人参-知母-赤芍提取物对血管性痴呆大鼠海马神经元的保护作用及机制
|
摘要
目的:观察人参-知母-赤芍提取物对血管性痴呆大鼠海马N-甲基-D-天冬氨酸受体1(NMDAR1)的影响,探讨其保护海马神经元的作用机制。方法:60只SPF级雄性SD大鼠,随机分为正常组,假手术组,模型组,人参-知母-赤芍提取物组(0. 20 g·kg-1)和美金刚组(2. 1 mg·kg-1),每组12只。采用双侧颈总动脉反复夹闭合并腹腔注射硝普钠的方法建立血管性痴呆大鼠模型,造模后,正常组,假手术组,模型组给予同等体积生理盐水,每日1次,连续14 d。采用Morris水迷宫评价各组大鼠学习记忆能力;苏木素-伊红(HE)染色观察海马CA1区病理改变;蛋白免疫印迹法(Western blot)检测海马神经元胞膜NMDAR1的蛋白表达水平;免疫组化法(IHC)检测海马NMDAR1的表达情况;实时荧光定量PCR(Real-time PCR)检测海马组织中NMDAR1 mRNA的表达水平。结果:与正常组和假手术组比较,模型组大鼠逃避潜伏期显著延长(P <0. 01),在平台所在象限停留时间及穿越平台的次数显著减少(P <0. 01),海马CA1区神经细胞层次减少,排列紊乱、出现核固缩、神经元丢失,NMDAR1蛋白及mRNA表达显著升高(P <0. 01);与模型组比较,人参-知母-赤芍提取物组,美金刚组逃避潜伏期明显缩短(P <0. 05,P <0. 01),在平台所在象限停留时间及穿越平台的次数明显增加(P <0. 05,P <0. 01),海马CA1区神经元数目与形态有明显改善,海马神经元NMDAR1蛋白及NMDAR1 mRNA表达明显降低(P <0. 05)。结论:人参-知母-赤芍提取物能够改善血管性痴呆大鼠的学习记忆能力,减轻海马CA1区神经元的损伤,其机制可能与下调海马神经元NMDAR1的表达有关。
Objective: To observe the effect of extracts from Ginseng Radix et Rhizoma,Anemarrhenae Rhizoma and Paeoniae Radix Rubra on N-methyl-D-aspartate receptors( NMDAR1) in hippocampal neurons in rats with vascular dementia and investigate its possible mechanism. Method: The 60 SPF male rats were randomly divided into normal group,sham-operated group,model group,traditional Chinese medicine group( 0. 20 g·kg-1)and memantine group( 2. 1 mg·kg-1),with 12 rats in each group. The model was established by repeated ischemia-reperfusion combined with intraperitoneal injection of sodium nitroprusside. After modelling,normal group,sham-operated group and model group were dosed the similar volume of normal saline once a day for 14 days. The learning and memory capacity was assessed by Morris water maze; pathologic change in the CA1 district of hippocampus was assessed by hematoxylin-eosin( HE) staining,and the expression level of NMDAR1 in hippocampal neuron membrane protein was detected by Western blot and immunohistochemistry( IHC), the NMDAR1 mRNA in hippocampal tissue was detected by Real-time PCR. Result: Compared with normal and sham-operated group,the latency period was prolonged in model group( P < 0. 01),the time in the platform quadrant and the frequency of crossing the platform were lessened significantly( P < 0. 01),the disorder of the neurons,the decrease of the neuronal number,and the neuronal necrosis and apoptosis were obversed in the CA1 district,expression of membrane NMDAR1 of the hippocampal neuron and NMDAR1 mRNA were increased significantly( P < 0. 01). Compared with model group,the latency period of extracts from Ginseng Radix et Rhizoma,Anemarrhenae Rhizoma and Paeoniae Radix Rubra group and memantine group were shortened significantly( P < 0. 05, P < 0. 01),the time and frequency were increased( P < 0. 05, P < 0. 01),the pathologic change was improved markedly,the protein expression of membrane NMDAR1 in hippocampal neuron and NMDAR1 mRNA were decreased significantly( P < 0. 01). Conclusion: The extracts from Ginseng Radix et Rhizoma,Anemarrhenae Rhizoma and Paeoniae Radix Rubra can improve the learning and memory capacity of rats with vascular dementia,and alleviate the injury in CA1 district of hippocampus. The mechanism may be related to the down-regulation of NMDAR1 expression in hippocampal neurons.
Objective: To observe the effect of extracts from Ginseng Radix et Rhizoma,Anemarrhenae Rhizoma and Paeoniae Radix Rubra on N-methyl-D-aspartate receptors( NMDAR1) in hippocampal neurons in rats with vascular dementia and investigate its possible mechanism. Method: The 60 SPF male rats were randomly divided into normal group,sham-operated group,model group,traditional Chinese medicine group( 0. 20 g·kg-1)and memantine group( 2. 1 mg·kg-1),with 12 rats in each group. The model was established by repeated ischemia-reperfusion combined with intraperitoneal injection of sodium nitroprusside. After modelling,normal group,sham-operated group and model group were dosed the similar volume of normal saline once a day for 14 days. The learning and memory capacity was assessed by Morris water maze; pathologic change in the CA1 district of hippocampus was assessed by hematoxylin-eosin( HE) staining,and the expression level of NMDAR1 in hippocampal neuron membrane protein was detected by Western blot and immunohistochemistry( IHC), the NMDAR1 mRNA in hippocampal tissue was detected by Real-time PCR. Result: Compared with normal and sham-operated group,the latency period was prolonged in model group( P < 0. 01),the time in the platform quadrant and the frequency of crossing the platform were lessened significantly( P < 0. 01),the disorder of the neurons,the decrease of the neuronal number,and the neuronal necrosis and apoptosis were obversed in the CA1 district,expression of membrane NMDAR1 of the hippocampal neuron and NMDAR1 mRNA were increased significantly( P < 0. 01). Compared with model group,the latency period of extracts from Ginseng Radix et Rhizoma,Anemarrhenae Rhizoma and Paeoniae Radix Rubra group and memantine group were shortened significantly( P < 0. 05, P < 0. 01),the time and frequency were increased( P < 0. 05, P < 0. 01),the pathologic change was improved markedly,the protein expression of membrane NMDAR1 in hippocampal neuron and NMDAR1 mRNA were decreased significantly( P < 0. 01). Conclusion: The extracts from Ginseng Radix et Rhizoma,Anemarrhenae Rhizoma and Paeoniae Radix Rubra can improve the learning and memory capacity of rats with vascular dementia,and alleviate the injury in CA1 district of hippocampus. The mechanism may be related to the down-regulation of NMDAR1 expression in hippocampal neurons.
引文
[1] LI N,LIU R J,Dwyer J M,et al. Glutamate N-methylD-aspartate receptor antagonists rapidly reverse behavioral and synaptic deficits caused by chronic stress exposure.[J]. Biol Psychiat,2011,69(8):754-761.
[2]王业梅,胡建鹏,程惠娟.两种中药复方对脑缺血再灌注大鼠脑组织Glu含量及NMDA受体亚单位NR1表达的影响[J].中国民族民间医药,2013,22(12):33-34.
[3]姚国恩,王景周,陈曼娥.血管性痴呆大鼠认知障碍的NMDAR机制研究[J].第三军医大学学报,2002,(12):1408-1410.
[4] Ashby E L,Kierzkowska M, Hull J,et al. Altered expression of human mitochondrial branched chain aminotransferase in dementia with lewy bodies and vascular dementia[J]. Neurochem Res,2017,42(1):306-319.
[5] Iadecola C. The pathobiology of vascular dementia[J].Neuron,2013,80(4):844-866.
[6]刘雪景.补阳还五汤治疗血管性痴呆临床观察[J].北方药学,2017,14(2):97.
[7]蔡颖颖,蒋卫民.中医药干预血管性认知障碍的研究进展[J].中国中药杂志,2017,42(10):1837-1841.
[8]马克信,韩振蕴,马大勇,等.参知健脑胶囊对拟血管性痴呆大鼠模型皮层和海马中Glu和GABA的影响[J].辽宁中医杂志,2016,43(11):2421-2424.
[9]王蕊,杨秦飞,唐一鹏,等.大鼠拟血管性痴呆模型的建立及中药9602防治作用初探[J].北京中医药大学学报,2000,23(5):30-32.
[10]王频,汤敬一,杨骏,等.艾灸对血管性痴呆大鼠海马内VEGF、flt-1、b FGF及b FGF-r表达的影响[J].中国中西医结合杂志,2012,32(1):97-101.
[11]马克信,韩振蕴,张韩瑜嘉,等.参知健脑胶囊对脑缺血大鼠局部脑血流量的影响[J].吉林中医药,2018,38(1):72-75
[12] Thal D R,Grinberg L T,Attems J. Vascular dementia:different forms of vessel disorders contribute to the development of dementia in the elderly brain[J]. Exp Gerontol,2012,47(11):816-824.
[13]王枫,刘敬霞,顾玉宝,等.血管性痴呆的中医研究进展和治疗现状[J].辽宁中医杂志,2016,43(9):1997-2000.
[14] Erkinjuntti T,Román G,Gauthier S,et al. Emerging therapies for vascular dementia and vascular cognitive impairment[J]. Stroke,2004,35(4):1010-1017.
[15]田金洲,王永炎,刘峘,等.血管性痴呆发病机理的研究[J].中医杂志,2003,44(8):565-567.
[16]杨傲然.参知健脑片组分配伍对拟阿尔茨海默病细胞模型的保护作用及机制探讨[D].北京:北京中医药大学,2007.
[17]杨傲然,田昕,钟华,等.参知健脑片组方配伍对Aβ致阿尔茨海默病细胞模型细胞周期和凋亡率的影响[J].中华中医药学刊,2010,28(3):562-564.
[18]牛美英.参知健脑方对缺氧致AD细胞模型线粒体损伤的保护机制的实验研究[D].北京:北京中医药大学,2016.
[19]马大勇,韩振蕴,范吉平,等.参知健脑胶囊对记忆障碍小鼠模型行为学的影响[J].中国医药导报,2012,9(7):32-34.
[20] Foster T C,Kyritsopoulos C,Kumar A. Central role for NMDA receptors in redox mediated impairment of synaptic function during aging and Alzheimer's disease[J]. Behav Brain Res,2017,322(Pt B):223-232.
[21]于文娇,杨巍巍,李昕,等.α-突触核蛋白通过Rab5B下调海马神经元膜表面NMDA受体含量及其介导的Ca~(2+)内流和内向电流[J].首都医科大学学报,2017,38(6):868-873.
[22] CHENG F, LI X, LI Y, et al. Alpha-Synuclein promotes clathrin-mediated NMDA receptor endocytosis and attenuates NMDA-induced dopaminergic cell death[J]. J Neurochem,2011,119(4):815-825.
[23]赵宝敏,张楠,程焱.盐酸美金刚对血管性痴呆大鼠N-甲基-D天冬氨酸受体的影响[J].中华老年心脑血管病杂志,2015,17(3):303-306.
[2]王业梅,胡建鹏,程惠娟.两种中药复方对脑缺血再灌注大鼠脑组织Glu含量及NMDA受体亚单位NR1表达的影响[J].中国民族民间医药,2013,22(12):33-34.
[3]姚国恩,王景周,陈曼娥.血管性痴呆大鼠认知障碍的NMDAR机制研究[J].第三军医大学学报,2002,(12):1408-1410.
[4] Ashby E L,Kierzkowska M, Hull J,et al. Altered expression of human mitochondrial branched chain aminotransferase in dementia with lewy bodies and vascular dementia[J]. Neurochem Res,2017,42(1):306-319.
[5] Iadecola C. The pathobiology of vascular dementia[J].Neuron,2013,80(4):844-866.
[6]刘雪景.补阳还五汤治疗血管性痴呆临床观察[J].北方药学,2017,14(2):97.
[7]蔡颖颖,蒋卫民.中医药干预血管性认知障碍的研究进展[J].中国中药杂志,2017,42(10):1837-1841.
[8]马克信,韩振蕴,马大勇,等.参知健脑胶囊对拟血管性痴呆大鼠模型皮层和海马中Glu和GABA的影响[J].辽宁中医杂志,2016,43(11):2421-2424.
[9]王蕊,杨秦飞,唐一鹏,等.大鼠拟血管性痴呆模型的建立及中药9602防治作用初探[J].北京中医药大学学报,2000,23(5):30-32.
[10]王频,汤敬一,杨骏,等.艾灸对血管性痴呆大鼠海马内VEGF、flt-1、b FGF及b FGF-r表达的影响[J].中国中西医结合杂志,2012,32(1):97-101.
[11]马克信,韩振蕴,张韩瑜嘉,等.参知健脑胶囊对脑缺血大鼠局部脑血流量的影响[J].吉林中医药,2018,38(1):72-75
[12] Thal D R,Grinberg L T,Attems J. Vascular dementia:different forms of vessel disorders contribute to the development of dementia in the elderly brain[J]. Exp Gerontol,2012,47(11):816-824.
[13]王枫,刘敬霞,顾玉宝,等.血管性痴呆的中医研究进展和治疗现状[J].辽宁中医杂志,2016,43(9):1997-2000.
[14] Erkinjuntti T,Román G,Gauthier S,et al. Emerging therapies for vascular dementia and vascular cognitive impairment[J]. Stroke,2004,35(4):1010-1017.
[15]田金洲,王永炎,刘峘,等.血管性痴呆发病机理的研究[J].中医杂志,2003,44(8):565-567.
[16]杨傲然.参知健脑片组分配伍对拟阿尔茨海默病细胞模型的保护作用及机制探讨[D].北京:北京中医药大学,2007.
[17]杨傲然,田昕,钟华,等.参知健脑片组方配伍对Aβ致阿尔茨海默病细胞模型细胞周期和凋亡率的影响[J].中华中医药学刊,2010,28(3):562-564.
[18]牛美英.参知健脑方对缺氧致AD细胞模型线粒体损伤的保护机制的实验研究[D].北京:北京中医药大学,2016.
[19]马大勇,韩振蕴,范吉平,等.参知健脑胶囊对记忆障碍小鼠模型行为学的影响[J].中国医药导报,2012,9(7):32-34.
[20] Foster T C,Kyritsopoulos C,Kumar A. Central role for NMDA receptors in redox mediated impairment of synaptic function during aging and Alzheimer's disease[J]. Behav Brain Res,2017,322(Pt B):223-232.
[21]于文娇,杨巍巍,李昕,等.α-突触核蛋白通过Rab5B下调海马神经元膜表面NMDA受体含量及其介导的Ca~(2+)内流和内向电流[J].首都医科大学学报,2017,38(6):868-873.
[22] CHENG F, LI X, LI Y, et al. Alpha-Synuclein promotes clathrin-mediated NMDA receptor endocytosis and attenuates NMDA-induced dopaminergic cell death[J]. J Neurochem,2011,119(4):815-825.
[23]赵宝敏,张楠,程焱.盐酸美金刚对血管性痴呆大鼠N-甲基-D天冬氨酸受体的影响[J].中华老年心脑血管病杂志,2015,17(3):303-306.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |