苦参碱对阿尔茨海默病模型大鼠海马突触超微结构的影响
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摘要
目的:探讨苦参碱对阿尔茨海默病模型大鼠海马突触超微结构的影响。方法:将Morris水迷宫试验达标的18只雄性SD大鼠,随机分为正常对照组、模型组和苦参碱组(15 mg·kg~(-1)),每组6只。模型组和苦参碱组大鼠双侧海马注射凝聚态Aβ_(1-40),正常对照组分别给予等体积药物或生理盐水灌胃,持续28天。采用流式细胞仪检测各组实验动物海马神经细胞凋亡率的变化,使用TUNEL染色检测海马神经细胞凋亡数,免疫组织化学法检测各组大鼠海马神经细胞细胞色素C变化,在透射电镜下观察海马突触超微结构的改变。结果:流式细胞仪检测及TUNEL染色显示,模型组大鼠海马阳性细胞数明显高于对照组(P <0.01),苦参碱组海马阳性细胞数较模型组明显减少(P <0.01);用药组大鼠海马细胞色素C阳性产物数和平均光密度均较模型组明显减少(P <0.01);透射电镜下显示,对照组海马神经细胞结构、线粒体膜、神经轴突和神经突触结构完整;模型组神经细胞结构、线粒体膜、神经轴突和神经突触结构破坏;苦参碱组大鼠海马线粒体和神经突触基本正常。结论:苦参碱灌胃可减轻大鼠海马超微结构的破坏,减少海马神经细胞凋亡。
Objective: To investigate the effect of Matrine on ultrastructure of hippocampal synapse in Alzheimer's disease model rats. Methods: 18 male SD rats were randomly divided into normal control group, model group, matrine group(15 mg·Kg~(-1)) in Morris water maze test,in which there is 6 rats in each group. The rats in the model group, matrine group were injected with Aβ_(1-40) in the bilateral hippocampus. The rats in the normal control group were given the same volume or normal saline for 28 days. Morris water maze was used to test behavioral indicators, and observed the learning and memory abilities of rats, and observed ultrastructural changes in hippocampal synapses under transmission electron microscope. By flow cytometry, we observed the changes in the rate of neuronal apoptosis in hippocampus of each group.Apoptosis of hippocampal neurons was detected by TUNEL staining. Changes of cytochrome C in rat hippocampal nerve cells were detected by immunohistochemistry. Results: Display the transmission electron microscope, the control group of nerve cells in hippocampus, mitochondrial structure the membrane, axons and synaptic structural integrity; the structure of nerve cells, mitochondrial membrane, axons and synaptic structure damaged in model of group. But in matrine group,the hippocampal mitochondria and synaptic of rat are normal; Flow cytometry and the TUNEL staining showed that the number of positive cells in hippocampus of rats in the model group was significantly higher than that in the control group(P < 0.01), in matrine group the number of positive cells decreased compared with that in model group(P < 0.01). The number of positive products and the hippocampal cytochrome C in the treated group were significantly lower than those in the model group. Conclusion: Matrine can reduce the ultrastructural damage of hippocampus, and reduce the apoptosis of hippocampal neurons.
Objective: To investigate the effect of Matrine on ultrastructure of hippocampal synapse in Alzheimer's disease model rats. Methods: 18 male SD rats were randomly divided into normal control group, model group, matrine group(15 mg·Kg~(-1)) in Morris water maze test,in which there is 6 rats in each group. The rats in the model group, matrine group were injected with Aβ_(1-40) in the bilateral hippocampus. The rats in the normal control group were given the same volume or normal saline for 28 days. Morris water maze was used to test behavioral indicators, and observed the learning and memory abilities of rats, and observed ultrastructural changes in hippocampal synapses under transmission electron microscope. By flow cytometry, we observed the changes in the rate of neuronal apoptosis in hippocampus of each group.Apoptosis of hippocampal neurons was detected by TUNEL staining. Changes of cytochrome C in rat hippocampal nerve cells were detected by immunohistochemistry. Results: Display the transmission electron microscope, the control group of nerve cells in hippocampus, mitochondrial structure the membrane, axons and synaptic structural integrity; the structure of nerve cells, mitochondrial membrane, axons and synaptic structure damaged in model of group. But in matrine group,the hippocampal mitochondria and synaptic of rat are normal; Flow cytometry and the TUNEL staining showed that the number of positive cells in hippocampus of rats in the model group was significantly higher than that in the control group(P < 0.01), in matrine group the number of positive cells decreased compared with that in model group(P < 0.01). The number of positive products and the hippocampal cytochrome C in the treated group were significantly lower than those in the model group. Conclusion: Matrine can reduce the ultrastructural damage of hippocampus, and reduce the apoptosis of hippocampal neurons.
引文
1 Alzheimer's Association.2018 Alzheimer's disease facts and figures.Alzheimers Dement,2018,14(3):367-429.
2王延江,卜先乐.阿尔茨海默病的防治研究进展.第三军医大学学报,2014,36(21):2153-2157.
3黄涛波,吕诚,胡小令,等.雷公藤内酯醇对AD模型大鼠海马神经细胞凋亡的影响.中国老年学,2010,30(19):2766-2770.
4 Goldman J G,Holden S.Treatment of psychosis and dementia in Parkin-son's disease.Curr Treat Options Neurol,2014,16(3):281-285.
5桂婷.雷公藤内酯醇对阿尔茨海默病模型大鼠海马突触后致密物蛋白N-甲基-D-天门冬氨酸受体亚基2B和突触后致密物质95的影响.南昌:南昌大学硕士研究生学位论文,2010.
6王婷.何首乌二苯乙烯苷对缺血性脑损伤和学习记忆的影响及其机制.武汉:华中科技大学博士研究生学位论文,2009.
7和欢,尚新悦,宋少江.苦参碱及其制剂的研究进展.世界科学技术-中医药现代化,2016,18(7):1099-1107.
8张翅,马悦,高慧敏,等.苦参化学成分研究进展.中国实验方剂学杂志,2014,20(4):205-214.
9杨新阶,王婧筱,王琳,等.中药标准品对人肺癌耐药细胞A549/DDP的逆转效果研究.世界科学技术-中医药现代化,2018,20(4):521-526.
10林文,张俊平,胡振林,等.苦参碱对细菌脂多糖诱导大鼠枯否细胞释放肿瘤坏死因子及白细胞介素-6的影响.药学学报,1997,32(2):93-96.
11王俊学,王国俊.苦参碱及氧化苦参碱的药理作用及临床应用.肝脏,2000,5(2):116-117.
12姚遥,孙娜,郑萍,等.苦参碱对阿尔茨海默症小鼠学习记忆功能和脑内氧化应激的影响.宁夏医科大学学报,2016,38(9):985-988.
13李娟,姚遥,韩怀钦,等.苦参碱对LPS诱导的阿尔茨海默病小鼠模型学习记忆功能和脑内神经炎症的影响.中国实验方剂学杂志,2018,24(24):1-6.
14封敏,卢圣锋,张承舜,等.国内大鼠Morris水迷宫实验现状与分析.辽宁中医杂志,2011,28(11):2170-2173.
15武海霞,吴志刚,刘红彬,等.Morris水迷宫实验在空间学习记忆研究中的应用.神经药理学报,2014,5(4):30-35.
16 Paxinos G,Watson C.The rat brain in stereotaxic coordiantes.Elsevier:Academie Press,1986:2.
17李宜培,彭蕤蕤,柴高尚,等.促红细胞生成素对阿尔茨海默病模型大鼠记忆能力及其超微结构的影响.中国老年学杂志,2015,35(3):1620-1622.
18微结构的影响.解剖学杂志,2010,33(6):716-719.
19黄涛波,吕诚,胡小令,等.雷公藤内酯醇对AD模型大鼠海马神经细胞凋亡的影响.中国老年学杂志,2010,30(10):2766-2770.
20 Krause D L,Müller N.Microglial activation and neuroinflammation in alzheimer's disease:a critical examination of recent history.Front Aging Neurosci,2010,2:22.
21 Calissano P,Matrone C,Amadorov G.Apoptosis and in vitro Alzheimerdisease neuronal models.Commun Integr Biol,2009,2(2):163-169.
22王瑛,张海音.阿尔茨海默病的发病机制研究进展.中国神经免疫学和神经病学杂志,2002,9(1):42-48.
23朱晓蕾.氧化应激在阿尔茨海默病发病机制中的作用及其靶点药物的开发.南京:南京中医药大学博士研究生学位论文,2012.
24 Griffin W S,SHENG J G,Royston M C,et al.Glial-neuronal interactions in Alzheimer's disease:the potential role of a‘cytokine cycle’in disease progression.Brain Pathol,1998,8(1):65-72.
2王延江,卜先乐.阿尔茨海默病的防治研究进展.第三军医大学学报,2014,36(21):2153-2157.
3黄涛波,吕诚,胡小令,等.雷公藤内酯醇对AD模型大鼠海马神经细胞凋亡的影响.中国老年学,2010,30(19):2766-2770.
4 Goldman J G,Holden S.Treatment of psychosis and dementia in Parkin-son's disease.Curr Treat Options Neurol,2014,16(3):281-285.
5桂婷.雷公藤内酯醇对阿尔茨海默病模型大鼠海马突触后致密物蛋白N-甲基-D-天门冬氨酸受体亚基2B和突触后致密物质95的影响.南昌:南昌大学硕士研究生学位论文,2010.
6王婷.何首乌二苯乙烯苷对缺血性脑损伤和学习记忆的影响及其机制.武汉:华中科技大学博士研究生学位论文,2009.
7和欢,尚新悦,宋少江.苦参碱及其制剂的研究进展.世界科学技术-中医药现代化,2016,18(7):1099-1107.
8张翅,马悦,高慧敏,等.苦参化学成分研究进展.中国实验方剂学杂志,2014,20(4):205-214.
9杨新阶,王婧筱,王琳,等.中药标准品对人肺癌耐药细胞A549/DDP的逆转效果研究.世界科学技术-中医药现代化,2018,20(4):521-526.
10林文,张俊平,胡振林,等.苦参碱对细菌脂多糖诱导大鼠枯否细胞释放肿瘤坏死因子及白细胞介素-6的影响.药学学报,1997,32(2):93-96.
11王俊学,王国俊.苦参碱及氧化苦参碱的药理作用及临床应用.肝脏,2000,5(2):116-117.
12姚遥,孙娜,郑萍,等.苦参碱对阿尔茨海默症小鼠学习记忆功能和脑内氧化应激的影响.宁夏医科大学学报,2016,38(9):985-988.
13李娟,姚遥,韩怀钦,等.苦参碱对LPS诱导的阿尔茨海默病小鼠模型学习记忆功能和脑内神经炎症的影响.中国实验方剂学杂志,2018,24(24):1-6.
14封敏,卢圣锋,张承舜,等.国内大鼠Morris水迷宫实验现状与分析.辽宁中医杂志,2011,28(11):2170-2173.
15武海霞,吴志刚,刘红彬,等.Morris水迷宫实验在空间学习记忆研究中的应用.神经药理学报,2014,5(4):30-35.
16 Paxinos G,Watson C.The rat brain in stereotaxic coordiantes.Elsevier:Academie Press,1986:2.
17李宜培,彭蕤蕤,柴高尚,等.促红细胞生成素对阿尔茨海默病模型大鼠记忆能力及其超微结构的影响.中国老年学杂志,2015,35(3):1620-1622.
18微结构的影响.解剖学杂志,2010,33(6):716-719.
19黄涛波,吕诚,胡小令,等.雷公藤内酯醇对AD模型大鼠海马神经细胞凋亡的影响.中国老年学杂志,2010,30(10):2766-2770.
20 Krause D L,Müller N.Microglial activation and neuroinflammation in alzheimer's disease:a critical examination of recent history.Front Aging Neurosci,2010,2:22.
21 Calissano P,Matrone C,Amadorov G.Apoptosis and in vitro Alzheimerdisease neuronal models.Commun Integr Biol,2009,2(2):163-169.
22王瑛,张海音.阿尔茨海默病的发病机制研究进展.中国神经免疫学和神经病学杂志,2002,9(1):42-48.
23朱晓蕾.氧化应激在阿尔茨海默病发病机制中的作用及其靶点药物的开发.南京:南京中医药大学博士研究生学位论文,2012.
24 Griffin W S,SHENG J G,Royston M C,et al.Glial-neuronal interactions in Alzheimer's disease:the potential role of a‘cytokine cycle’in disease progression.Brain Pathol,1998,8(1):65-72.
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