【作者】 吴北峰；

【导师】 高维滨；

【作者基本信息】 黑龙江中医药大学 ， 针灸推拿学， 2008， 博士

【摘要】 本论文是在以往对电项针临床应用和研究的基础上,对电项针治疗失眠的作用作了系统全面的总结、归纳后,通过睡眠剥夺大鼠动物模型,观察其脑干网状结构中单胺类神经递质、脑肠肽、NO和SOD含量的变化,从分子水平对电项针治疗失眠及其氧化应激反应进行深入研究。目的:1通过电项针干预睡眠剥夺大鼠的实验研究探讨电项针对失眠及其诱发氧化应激反应影响机制。方法:将大鼠随机分为空白组、模型组、电项针组、甜梦胶囊组、黄芪注射液组,除空白组外,其余组通过腹腔注射PCPA复制睡眠剥夺大鼠模型。1观察对比睡眠剥夺5天后各组大鼠体重变化。2通过电镜观察大鼠神经细胞的变化情况。3通过荧光分光光度计观察大鼠脑中5-HT、5-HIAA、NE含量的变化。4通过光栅分光光度计观察脑干中SOD及NO含量的变化。5通过免疫组化染色法观察CCK8、GABA在睡眠剥夺大鼠脑干组织中阳性表达情况。结果:1模型组大鼠脑干网状结构中5-HT、5-HIAA、GABA、CCK8含量降低,NO、SOD、NE含量升高,与空白组对比有显著差异(P＜0.01),理论上造模成功。2睡眠剥夺5天后,模型组大鼠体重减轻,与空白组比较有明显差异(P＜0.01),电项针组大鼠体重无明显变化,与空白组无明显差异(P＞0.05)。3电镜下观察可见,睡眠剥夺后,模型组与甜梦胶囊组大鼠脑干细胞膜边界不清晰,细胞浑浊,细胞内有空泡变性,线粒体空泡变性,肿胀明显粗面内质网含量减少,突触融合。而电项针组未见明显变化。4荧光分光光度计法证实,睡眠剥夺后,电项针可以提高脑干网状组织内5-HT、5-HIAA的含量,降低NE的含量,与模型组比较有显著差异(P＜0.01),甜梦胶囊组NE的含量降低,同模型组相比有显著性差异(P＜0.01),与电项针组比较无显著性差异(P＞0.05)。甜梦胶囊组5-HT、5-HIAA的含量降低,同模型组相比无显著性差异(P＞0.05)。与电项针比较有显著性差异(P＜0.05)。5光栅分光光度计法证实,睡眠剥夺后,电项针组和黄芪注射液组大鼠脑干网状组织内NO、SOD的含量降低,与模型组比较均有显著差异(P＜0.01)。两组比较无显著性差异(P＞0.05)。6免疫组织化学染色法证实,睡眠剥夺后,电项针组和甜梦胶囊组大鼠脑干网状组织内CCK8的含量升高,与模型组比较有显著差异(P＜0.01),两组比较无显著性差异(P＞0.05)。电项针组和甜梦胶囊组大鼠脑干网状组织内GABA的含量升高,与模型组比较,电项针组有显著差异(P＜0.01),甜梦胶囊组无明显差异(P＞0.05),两组比较有显著性差异(P＜0.05)。结论:1睡眠剥夺可以破坏大鼠脑干中的神经细胞,而电项针对神经细胞具有保护作用,抑制睡眠剥夺对大鼠脑干神经细胞的损害。2电项针可以通过恢复5-HT通路与NE通路之间的相互平衡和制约,进而恢复正常的睡眠一觉醒节律,从而纠正失眠。3电项针可通过增强GABA的抑制作用,从而实现治疗失眠的目的。4电项针可通过恢复CCK8与NO通路之间的平衡减轻睡眠剥夺大鼠的氧化应激反应,减少睡眠剥夺大鼠神经毒性作用。通过增加CCK8减轻大鼠由睡眠剥夺诱发氧化应激引起进食增多,体重减轻的高度能量消耗的营养不良状态。更多还原

【Abstract】 The electro-nape-acupuncture has been widely used in clinical Practice ever Since they were recorded.More and more doctors have used it to cure diseases based on the clinical needs.This dissertation makes a comprehensive and Systematic research on their medical effects on insomnia and then,takes a further study on the effect of electro-nape-acupuncture in curing insomnia.To observe the content of mechanism of monoamine neurotransmitter,brain-gut peptide(BGP),nitric oxide (NO)and the activity of super oxide dismutase(SOD)in rat brain-stem reticular formation after sleep deprivation,making a Research of electro-nape-acupuncture curing insomnia and its oxidative stress at the molecular levelObjective:To discuss the mechanism of the electro-nape-acupuncture on the effect of rats of sleep deprivation and oxidative stress induced by it through the intervention of sleep deprivation experiments on rat.Methods:Rats were randomly divided into control group,model group,the acupuncture group,and Sweet Dream Capsule group,astragalus injection group,In addition to the control group,the other group were duplicated of sleep deprivation rat model by intraperitoneal injection of PCPA1 Compared the change of body weight rats in each group after five days.of sleep deprivation.2 Comparing rat neuronal cell changes,through electron microscopy3 Comparing by fluorescence spectrophotometer the content of 5-HT and 5-HIAA,and NE.4 Confirming the positive expression of CCK8,GABA in the rat brainstem. Reticular structure,by immunohistochemical staining,5 Observing the content of NO and SOD.by grating spectrophotometer methodResults:1 after five days of sleep deprivation,the model rats weight decrease,compared with the control group the difference is significant(P＜0.01),The electro-nape-acupuncture group had no significant change in body weight,compared with the control group there was no significant difference(P＞0.05). 2 Observe by electron microscopy,in the model group and the Sweet Dream Capsule group,the borders of cells in brain stem of rats is not clear,Mitochondria degeneration clearly,the content of rough endoplasmic reticulum decrease,synaptic integrate,the cells of rats in the electro-nape-acupuncture group do not change significantly.3 Confirmed by fluorescence spectrophotometer,The electro-nape-acupuncture can increase the content of 5-HT and 5-HIAA,in the brainstem reticularof of sleep deprivation rats,decrease the levels of NE.compared with the model group the difference is significant(P＜0.01),in the Sweet Dream Capsule group,the content of 5-HT and 5-HIAA increase,compared with the model group the difference is not significant(P＞0.05).compared with the electro-nape-acupuncture group the difference is significant(P＜0.05).the content of NE decrease,compared with the model group the difference is significant(P＜0.01),compared with the electro-nape-acupuncture group,the difference is not significant(P＞0.05)4 Confirmed by immunohistochemical staining,The electro-nape-acupuncture and the Sweet Dream Capsule can increase the positive expression of CCK8 in the rat brainstem,reticular structure,Compared with the model group there were significant differences(P＜0.01),Compared with each other,the difference is not significant(P＞0.05).The electro-nape-acupuncture and the Sweet Dream Capsule can increase the positive expression of GABA in the rat brainstem,reticular structure, Compared with the model group there were significant differences in electro-nape-acupuncture group(P＜0.01),there were no significant differences in the Sweet Dream Capsule group(P＞0.05),Compared with each other,the difference is significant(P＜0.01).5 confirmed by grating spectrophotometer method,The electro-nape-acupuncture and astragalus injection can decrease the content of NO and SOD,in the brainstem reticular of the sleep deprivation rats Compared with the model group there were significant differences(P＜0.01).Compared with each other,the difference is no significant(P＞0.05).Conclusion: 1 Sleep-deprivation can damage nerve cells in the rat brainstem,and the powe of the electro-nape-acupuncture has is to relieve the damage of Neuvel cells and protect it.2 The electro-nape-acupuncture can resumpte 5-HT-NE pathway and the mutual balance between access and constraints,thereby restoring normal rhythm of sleep and awakening3 The electro-nape-acupuncture cures insomnia by enhancing inhibitory effect of GABA in order to achieve improvement in the role of sleep.4 The electro-nape-acupuncture relieve the damage of weight loss,the high energy consumption of the state of malnutrition induced by sleep deprivation and oxidative stress through increasing the content of CCK8.It passed through the resumption of CCK8 and NO pathway and the balance NO and CCK8 to reduce the oxidative stress response,Reduce rats neurotoxicity caused by sleep deprivation.更多还原