天麻多糖对脑瘫幼鼠脑内神经递质的影响
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摘要
目的:探讨天麻多糖(Gastrodiae Rhizoma polysaccharide,GRPS)对脑瘫幼鼠神经递质的影响及其机制。方法:58只SD幼鼠,除空白组外,48只幼鼠结扎左侧颈总动脉并缺氧1 h造脑瘫模型,成模大鼠随机分为模型组,施普善组(2.5 m L·kg-1),GRPS高、低剂量组(300,150 mg·kg-1)。给药21 d后,Y型迷宫及跳台实验检测大鼠记忆力。化学或酶联免疫吸附(ELISA)法检测大脑皮层及左侧海马一氧化氮(nitric oxide,NO),乙酰胆碱脂酶(acetylcholin esterase,ACHE),5-羟色胺(5-hydroxytryptamin,5-HT),去甲肾上腺素(noradrenaline,NE)和γ-氨基丁酸(γ-aminobutyric acid,GABA)水平。实时荧光定量PCR(Real-time PCR)及ELISA法检测大脑皮层及海马内皮型一氧化氮合酶(endothelial nitric oxide synthase,e NOS)表达及含量。苏木素-伊红(HE)染色观察右侧海马组织结构。结果:与空白组比较,模型大鼠行为学错误次数增加,跳台潜伏期减少,大脑皮层和海马中5-HT,NE和GABA下降,ACHE和Glu增加(P<0.05,P<0.01)。与模型组比,GRPS高和低剂量组大鼠行为学错误次数减少,跳台潜伏期增加(P<0.05,P<0.01)。GRPS高和低剂量组大鼠在大脑皮层中NO,NE,5-HT和e NOS增加,ACHE减少;GRPS高剂量组大鼠海马的NO,NE和e NOS增加,ACHE减少(P<0.05,P<0.01)。GRPS低高剂量组大鼠海马组织结构未见水肿,细胞排列整齐。结论:GRPS可以增强脑瘫幼鼠记忆力,其机制与增加大脑皮层及海马中NO,NE和5-HT含量,降低ACHE水平,增加e NOS表达,保护海马区组织有关。
Objective: To analyze the effect of Gastrodiae Rhizoma polysaccharide( GRPS) on neutral transmitter in the immature rats with cerebral palsy. Method: A total of 58 SD immature rats were used in the experiment,and except in the blank control ones,common carotid arteries of all the left 48 rats were ligatured and combined with hypoxia treatment to establish cerebral palsy models in immature rats. The successfully modeled rats were randomly divided into model group,cerebrolysin group( 2. 5 m L·kg-1),GRPS low( 150 mg·kg-1) and high dose( 300 mg·kg-1) groups. After treatment for 21 days,Y maze and diving platform experiment were used to explore the memory ability of rats. The levels of nitric oxide( NO), acetylcholine esterase( ACHE),5-hydroxytryptamin( 5-HT), noradrenaline( NE) and γ-aminobutyric acid( GABA) in cerebral cortex and hippocampus were measured by using ELISA or chemical method. Real-time PCR and ELISA were used to detect the expression levels of endothelial nitric oxide synthase( e NOS) mRNA and protein in brain. HE staining was used to observe the hippocampus tissue structure. Result: As compared with normal blank group,the error number in behavior was significantly increased in model group rats; the step down latency was decreased; and also,the levels of 5-HT, NE and GABA in cerebral cortex and hippocampus of model group rats were significantly decreased,while the levels of ACHE and Glu were increased significantly( P < 0. 05,P < 0. 01). As compared with the model group,the levels of NO,NE,5-HT,and e NOS in cerebral cortex and hippocampus of GRPS low and high dose group rats were increased,while the level of ACHE was decreased( P < 0. 05,P < 0. 01); the levels of NO,NE,and expression of e NOS in hippocampus of GRPS high dose group were increased,while the levels of ACHE were decreased( P < 0. 05,P < 0. 01). The regular cell position was observed in hippocampus of GRPS low and high dose groups,without edema. Conclusion: GRPS could improve the memory of cerebral palsy immature rats,and the mechanism may be associated with increasing NO,ACHE,NE,5-HT and e NOS levels in cerebral cortex and hippocampus,and protection on hippocampus tissue structure of rats.
Objective: To analyze the effect of Gastrodiae Rhizoma polysaccharide( GRPS) on neutral transmitter in the immature rats with cerebral palsy. Method: A total of 58 SD immature rats were used in the experiment,and except in the blank control ones,common carotid arteries of all the left 48 rats were ligatured and combined with hypoxia treatment to establish cerebral palsy models in immature rats. The successfully modeled rats were randomly divided into model group,cerebrolysin group( 2. 5 m L·kg-1),GRPS low( 150 mg·kg-1) and high dose( 300 mg·kg-1) groups. After treatment for 21 days,Y maze and diving platform experiment were used to explore the memory ability of rats. The levels of nitric oxide( NO), acetylcholine esterase( ACHE),5-hydroxytryptamin( 5-HT), noradrenaline( NE) and γ-aminobutyric acid( GABA) in cerebral cortex and hippocampus were measured by using ELISA or chemical method. Real-time PCR and ELISA were used to detect the expression levels of endothelial nitric oxide synthase( e NOS) mRNA and protein in brain. HE staining was used to observe the hippocampus tissue structure. Result: As compared with normal blank group,the error number in behavior was significantly increased in model group rats; the step down latency was decreased; and also,the levels of 5-HT, NE and GABA in cerebral cortex and hippocampus of model group rats were significantly decreased,while the levels of ACHE and Glu were increased significantly( P < 0. 05,P < 0. 01). As compared with the model group,the levels of NO,NE,5-HT,and e NOS in cerebral cortex and hippocampus of GRPS low and high dose group rats were increased,while the level of ACHE was decreased( P < 0. 05,P < 0. 01); the levels of NO,NE,and expression of e NOS in hippocampus of GRPS high dose group were increased,while the levels of ACHE were decreased( P < 0. 05,P < 0. 01). The regular cell position was observed in hippocampus of GRPS low and high dose groups,without edema. Conclusion: GRPS could improve the memory of cerebral palsy immature rats,and the mechanism may be associated with increasing NO,ACHE,NE,5-HT and e NOS levels in cerebral cortex and hippocampus,and protection on hippocampus tissue structure of rats.
引文
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[2]梁海英.儿童脑瘫综合治疗进展[J].中国医院用药评价与分析,2016,16(1):232-233.
[3]徐鹏,赵毅,张新.脑瘫患儿脑脊液氨基酸类神经递质的变化[J].吉林大学学报:医学版,2004,30(1):117-119.
[4]XU D Q,ZHOU J J,LIU Y H.Research the extraction and detection method of gastrodin content in Gastrodia elata[J].J Chin Medl Mater,2012,35(11):1799-1804.
[5]黄锐,赵健,吴锋,等.电针联合天麻多糖对脑缺血大鼠基底外侧杏仁核巢蛋白、脑源性神经营养因子表达的影响[J].针刺研究,2016,41(3):230-234.
[6]任守利,刘塔斯,刘宇婧,等.天麻多糖提取工艺的研究[J].湖南中医药大学学报,2010,30(1):37-40.
[7]Rumajogee P,Bregman T,Miller S P,et al.Rodent hypoxia-ischemia models for cerebral palsy research:asystematic review[J].Front Neurol,2016,doi:10.3389/fneur.2016.00057.
[8]王丽敏,郭佳丽,杨自金,等.人脐血间充质干细胞移植与神经节苷脂注射治疗脑瘫大鼠的对比研究[J].中国康复,2015,30(5):331-334.
[9]杜鹏.针灸配合康复训练治疗小儿脑瘫30例临床观察[J].中医儿科杂志,2014,10(3):57-59.
[10]毛勇,苏敏,那敏,等.天麻粉喂养大鼠30d毒性试验[J].中国药理学与毒理学杂志,2013,27(3):561.
[11]于滨,左增艳,孔维佳.天麻细粉片毒性及安全性的实验研究[J].中国当代医药,2014,21(21):6-10.
[12]Bird C M.The role of the hippocampus in recognition memory[J].Cortex,2017,1(93):155-165.
[13]Witt J A,Helmstaedter C,Elger C E.Is there evidence of a subordinate role of the hippocampal CA1 field for declarative memory formation?[J].Brain,2015,138(4):e343.
[14]Gautam A,Wadhwa R,Thakur M K,et al.Assessment of cholinergic properties of ashwagandha leaf-extract in the amnesic mouse brain[J].Ann Neurosci,2016,23(2):68-75.
[15]Sundaramurthy S,Annamalai B,Samuvel D J,et al.Modulation of serotonin transporter function by kappaopioid receptor ligands[J].Neuropharmacology,2017,113(1):281-292.
[16]孔繁一,刘诗翔,张皓,等.脑内谷氨酸和5-羟色胺增高是高原睡眠障碍的独立预测因素[J].疑难病杂志,2013,12(11):843-847.
[17]卢桃利,罗勇,孙宏毅,等.电针对局灶脑缺血再灌注大鼠大脑皮质e NOS mRNA及蛋白、MMP-9蛋白表达的影响[J].基础医学与临床,2012,32(2):158-163.