化与结构分析[J].天然产物研究与开发,2010,22(6):1053-1056.

[14] 杨兴斌,赵燕,周四元,等.柱前衍生化高效液相色谱法分析当归多糖的单糖组成[J].分析化学,2005,33(9):1287-1290.

[15] Wang H, Wei G,Liu F,et al.Characterization of Two Homogalacturonan Pectins with Immunomodulatory Activity from Green Tea [J].Int J Mol Sci,2014,15(6):9963-9978.

[16] 訾艳,王常青,党彦飞,等.白芸豆球蛋白的提取及其氨基酸组成研究[J].天然产物研究与开发,2014,26(8):1267-1271.

[17] Sloley BD,Pang PK,Huang BH,et al.American ginseng extract reduces scopolamine induced amnesia in a spatial learning task [J].JPsy Chiatry Neurosci,1999,24(5):442-452.

[18] Whishaw IQ.Formation of a place learning-set by the rat:A new paradigm for neurobehavioral studies [J].Physiol Behav,1985,35(1):139-143.

[19] 罗晓清,杨化新,杨洪淼,等.尖吻蝮蛇降纤酶寡糖链的甲基化分析[J].药物分析杂志,2010,30(3):357-360.

[20] 赵雅宁,刘文倩,牛静,等.参芎化瘀胶囊通过VEGF/Notch 1 信号通路改善大鼠缺血性脑卒中损伤[J].西安交通大学学报:医学版,2014,35(6):843-847.

[21] 王淑秀,常海敏,朱丰霞,等.人参皂苷Rg1对帕金森病小鼠黑质多巴胺能神经元凋亡及EphB1、TH、P-c-Jun蛋白表达的影响[J].郑州大学学报:医学版,2015,50(2):176-180.