中西医结合临床专业论文03750.doc
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1、Shaanxi University of Chinese Medicine硕士学位论文疏筋解毒汤干预PD大鼠黑质内质网应激的实验研究中文摘要目的:探讨疏筋解毒汤对帕金森病(PD)模型大鼠脑内黑质多巴胺能神经元内质网应激信号传导蛋白基因表达的调节作用。方法:采用向大鼠黑质内定位注射6-OHDA法制备PD大鼠模型。用正常SD大鼠作为空白对照组,并将造模成功的大鼠分为模型组、西药治疗组、疏筋解毒汤小剂量治疗组、疏筋解毒汤中剂量治疗组、疏筋解毒汤大剂量治疗组,共6个组。利用逆转录聚合酶链反应(RT-PCR)法对各实验组大鼠黑质的CHOPmRNA、XBP-1mRNA表达进行分析,检测各个指标在各组中的
2、表达情况。结果:1、CHOPmRNA表达:中药组CHOPmRNA的表达从14d到60d的用药过程中,随时间的延长,CHOPmRNA的表达呈渐降趋势。且在14d、21d、30d、45d、60d 5个时间点均低于西药治疗组和模型对照组,有统计学意义(P0.01)。各组CHOPmRNA的表达在各个时间点均高于空白组。2、XBP-1mRNA表达:中药组XBP-1mRNA表达从14d到60d的用药过程中,随时间的延长,XBP-1mRNA表达呈渐升趋势。且在14d、21d、30d、45d、60d 5个时间点均高于西药治疗组、模型对照组,有统计学意义(P0.01)。结论:1、疏筋解毒汤能明显抑制内质网过度应
3、激时促凋亡因子CHOPmRNA的表达,从而对多巴胺能神经元凋亡有阻抑作用。2、疏筋解毒汤能明显促进未折叠蛋白反应信号传导通路的分子伴侣XBP-1mRNA的表达,从而保护多巴胺能神经元。3、疏筋解毒汤可以通过干预内质网应激保护帕金森大鼠黑质多巴胺能神经元。关键词:疏筋解毒汤;内质网应激;CHOP; XBP-1The experimental research of the endoplasmic reticulum stress in the rats substantia nigra intervened by shujinjiedu TangAbstractPurpose:Probing t
4、he adjusting effect of shujinjiedu Tang to the gene expression of the protein in the conduction of stress signal of substantia nigra dopaminergic neurons endoplasmic reticulum in the model rats infected by parkinsons disease.Methods:6-OHDA is injected into the substantia nigra of rats to make the mo
5、del rats. The normal SD rats are used as the blank control group. Also, the rats that are successfully modeled are divided into six groups, such as the modeling group, the western medicine treatment group, the little, medium and large dose treatment groups with shujinjiedu Tang. RT-PCR method is use
6、d to analyze the expressions of CHOPmRNA, XBP-1mRNA in the substantia nigra of each experimental group, and the expression conditions of every indicator is detected in each group.Results:1. The expression of CHOPmRNA: between the 14th day and the 60th day after the shujinjiedu Tang is used, the expr
7、ession of CHOPmRNA shows a descending tendency with the increasing of the time. Moreover, at the following five moment, such as 14th day, 21st day, 30th day, 45th day, 60th day, the expression of CHOPmRNA is lower than that of the western medicine treatment group and the modeling group, which has ce
8、rtain statistical significance. And the expression of CHOPmRNA in blank control group is lower than that of the other groups at the above five moments.2. The expression of XBP-1mRNA: between the 14th day and the 60th day after the shujinjiedu Tang is used, the expression of XBP-1mRNA shows a increas
9、ing tendency with the increasing of the time. Moreover, at the following five moment, such as 14th day, 21st day, 30th day, 45th day, 60th day, the expression of XBP-1mRNA is higher than that of the western medicine treatment group, the modeling group and the blank control group, which has certain s
10、tatistical significance. Conclusions:1. The expression of the pro-apoptotic factor CHOPmRNA can obviously been constrained by shujinjiedu Tang when the endoplasmic reticulum is over stressed. Therefore, the shujinjiedu Tang have some constraining effect to the dopaminergic neurons apoptosis.2. The e
11、xpression of the molecular chaperones XBP-1mRNA can obviously been promoted by shujinjiedu Tang in the conduction path of the unfolded proteins response signal. Therefore, the shujinjiedu Tang can protect the neurons.3. The dopaminergic neurons in the substantia nigra of the rats infected by the par
12、kinsons disease can be protected by shujinjiedu Tang via interfering its stressing of the endoplasmic reticulum.Key words:shujinjiedu Tang; stressing of the endoplasmic reticulum; CHOP; XBP-1目 录引言.1研究背景.31 西医对帕金森病的认识.31.1 PD的临床表现.31.2 PD的病理变化.31.3 PD的病因及发病机制. .31.3.1 遗传因素. .31.3.2 环境因素. .41.3.3 神经系统
13、老化. 41.3.4 氧化应激及自由及损伤. .41.3.5 兴奋性毒性作用.41.3.6 细胞凋亡.51.3.7 免疫功能变化.51.3.8 内质网应激.51.4 PD的现代治疗方法.71.4.1 药物治疗.71.4.2 细胞移植治疗.81.4.3 外科手术治疗.82 中医对帕金森病的认识.82.1 历代医家对颤证的描述.82.2 颤证的病因病机.92.3 颤证的辨证治疗.102.3.1 用针刺、灸法调理经络治疗.102.3.2 中药辨证施治.103 疏筋解毒汤药理研究.11实验研究.141 实验材料.141.1 实验试剂.141.2 实验仪器. 141.3 实验设计和技术路线.152 PD
14、模型大鼠的制备.172.1 实验动物分组.172.2 实验原理.172.3 模型制备.172.4 灌胃、取组织.183 RT-PCR分析CHOP、XBP-1的表达.193.1 主要试剂配制.193.2 引物序列设计.203.3 实验步骤.203.3.1 总RNA的提取.203.3.2 总RNA逆转录为cDNA.213.3.3 PCR扩增.213.3.4 电泳.223.4 实验结果.233.4.1 分析方法.233.4.2 疏筋解毒方对PD大鼠黑质CHOPmRNA表达的影响.233.4.3 疏筋解毒方对PD大鼠黑质XBP-ImRNA表达的影响.263.5 结论.303.6 讨论.31参考文献.3
15、2致谢.36疏筋解毒汤干预PD大鼠黑质内质网应激的实验研究引 言帕金森病(Parkinson,s Disease,PD)是一种常见于中老年的神经系统变性疾病,临床上以静止性震颤、运动迟缓、肌强直和姿势步态障碍为主要特征1。此外病人还可出现慌张步态、面具脸、小字征、吞咽因难、自主神经功能障碍和痴呆等表现。1817年英国医生James Parkinson首先描述了PD的一系列特殊症候群。本病目前尚无彻底治愈的方法,是目前医学还未能攻克的顽疾。据流行病学调查发现,其发病率有逐年增高并向高龄移动的趋势,我国帕金森病患病率约为160/10万2,在55岁以上的人群中患病率达到1,由此推算我国的患病人数约为
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