益智仁挥发油对实验性帕金森病的作用研究
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摘要
帕金森病(Parkinson's Disease,PD)是一种老年神经系统退行性疾病,PD患病率呈逐年上升趋势,在老年退行性疾病中发病率位居第二,仅次于阿尔茨海默病,发病率已达1.03%。目前临床所用的治疗PD药物数量少,且均有长期用药后疗效下降并出现严重不良反应等缺点,因此,开发高效低毒的PD防治药物显得尤为迫切。
益智仁为姜科类植物益智(Alpinia Oxyphylla Miq.)的成熟干燥果实,是我国著名的四大南药之一。为了加快益智药材的开发,我国药学工作者对其进行了较系统的研究。研究发现,益智仁主要化学成分为挥发油类、微量元素、萜类、黄酮类及庚烷类等,其中挥发油占1%~2%,研究益智仁化学成分的重点为挥发油成分,用气质联用法已经分析出100余种化学成分,含量较高的有聚伞花烃、香橙醛、芳樟醇、桃金娘醛、α-蒎烯、β-蒎烯等,还含有锌、铜、铁、锰、钴、镁、钙等微量元素,其中锌、锰的含量远远高于其他补益药。现代医学研究表明,益智仁具有强心、抗癌、抗过敏、抗衰老、镇静、镇痛等多方面药理作用。近年来,以益智仁为主的方剂在益智健脑方面的临床应用越来越多,并且在动物实验中进一步证实益智仁具有良好的脑保护作用,可用于老年性痴呆及其他智能障碍类疾病的治疗,对神经细胞有抗氧化、抗衰老、减轻兴奋性毒性和抗凋亡作用。
鉴于益智主产于海南岛,且现已大面积栽培,产量高、品质好。因此,充分利用益智进行深加工,研究益智仁挥发油对PD的作用,开发益智仁挥发油作为PD治疗药物很有必要,亦有特色。
目的:研究益智仁挥发油对实验性PD的作用,并初步研究其作用机制。
方法:(1)水蒸气法提取益智仁挥发油,采用气相色谱法分析实验过程中所用益智仁挥发油的化学成分,研究其可控性及稳定性;(2)采用小鼠BLISS法,对益智仁挥发油进行急性毒性研究,测定其LD50;(3)1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methyl-4-phenyl- 1,2,3,6- tetrahydropyri-dine,MPTP)腹腔注射建立C57BL小鼠PD模型,对PD小鼠进行游泳、自主活动、水迷宫行为学实验,观察益智仁挥发油对MPTP所诱导PD的保护作用;(4)建立PD小鼠模型,溶剂提取法分离小鼠纹状体内DA、5-HT,层析法分离小鼠纹状体内HVA,荧光分光光度计法测定DA、5-HT、HVA含量,观察益智仁挥发油对小鼠脑内单胺类神经递质的保护作用;(5)紫外分光光度计法测定PD小鼠纹状体内SOD活力,MDA、GSH含量;(6)采用免疫组化法,分析益智仁挥发油对小鼠黑质致密部神经细胞TH、BCL-2及CASPASE-3蛋白表达的影响,采用TUNEL细胞凋亡技术分析益智仁对小鼠黑质致密部神经细胞凋亡的影响。
结果:(1)益智仁挥发油主要化学成分稳定,质量可控;(2)BLISS法测得益智仁挥发油LD50为8.327ml·kg-1;(3)行为学实验结果显示益智仁挥发油能对抗MPTP所诱导的PD小鼠游泳时间缩短,并且缩短了小鼠游出水迷宫的潜伏期和错误次数,但对小鼠自主活动减少无明显影响;(4)预先给予益智仁挥发油可明显抑制MPTP所引起的小鼠脑组织内GSH含量下降和MDA生成增加,且能抑制纹状体SOD活力的下降;(5)益智仁挥发油能阻遏PD小鼠纹状体内单胺类神经递质下降,预先给予益智仁挥发油可明显抑制小鼠脑内DA、HVA、5-HT含量降低;(6)益智仁挥发油可增加PD小鼠黑质致密部(SNc)TH、BCL-2蛋白的表达,降低SNc内CASPASE-3表达,同时能抑制PD小鼠黑质致密部神经元细胞的凋亡。
结论:(1)益智仁挥发油成分稳定、可控;(2)益智仁挥发油毒性甚微,发现益智仁挥发油能对抗PD所致的小鼠行为学改变;(3)通过对益智仁挥发油抗实验性帕金森病机制的初步研究,发现其抗PD机制可能与下列因素有关:
①增强对氧自由基的清除能力,抑制脂质过氧化反应;
②抑制MAO-B活性,阻遏PD小鼠脑内DA、HVA、5-HT分解代谢;
③益智仁挥发油能增加自身抗氧能力,调节与细胞凋亡密切相关的BCL-2、CASPASE-3、TH等调控蛋白的含量和表达,抑制细胞凋亡。
Parkinson's Disease(PD) is a kind of gerontism nervous system degenerative disease.PD attack rate goes up year by year, which is second only to Alzheimer disease, and its disease incidence has already reached 1.03%.At present,there are few clinical drugs for PD treatment and there are shortcomings,such as curative effect decline and serious adverse effects after long-term administration,it is thus evident that it is important to development high efficiency low toxical drug for PD’s prevention and cure.
Alpinia Oxyphylla Miq.fruit is mature and dry fruit, which is one of the most famous traditional medicine in South-China.To speed up the development of Alpinia Oxyphylla Miq.,Chinese pharmaceutical researchers have undertook systematic researches on AOF.Recent study results indicated that the main chemical compositions in Alpinia Oxyphylla Miq. fruit were essential oil, microelement, terpene flavonoids, heptanes, etc,and essential oil accounted for 1%～2%. As a result, the keypoint to research the chemical composition is to research essential oil.More than 100 kinds of chemical compositions has been analyzed by Gas chromatography mass spectrometry(GC-MS) method.The main compositions were p-cymene, neral,coriandrol, myrtenal,a-pinene,β-pinene, etc,and the main microele- ments were Zn,Cu,Fe,Mn,Gu,Mg,Ca etc.Zn and Mn were obviously higher than other tonic.Modern medicine study has shown that Alpinia Oxyphylla Miq.fruit have the pharmacologic action on strengthen heart power, anti- cancer,anti-anapyhlaxis, anti-aging, cons- tious-sedation and relief pain. Recent years, more and more Alpinia Oxyphylla Miq.fruit as primary composition in some compound preparation has been applicated on clinical researches for intelligence and brain.It has been proved that Alpinia Oxyphylla Miq.fruit has good effects on brain protection in animal experiment, and can be used to treat senile dementia and other disturbance of intelligence. It also has antioxygen, anti-ageing, anti-apoptosis effects and can lessen excitability toxicity effect on nerve cell.
As far as it is concerned that Alpinia Oxyphylla Miq. fruit is mainly produced in Hainan Island, where Alpinia Oxyphylla Miq.has been cultivated in large areas with high output and fine quality, it is feasible to extract essential oil from AOF for PD treatment.
Objective:To study the effects and mechanisms of AOF on the experimental PD.
Methods:(1)Wet distillation method was used to extract Essential oil from AOF;vapor-phase chromatography was applied to analyse the p-Cymene of essential oil extracted from AOF for studying the stability and normallability of essential oil extracted from AOF; (2)BLISS method was used to study acute toxicity test in mice and determine the LD50 ;(3) The C57BL mice were formed with intraperitoneal injections of 1-methyl- 4-phenyl-1,2,3,6-tetrahydropyri-dine(MPTP) to establish PD model; behavioral exhibition was explored by swimming test,water maze experiment and antonomic activities counting experiment;(4)Mice were formed with intraperitoneal injections of MPTP to establish PD model; the dopamine(DA), 5-hydroxytryptamine(5-HT) were abstracted by solvent extraction method; homovanillic acid(HVA) was abstracted by chroma- tographic analysis antigenic method;the DA,5-HT,HVA levels of striata were analysed by Fluorospectraphotometry method;observe essential oil extracted from AOF effect on monoamine neurotransmitter in mice brain; (5)The malondialdehyde(MDA),reduced glutathione hormone(GSH) and superoxide dismutase (SOD) activity from mice striatum were measured by ultraviolet spectrophotometer method to explore the mechanism of protect- ive effects;(6)The tyrosine hydroxylase(TH),B-cell leukemia- lymphoma- 2(BCL-2)and CASPASE-3 expressed in SNc were detected by Immunohisto- chemical SABC method;TUNEL staining method was used to detecte apoptosis in SNc of mice.
Results:(1)The chemical compositions of essential oil extracted from AOF were stable and normallable; (2)The LD50 of essential oil extracted from AOF was 8.3269 ml·kg-1 and its 95% confident limit was 7.037-10.06 ml·kg-1,which suggests the toxicity of essential oil extracted from AOF might be minor;(3)Ethology experimental results showed that the AOF groups had significant difference with the MPTP model group;essential oil extracted from AOF could prolong swimming time of PD mice,shorten latency phase of mice in the water maze and decrease frequency of mice into blind-ending , but hadn’t significant effect on autonomic activities of PD mice;(4)Essential oil extracted from AOF obviously antagonized MPTP-induced elevation of MDA,as well as decreased of GSH level and SOD activity of brain tissue; (5)Essential oil extracted from AOF obviously antagonize MPTP-induced decrease of DA,HVA,5-HT of PD mice; (6)Essential oil extracted from AOF could significantly increase the TH,BCL-2 expression,obviously decrease CASPASE-3expression in SNc, meanwhile inhibit nerve cell Apoptosis in SNc.
Conclusion:(1)The compositions of essential oil extracted from AOF are stable and normallable;(2)The toxicity of essential oil extracted from AOF is very low;essential oil extracted from AOF could antagonize ethology change of PD mice induced by MPTP,which suggests that AOF can have effects on prevention and cure PD induced by MPTP.(3)It is showed that the anti-Parkinson's disease mechanisms of essential oil extracted from AOF might relate to the following factors.
①Essential oil extracted from AOF inhibits generation of oxygen- derived free radicals,and accelerates cleaning of free radicals,and relieve lipid peroxidation reaction, and maintains contents of SOD, GSH in normal level.
②Essential oil extracted from AOF inhibits activity of monoamine oxidase-B(MAO-B), repress DA, 5-HT destructive metabolism.
③Essential oil extracted from AOF could increase self-antioxygen capability, regulate the expression of modulin that intimate related to apoptosis such as BCL-2、CASEPASE-3、TH,etc.,then inhibits apoptosis.
益智仁为姜科类植物益智(Alpinia Oxyphylla Miq.)的成熟干燥果实,是我国著名的四大南药之一。为了加快益智药材的开发,我国药学工作者对其进行了较系统的研究。研究发现,益智仁主要化学成分为挥发油类、微量元素、萜类、黄酮类及庚烷类等,其中挥发油占1%~2%,研究益智仁化学成分的重点为挥发油成分,用气质联用法已经分析出100余种化学成分,含量较高的有聚伞花烃、香橙醛、芳樟醇、桃金娘醛、α-蒎烯、β-蒎烯等,还含有锌、铜、铁、锰、钴、镁、钙等微量元素,其中锌、锰的含量远远高于其他补益药。现代医学研究表明,益智仁具有强心、抗癌、抗过敏、抗衰老、镇静、镇痛等多方面药理作用。近年来,以益智仁为主的方剂在益智健脑方面的临床应用越来越多,并且在动物实验中进一步证实益智仁具有良好的脑保护作用,可用于老年性痴呆及其他智能障碍类疾病的治疗,对神经细胞有抗氧化、抗衰老、减轻兴奋性毒性和抗凋亡作用。
鉴于益智主产于海南岛,且现已大面积栽培,产量高、品质好。因此,充分利用益智进行深加工,研究益智仁挥发油对PD的作用,开发益智仁挥发油作为PD治疗药物很有必要,亦有特色。
目的:研究益智仁挥发油对实验性PD的作用,并初步研究其作用机制。
方法:(1)水蒸气法提取益智仁挥发油,采用气相色谱法分析实验过程中所用益智仁挥发油的化学成分,研究其可控性及稳定性;(2)采用小鼠BLISS法,对益智仁挥发油进行急性毒性研究,测定其LD50;(3)1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methyl-4-phenyl- 1,2,3,6- tetrahydropyri-dine,MPTP)腹腔注射建立C57BL小鼠PD模型,对PD小鼠进行游泳、自主活动、水迷宫行为学实验,观察益智仁挥发油对MPTP所诱导PD的保护作用;(4)建立PD小鼠模型,溶剂提取法分离小鼠纹状体内DA、5-HT,层析法分离小鼠纹状体内HVA,荧光分光光度计法测定DA、5-HT、HVA含量,观察益智仁挥发油对小鼠脑内单胺类神经递质的保护作用;(5)紫外分光光度计法测定PD小鼠纹状体内SOD活力,MDA、GSH含量;(6)采用免疫组化法,分析益智仁挥发油对小鼠黑质致密部神经细胞TH、BCL-2及CASPASE-3蛋白表达的影响,采用TUNEL细胞凋亡技术分析益智仁对小鼠黑质致密部神经细胞凋亡的影响。
结果:(1)益智仁挥发油主要化学成分稳定,质量可控;(2)BLISS法测得益智仁挥发油LD50为8.327ml·kg-1;(3)行为学实验结果显示益智仁挥发油能对抗MPTP所诱导的PD小鼠游泳时间缩短,并且缩短了小鼠游出水迷宫的潜伏期和错误次数,但对小鼠自主活动减少无明显影响;(4)预先给予益智仁挥发油可明显抑制MPTP所引起的小鼠脑组织内GSH含量下降和MDA生成增加,且能抑制纹状体SOD活力的下降;(5)益智仁挥发油能阻遏PD小鼠纹状体内单胺类神经递质下降,预先给予益智仁挥发油可明显抑制小鼠脑内DA、HVA、5-HT含量降低;(6)益智仁挥发油可增加PD小鼠黑质致密部(SNc)TH、BCL-2蛋白的表达,降低SNc内CASPASE-3表达,同时能抑制PD小鼠黑质致密部神经元细胞的凋亡。
结论:(1)益智仁挥发油成分稳定、可控;(2)益智仁挥发油毒性甚微,发现益智仁挥发油能对抗PD所致的小鼠行为学改变;(3)通过对益智仁挥发油抗实验性帕金森病机制的初步研究,发现其抗PD机制可能与下列因素有关:
①增强对氧自由基的清除能力,抑制脂质过氧化反应;
②抑制MAO-B活性,阻遏PD小鼠脑内DA、HVA、5-HT分解代谢;
③益智仁挥发油能增加自身抗氧能力,调节与细胞凋亡密切相关的BCL-2、CASPASE-3、TH等调控蛋白的含量和表达,抑制细胞凋亡。
Parkinson's Disease(PD) is a kind of gerontism nervous system degenerative disease.PD attack rate goes up year by year, which is second only to Alzheimer disease, and its disease incidence has already reached 1.03%.At present,there are few clinical drugs for PD treatment and there are shortcomings,such as curative effect decline and serious adverse effects after long-term administration,it is thus evident that it is important to development high efficiency low toxical drug for PD’s prevention and cure.
Alpinia Oxyphylla Miq.fruit is mature and dry fruit, which is one of the most famous traditional medicine in South-China.To speed up the development of Alpinia Oxyphylla Miq.,Chinese pharmaceutical researchers have undertook systematic researches on AOF.Recent study results indicated that the main chemical compositions in Alpinia Oxyphylla Miq. fruit were essential oil, microelement, terpene flavonoids, heptanes, etc,and essential oil accounted for 1%～2%. As a result, the keypoint to research the chemical composition is to research essential oil.More than 100 kinds of chemical compositions has been analyzed by Gas chromatography mass spectrometry(GC-MS) method.The main compositions were p-cymene, neral,coriandrol, myrtenal,a-pinene,β-pinene, etc,and the main microele- ments were Zn,Cu,Fe,Mn,Gu,Mg,Ca etc.Zn and Mn were obviously higher than other tonic.Modern medicine study has shown that Alpinia Oxyphylla Miq.fruit have the pharmacologic action on strengthen heart power, anti- cancer,anti-anapyhlaxis, anti-aging, cons- tious-sedation and relief pain. Recent years, more and more Alpinia Oxyphylla Miq.fruit as primary composition in some compound preparation has been applicated on clinical researches for intelligence and brain.It has been proved that Alpinia Oxyphylla Miq.fruit has good effects on brain protection in animal experiment, and can be used to treat senile dementia and other disturbance of intelligence. It also has antioxygen, anti-ageing, anti-apoptosis effects and can lessen excitability toxicity effect on nerve cell.
As far as it is concerned that Alpinia Oxyphylla Miq. fruit is mainly produced in Hainan Island, where Alpinia Oxyphylla Miq.has been cultivated in large areas with high output and fine quality, it is feasible to extract essential oil from AOF for PD treatment.
Objective:To study the effects and mechanisms of AOF on the experimental PD.
Methods:(1)Wet distillation method was used to extract Essential oil from AOF;vapor-phase chromatography was applied to analyse the p-Cymene of essential oil extracted from AOF for studying the stability and normallability of essential oil extracted from AOF; (2)BLISS method was used to study acute toxicity test in mice and determine the LD50 ;(3) The C57BL mice were formed with intraperitoneal injections of 1-methyl- 4-phenyl-1,2,3,6-tetrahydropyri-dine(MPTP) to establish PD model; behavioral exhibition was explored by swimming test,water maze experiment and antonomic activities counting experiment;(4)Mice were formed with intraperitoneal injections of MPTP to establish PD model; the dopamine(DA), 5-hydroxytryptamine(5-HT) were abstracted by solvent extraction method; homovanillic acid(HVA) was abstracted by chroma- tographic analysis antigenic method;the DA,5-HT,HVA levels of striata were analysed by Fluorospectraphotometry method;observe essential oil extracted from AOF effect on monoamine neurotransmitter in mice brain; (5)The malondialdehyde(MDA),reduced glutathione hormone(GSH) and superoxide dismutase (SOD) activity from mice striatum were measured by ultraviolet spectrophotometer method to explore the mechanism of protect- ive effects;(6)The tyrosine hydroxylase(TH),B-cell leukemia- lymphoma- 2(BCL-2)and CASPASE-3 expressed in SNc were detected by Immunohisto- chemical SABC method;TUNEL staining method was used to detecte apoptosis in SNc of mice.
Results:(1)The chemical compositions of essential oil extracted from AOF were stable and normallable; (2)The LD50 of essential oil extracted from AOF was 8.3269 ml·kg-1 and its 95% confident limit was 7.037-10.06 ml·kg-1,which suggests the toxicity of essential oil extracted from AOF might be minor;(3)Ethology experimental results showed that the AOF groups had significant difference with the MPTP model group;essential oil extracted from AOF could prolong swimming time of PD mice,shorten latency phase of mice in the water maze and decrease frequency of mice into blind-ending , but hadn’t significant effect on autonomic activities of PD mice;(4)Essential oil extracted from AOF obviously antagonized MPTP-induced elevation of MDA,as well as decreased of GSH level and SOD activity of brain tissue; (5)Essential oil extracted from AOF obviously antagonize MPTP-induced decrease of DA,HVA,5-HT of PD mice; (6)Essential oil extracted from AOF could significantly increase the TH,BCL-2 expression,obviously decrease CASPASE-3expression in SNc, meanwhile inhibit nerve cell Apoptosis in SNc.
Conclusion:(1)The compositions of essential oil extracted from AOF are stable and normallable;(2)The toxicity of essential oil extracted from AOF is very low;essential oil extracted from AOF could antagonize ethology change of PD mice induced by MPTP,which suggests that AOF can have effects on prevention and cure PD induced by MPTP.(3)It is showed that the anti-Parkinson's disease mechanisms of essential oil extracted from AOF might relate to the following factors.
①Essential oil extracted from AOF inhibits generation of oxygen- derived free radicals,and accelerates cleaning of free radicals,and relieve lipid peroxidation reaction, and maintains contents of SOD, GSH in normal level.
②Essential oil extracted from AOF inhibits activity of monoamine oxidase-B(MAO-B), repress DA, 5-HT destructive metabolism.
③Essential oil extracted from AOF could increase self-antioxygen capability, regulate the expression of modulin that intimate related to apoptosis such as BCL-2、CASEPASE-3、TH,etc.,then inhibits apoptosis.
引文
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[14] Yu XY,ZHAO H,XU ZQ.Neuroprotective efect of Alpinia oxyphylla fruits extract against glutamate excitotoxiclty in cuhured mouse cortial neurons [J].Neurosci.Res. Commun.2003,33:105-113.
[15] 阳辛风,利美莲.益智与益智洒抗氧化活性的研究[J].华南热带农业大学学报. 2001,(3):20.
[16] 易美华,薛献明,肖红,等.益钳提取物对油脂抗氧化作用研究[J].海南大学学报. 2002,(1):28.
[1] 李啸.益智仁对多次裸腹蚤的生物学效应:一种延缓衰老药物的筛选实验[J].生物学杂志.2005,22:3-5.
[2] Yu XY,ZHAO H,XU ZQ.Neuroprotective efect of Alpinia oxyphylla fruits extract against glutamate excitotoxiclty in cuhured mouse cortial neurons[J].Neurosci. Res. Commun.2003,33:105-113.
[3] 阳辛风,利美莲.益智与益智洒抗氧化活性的研究[J].华南热带农业大学学报.2001,(3):20.
[4] 中国药典.一部.2005:附录 ΧD57-58.
[5] 纪明慧,刘红,何猛雄等.益智挥发油的提取及包合工艺的研究[J].中成药.2005, 27(11):1336-1338.
[6] 徐淑云,卞如濂,陈修.药理学实验方法学,第三版(M).北京人民卫生出版社.2002.201-210,713-735,693-695.
[7] Donnan GA,Willis GL,Kaczmarczyk SJ.et al.Motor function in the 1-methyl- 4- Phenyl-1,2,3,6-tetrahydropyridine-treated mouse[J].Neurol Sci.1987,77(2-3):185- 191.
[8] Kawai H,Makino Y,Hirobe M,et a1.Novel endogenous 1,2,3,4- tetrahydroiso- quinoline derivatives:uPtake by doPamine transPorter and activity to induce Parkin- sonism[J].Neurochem.l998,70(2):745-751.
[9] 林敬明,贺巍,吴忠等.益智挥发油成分的GC-MS分析[J].中药材.2000,23(8):448.
[10] 罗秀珍,余竞光,徐丽珍等.中药益智挥发油化学成分[J].中国中药杂志.2001,(4): 262-265.
[11] 张如意,张兰,叶翠飞等.MPTP 对小鼠行为学及脑纹状体多巴胺含量的影响[J].中国行为医学科学杂志.2001,10(3):164-166.
[12] 钟恒亮,王荔萍,陈力.益智口服液镇静催眠作用实验研究[J].贵阳医学院学报.2002,(2):l32-134.
[13] Taylor AE,Saint-Cyr JA.The neuropsychology of Parkinson ’s disease[J].Brain- Cogn.1995,28(3):281-296.
[14] 乔晋,屈秋民,曹红梅等.帕金森病患者智能损害及其危险因素探讨[J].中国临床康复.2002,6(7):942-943.
[15] Wang L,Cheng ZH,Li HH,et al.Laboratory memory an d everyday memory in patients with Parkinsons’s disease[J].Zhongguo Lingchuang Kangfu.2002,6(23):3606-3607(China).
[1] Dauer W,Przedborski S.Parkinson’s disease:mechanisms and models[J]. Neuron. 2003,39:889-909.
[2] 戚辰,刘振国.帕金森病发病机制研究进展:a-synuclein与氧化应激[J].中国神经医学杂志.2004,20(4):327-331.
[3] Schapira AH,Gu M,Taanman JW,et a1.Mitochondria in the etiology and pathogen- nesis of Parkinson’s disease[J].Ann Neurol,1998, 44(Suppl 1):s89-s98.
[4] Hashimoto M,Hsu L J,Xia Y,et a1.Oxidative stress induces Amyloid-like aggregate formation of NACP/alpha-synuclein in vitro[J].Neuroreport. 1999,10:717-721.
[5] 徐叔云.药理实验方法学[M].第 2 版.北京:人民卫生出版社.1991,440-446.
[6] 周宜灿,陈晓眷,朱元贵,等.N-乙酰半胱氨酸防止帕金森病鼠黑质神经元凋亡[J].中国神经科学杂志.2003 Nov,19(6):384-388.
[7] 赵磊,蒲小平.类叶升麻苷对MPTP所致帕金森病小鼠模型的神经保护作用[J].中国药理学通报.2007,23(1) :42-46.
[8] 郭续文.利福平保护MPTP所致PD小鼠中脑多巴胺能神经元凋亡的实验研究[J].中华神经医学杂志.2006,6(5):546-549.
[9] Often D,Beart PM,Cheung NS,et a1.Transgenic mice expressing human bel-2 in their neurons are resistant to 6-hydroxydopamine and 1-methyl-4-phenyl-1,2,3,6- tetrahydropyridine neurotoxicity[J].Proc Natl Acad Sci USA.1998,95(10):5789- 5789.
[10] 丁正同,任惠民,蒋雨平等.百草枯对小鼠黑质纹状体多巴胺能系统的影响[J].复旦学报.2001,Jan 28(1):28-31.
[11] Holthoff VA,Vieregge P,Kessler J,et al.Discordant twins with Parkinson’disease:positron emission tomography and early signs of impaired cognitive circuits[J]. Ann Neurol.1994,36(2):176-182.
[12] Yoshino H,Hattori Y,ImaiH,et al.Sparteine oxidation by hepatic Cytochrome P-450 in patients with Parkinson’s disease[J].RinshoShinkeigaku.1993,33(3): 261-265.
[13] Iacopino AM,Christakos S.Specific reduction of calcium-binding protein (28- kilodaltm calbindin-D)gene expression in aging and neurodegenerative diseases [J].Proc Natl Acad Sci USA.1990,87(11):4078-4082.
[14] Gerlach M,Riederer P,Przuntek H,et a1.MPTP mechanisms of neurotoxicity and their implications for Parkinson’s disease[J].Eur J Pharmacol.1991,208(4):273- 286.
[15] 张如意,张兰,叶翠飞等.MPTP 对小鼠行为学及脑纹状体多巴胺含量的影响[J].中国行为医学科学杂志.2001,10(3):164-166.
[16] Cohen G,Fahn S.The oxidant stress hypothesis in Parkinson’s disease: Evidence supporting it[J].Ann Neurol.1992,32:804-8l2.
[17] Zhang P,Land W,Lee S,et alElectron tomography of degenerating neurons in micewith abnormal regulation of iron metabolism[J].J Struct Biol.2005, 150(2): 144- 153.
[18] Jenner P.Oxidative stress in Parkinson’s disease[J].Ann Neurol.2003, 53(suppl3): S26-36.
[19] Bilsland J,Roy S,Xanthoudakis S,et a1.CASPASE inhibitors attenuate 1-methyl-4- phenylpyridinium toxicity in primary cultures of mesencephatic dopaminergic neurons[J].Neurosci.2002,22(7):2637-2649.
[20] Xu Z,Cawthon D,McCastlain K A,et a1.Selective alterations of gene expression in mice induced by MPTP[J].Synapse.2005,55:45-51.
[21] Kingsbury AE,Mardsen CD,Foster OJ.DNA fragmentation in human substantia nigra:apeptosis or perimortem effect?[J].Mov Disord.1998,13(6):877-884.
[1] 尹琳琳,朱兴族.帕金森病的发病机制及药物治疗研究进展[J].国外医学药学分册.2006,33(2):93-96.
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[18] Kanda T,Jackson MJ,Smith L,et al.Combined use of the adenosine A2A antagonist KW-6002 with L-DOPA or with selective D1 or D2 dopamine agonists increases antiparkinsonian activity but not Oyskinesia in MPTP treated monkeys[J]. Exp Neural. 2000,162(2):321-325.
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[20] 牛平,王耀山,吕永利,何祥. 脑源性神经营养因子对帕金森病大鼠黑质多巴胺能神经元的影响[J].中风与神经疾病杂志.2000,17(4):209-211.
[21] 徐诽诽,刘纯青,马涛,周晓棉,曹颖林,张万琴.人参皂苷Re对MPTP致帕金病模型小鼠多巴胺能神经元的保护作用[J].沈阳药科大学学报.2005,22(1):36-42.
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[11] 李玺,王建军,乔成林,等.脑尔康对小鼠学习记忆障碍的改善作用[J].西安医科大学学报.1999,1:20-21.
[12] ]嵇志红,于新宇,张晓利,等.益智仁水提物对东莨菪碱所致记忆获得障碍大鼠的干预效应[J].中国临床康复.2005,9(28):120-122.
[13] 李啸.益智仁对多次裸腹蚤的生物学效应:一种延缓衰老药物的筛选实验[J].生物学杂志.2005,22:3-5.
[14] Yu XY,ZHAO H,XU ZQ.Neuroprotective efect of Alpinia oxyphylla fruits extract against glutamate excitotoxiclty in cuhured mouse cortial neurons [J].Neurosci.Res. Commun.2003,33:105-113.
[15] 阳辛风,利美莲.益智与益智洒抗氧化活性的研究[J].华南热带农业大学学报. 2001,(3):20.
[16] 易美华,薛献明,肖红,等.益钳提取物对油脂抗氧化作用研究[J].海南大学学报. 2002,(1):28.
[1] 李啸.益智仁对多次裸腹蚤的生物学效应:一种延缓衰老药物的筛选实验[J].生物学杂志.2005,22:3-5.
[2] Yu XY,ZHAO H,XU ZQ.Neuroprotective efect of Alpinia oxyphylla fruits extract against glutamate excitotoxiclty in cuhured mouse cortial neurons[J].Neurosci. Res. Commun.2003,33:105-113.
[3] 阳辛风,利美莲.益智与益智洒抗氧化活性的研究[J].华南热带农业大学学报.2001,(3):20.
[4] 中国药典.一部.2005:附录 ΧD57-58.
[5] 纪明慧,刘红,何猛雄等.益智挥发油的提取及包合工艺的研究[J].中成药.2005, 27(11):1336-1338.
[6] 徐淑云,卞如濂,陈修.药理学实验方法学,第三版(M).北京人民卫生出版社.2002.201-210,713-735,693-695.
[7] Donnan GA,Willis GL,Kaczmarczyk SJ.et al.Motor function in the 1-methyl- 4- Phenyl-1,2,3,6-tetrahydropyridine-treated mouse[J].Neurol Sci.1987,77(2-3):185- 191.
[8] Kawai H,Makino Y,Hirobe M,et a1.Novel endogenous 1,2,3,4- tetrahydroiso- quinoline derivatives:uPtake by doPamine transPorter and activity to induce Parkin- sonism[J].Neurochem.l998,70(2):745-751.
[9] 林敬明,贺巍,吴忠等.益智挥发油成分的GC-MS分析[J].中药材.2000,23(8):448.
[10] 罗秀珍,余竞光,徐丽珍等.中药益智挥发油化学成分[J].中国中药杂志.2001,(4): 262-265.
[11] 张如意,张兰,叶翠飞等.MPTP 对小鼠行为学及脑纹状体多巴胺含量的影响[J].中国行为医学科学杂志.2001,10(3):164-166.
[12] 钟恒亮,王荔萍,陈力.益智口服液镇静催眠作用实验研究[J].贵阳医学院学报.2002,(2):l32-134.
[13] Taylor AE,Saint-Cyr JA.The neuropsychology of Parkinson ’s disease[J].Brain- Cogn.1995,28(3):281-296.
[14] 乔晋,屈秋民,曹红梅等.帕金森病患者智能损害及其危险因素探讨[J].中国临床康复.2002,6(7):942-943.
[15] Wang L,Cheng ZH,Li HH,et al.Laboratory memory an d everyday memory in patients with Parkinsons’s disease[J].Zhongguo Lingchuang Kangfu.2002,6(23):3606-3607(China).
[1] Dauer W,Przedborski S.Parkinson’s disease:mechanisms and models[J]. Neuron. 2003,39:889-909.
[2] 戚辰,刘振国.帕金森病发病机制研究进展:a-synuclein与氧化应激[J].中国神经医学杂志.2004,20(4):327-331.
[3] Schapira AH,Gu M,Taanman JW,et a1.Mitochondria in the etiology and pathogen- nesis of Parkinson’s disease[J].Ann Neurol,1998, 44(Suppl 1):s89-s98.
[4] Hashimoto M,Hsu L J,Xia Y,et a1.Oxidative stress induces Amyloid-like aggregate formation of NACP/alpha-synuclein in vitro[J].Neuroreport. 1999,10:717-721.
[5] 徐叔云.药理实验方法学[M].第 2 版.北京:人民卫生出版社.1991,440-446.
[6] 周宜灿,陈晓眷,朱元贵,等.N-乙酰半胱氨酸防止帕金森病鼠黑质神经元凋亡[J].中国神经科学杂志.2003 Nov,19(6):384-388.
[7] 赵磊,蒲小平.类叶升麻苷对MPTP所致帕金森病小鼠模型的神经保护作用[J].中国药理学通报.2007,23(1) :42-46.
[8] 郭续文.利福平保护MPTP所致PD小鼠中脑多巴胺能神经元凋亡的实验研究[J].中华神经医学杂志.2006,6(5):546-549.
[9] Often D,Beart PM,Cheung NS,et a1.Transgenic mice expressing human bel-2 in their neurons are resistant to 6-hydroxydopamine and 1-methyl-4-phenyl-1,2,3,6- tetrahydropyridine neurotoxicity[J].Proc Natl Acad Sci USA.1998,95(10):5789- 5789.
[10] 丁正同,任惠民,蒋雨平等.百草枯对小鼠黑质纹状体多巴胺能系统的影响[J].复旦学报.2001,Jan 28(1):28-31.
[11] Holthoff VA,Vieregge P,Kessler J,et al.Discordant twins with Parkinson’disease:positron emission tomography and early signs of impaired cognitive circuits[J]. Ann Neurol.1994,36(2):176-182.
[12] Yoshino H,Hattori Y,ImaiH,et al.Sparteine oxidation by hepatic Cytochrome P-450 in patients with Parkinson’s disease[J].RinshoShinkeigaku.1993,33(3): 261-265.
[13] Iacopino AM,Christakos S.Specific reduction of calcium-binding protein (28- kilodaltm calbindin-D)gene expression in aging and neurodegenerative diseases [J].Proc Natl Acad Sci USA.1990,87(11):4078-4082.
[14] Gerlach M,Riederer P,Przuntek H,et a1.MPTP mechanisms of neurotoxicity and their implications for Parkinson’s disease[J].Eur J Pharmacol.1991,208(4):273- 286.
[15] 张如意,张兰,叶翠飞等.MPTP 对小鼠行为学及脑纹状体多巴胺含量的影响[J].中国行为医学科学杂志.2001,10(3):164-166.
[16] Cohen G,Fahn S.The oxidant stress hypothesis in Parkinson’s disease: Evidence supporting it[J].Ann Neurol.1992,32:804-8l2.
[17] Zhang P,Land W,Lee S,et alElectron tomography of degenerating neurons in micewith abnormal regulation of iron metabolism[J].J Struct Biol.2005, 150(2): 144- 153.
[18] Jenner P.Oxidative stress in Parkinson’s disease[J].Ann Neurol.2003, 53(suppl3): S26-36.
[19] Bilsland J,Roy S,Xanthoudakis S,et a1.CASPASE inhibitors attenuate 1-methyl-4- phenylpyridinium toxicity in primary cultures of mesencephatic dopaminergic neurons[J].Neurosci.2002,22(7):2637-2649.
[20] Xu Z,Cawthon D,McCastlain K A,et a1.Selective alterations of gene expression in mice induced by MPTP[J].Synapse.2005,55:45-51.
[21] Kingsbury AE,Mardsen CD,Foster OJ.DNA fragmentation in human substantia nigra:apeptosis or perimortem effect?[J].Mov Disord.1998,13(6):877-884.
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