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桂枝汤挥发油的成分归属分析及其对认知障碍小鼠学习记忆的影响

Analysis and adscription of volatiles fromGuizhi Tang using gas chromatography-mass spectroscopy and improvement of the learning and memory in mice

摘要目的 探讨桂枝汤挥发油成分归属及其对认知障碍模型小鼠学习记忆的影响.方法 采用水蒸气蒸馏法分别提取生姜、桂枝及桂枝汤挥发油,采用气相色谱-质谱联用法分析挥发油成分.小鼠按体重分层随机分为正常组、模型组、多奈哌齐组、桂枝汤挥发油低剂量组、桂枝汤挥发油高剂量组.多奈哌齐组灌胃2 mg/kg多奈哌齐溶液,桂枝汤挥发油低、高剂量组灌胃桂枝汤挥发油溶液5、20 mg/kg,药品均以0.5%的羧甲基纤维素钠悬浮至所需浓度,正常组与模型组灌胃等体积0.5%的羧甲基纤维素钠溶液,1次/d,连续给药15 d.除正常组外,其余各组小鼠于给药第8天腹腔注射东莨菪碱1 mg/kg制备小鼠记忆障碍模型.采用Morris水迷宫、跳台法检测学习记忆行为学,ELISA法测定小鼠血清中胆碱酯酶(cholinesterase,chE)、乙酰胆碱酯酶(acetyl cholinesterase,AchE)、丁酰胆碱酯酶(butyryl cholinesterase,BchE)、乙酰胆碱(acetylcholine,Ach)含量.结果 从桂枝汤挥发油中共鉴定38个化合物,来源于桂枝的9个成分加权占比约44%,来源于生姜的13个成分约占全方挥发油的18%.与模型组比较,桂枝汤挥发油低剂量组在目标象限游泳距离比例[(26.74±16.42)%比(9.42±8.50)%]、目标象限滞留时间的比例[(43.51±25.12)%比(14.50±12.23)%]增加(P<0.05),穿台次数[(1.63±1.19)比(0.25±0.46)]明显增加(P<0.05),跳台试验中犯错误次数[(0.57±0.98)比(4.43±2.40)]降低(P<0.05);桂枝汤挥发油低、高剂量组小鼠血清chE[(140.90±3.27)U/ml、(144.79±6.71)U/ml比(134.49±3.36)U/ml]、AchE[(3.30±1.31)U/ml、(3.94±0.78)U/ml比(8.52±3.39)U/ml]、BchE[(3.22±0.45)U/ml、(3.66±0.53)U/ml比(7.99±0.79)U/ml]水平降低(P<0.05),Ach[(4.10±0.38)ng/ml、(3.03±0.25)ng/ml比(1.72±0.50)ng/ml]水平升高(P<0.05).结论 桂枝汤挥发油主要来源于桂枝和生姜,该部位可通过拟胆碱效应改善东莨菪碱所致小鼠学习认知障碍.

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abstractsObjective Analysis and adscription of volatiles from Guizhi Tang and study on its improvement of the learning and memory in dementia mice induced by scopolamine.Methods The volatile oil from Guizhi Tang(GZT),Guizhi and Shengjiang was extracted using steam distillation method and was analyzed by GC-MS. Morris water maze and step-down test were carried out for obtain the difference of the learning and memory improvement in 40 ICR mice from randomized groups, such as the control group, the model group, the donepezil group (2 mg/kg), the low dose of volatile oil of GZT (5 mg/kg), and the high dose of volatile oil of GZT (20 mg/kg), and ACh, AchE, BchE and chE in serum were detected by ELISA. Results Among 38 identyfied volatile ingredients from GZT, 18(44% in weight) was from Guizhi, and 9 was from Shengjiang. Compared with the model group, the low and high dose of GZT volatile oil significantly increased swimming distance ratio in destination quadrant (26.74% ± 16.42%vs.9.42% ± 8.50%, P<0.05); goal quadrant time scale (43.51% ± 25.12%vs. 14.50% ± 12.23%,P<0.05)) increased significantly than the model group ; the number of errors in the experiment platform (1.63 ± 1.19vs. 0.25 ± 0.46, P<0.05) obviously increased than model group ; platform test in the made errors times (0.57 ± 0.98vs. 4.43 ± 2.4, P<0.05) significantly reduced. The GZT total volatile oil groups significantly reduced cognitive obstacles small rat serum in the cholinester enzyme (chE) (140.90 ± 3.27, 144.79 ± 6.71vs. 134.49 ± 3.36,P<0.05); acetylcholinesterase (AchE) (3.30 ± 1.31, 3.94 ± 0.78 vs.8.52 ± 3.39,P<0.05); butyrylcholinesterase (BchE) (3.22 ± 0.45, 3.66 ± 0.53vs. 7.99 ± 0.79,P<0.05); and acetylcholine (Ach) (4.10 ± 0.38, 3.03 ± 0.25vs.1.72 ± 0.50, P<0.05) significantly increased.Conclusions The GZT volatile oil mainly from Guizhi and Shengjiang can improve the learning and memory ability in dementia mice induced by scopolamine via a cholinergic mechnism.

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