迷走神经和DVC在胆碱能抗炎通路中的作用及机制的研究

发布时间：2018-06-23 17:15

本文选题：炎症反应 + 辣椒素　；参考：《山东师范大学》2011年硕士论文

【摘要】：胆碱能抗炎通路(cholinergic anti-inflammatory pathway,CAP)是指当致病细菌入侵时,体内的迷走神经及其递质乙酰胆碱(Acetylcholine,Ach)与免疫系统相互作用参与抗炎。已有的研究表明,迷走传出神经及其递质Ach参与了胆碱能抗炎通路的调控过程。黄健等通过先夹伤迷走神经干中枢端,注射内毒素后,发现迷走神经传出纤维放电频率增加。Borovikova等直接电刺激内毒素血症大鼠的迷走神经传出纤维,可抑制肝脏和心脏肿瘤坏死因子-α(TNF-α)的生成,降低血清TNF-α的含量。Thomas等研究显示,电刺激迷走神经能够抑制缺血再灌注损伤所导致的全血TNF-α升高,缓解缺血再灌注引起的休克。石德光、胡森等切断迷走神经则使心肌的TNF-α水平显著升高,炎性损伤加重;电刺激切断的迷走神经远端能使内毒素(LPS)血症大鼠肺组织中TNF-α含量显著降低,炎性病理改变减轻。以上主要通过夹伤、电刺激或切断迷走神经的方法来研究迷走传出神经对炎症反应的影响,但在迷走传出神经正常的情况下,其在炎症反应中放电频率变化,无相关报道。迷走传出神经及其递质Ach参与了抗炎反应,表明炎症反应中迷走传出神经位于迷走神经背核(DMV)内的节前神经元被激活,那么,炎症信息是如何传递至DMV的?通常认为周围免疫信息可通过两种途径传达到脑,一是血液中的细胞因子通过缺乏血脑屏障的最后区(AP)入脑,由AP直接传递至DMV,或由AP经孤束核(NTS)间接传递至DMV。二为细胞因子通过迷走传入神经将炎症信息传至NTS,由NTS传到DMV。胆碱能抗炎通路中炎症信号主要是通过缺乏血脑屏障的AP入脑还是通过迷走传入神经或两者兼之,还不明确。“延髓内脏带”(MVZ)含有儿茶酚胺(CA)类等多种神经活性物质。一系列研究表明MVZ内CA能神经元参与了心血管和胃肠道功能的调节,如脑出血急性期延髓内脏带内儿茶酚胺能神经元有Fos蛋白表达;大鼠在束缚浸水应激状态下DVC内Fos/TH表达增加。而内毒素炎症时,脑干NTS、DMV和AP内的儿茶酚胺能神经元是否参加了炎症信息的传递、分析和整合,有关该方面的研究至今尚未见报道。因此,本文拟采用辣椒素去神经化的方法观察内毒素炎症时对迷走神经传出纤维放电、血清TNF-α水平及DVC内c-Fos+TH表达的影响,以探讨迷走神经和DVC在胆碱能抗炎通路中的作用及其机制。本研究分为三部分: 第一部分,观察内毒素(LPS)炎症时大鼠迷走神经放电、血压、血清TNF-α水平以及DVC内c-Fos表达情况。实验分两组:LPS组和对照(CON)组,LPS组静脉注射LPS;CON组静脉注射等容量的生理盐水。结果与结论:在LPS组,与LPS注射前相比迷走神经放电频率显著增加,血压降低但不显著;LPS组与CON组相比,神经放电频率和血清TNF-α水平显著升高,脑干NTS、DMV和AP的c-Fos表达均显著增强。提示迷走神经、AP和NTS参与了炎症反应。第二部分,辣椒素去神经化后再给予LPS刺激致炎,观察大鼠迷走神经传出纤维放电、血压、血清TNF-α水平的变化以及脑干DVC内c-fos表达情况。实验分为两组:辣椒素+内毒素(CAS+LPS)组和LPS组,CAS+LPS组先以辣椒素去神经化后再静脉注射LPS刺激致炎;LPS组直接静脉注射等容量的LPS。结果与结论:在CAS+LPS组,与LPS注射前相比迷走神经传出纤维放电频率显著增加,血压升高但不明显;CAS+LPS组与LPS组相比,血清TNF-α浓度显著升高,脑干NTS、DMV和AP内c-fos表达均显著减弱。提示内毒素炎症时,迷走传出神经纤维兴奋增多,参入了抗炎症作用。外周炎症信息主要通过迷走传入神经传递至NTS,由NTS传到DMN,部分炎症信号可通过AP传至DMN。第三部分,利用Fos蛋白与酪氨酸羟化酶(TH)免疫双标的方法,观察LPS炎症时对脑干NTS、DMV和AP内CA能神经元活动的影响。实验分为两组:LPS组和CON组,LPS组静脉注射LPS;CON组静脉注射生理盐水。结果与结论:LPS组与CON组相比,LPS炎症时,NTS、DMV和AP内TH阳性神经元和TH+Fos双标阳性神经元数目明显增多,表明儿茶酚胺能神经元参与了炎症反应。结果提示内毒素炎症时,脑干NTS、DMV和AP内的儿茶酚胺能神经元可能参与了胆碱能抗炎症通路炎症信息的传递和炎症反应的调节过程。
[Abstract]:The cholinergic anti-inflammatory pathway (cholinergic anti-inflammatory pathway, CAP) refers to the interaction of the vagus nerve and its neurotransmitter acetylcholine (Acetylcholine, Ach) and the immune system in the body when the pathogenic bacteria invade. The research has shown that the vagal efferent nerve and its transmitter Ach participate in the regulation of the cholinergic anti-inflammatory pathway. After injecting endotoxin to the central end of the vagus nerve and injecting endotoxin, Huang Jian found that the frequency of the discharge of the vagus nerve was increased by.Borovikova and other direct electrical stimulation of the vagus nerve efferent fibers in the rats with endotoxemia, which could inhibit the formation of the liver and cardiac tumor necrosis factor - alpha (TNF- alpha) and reduce the content of TNF- alpha in serum.Thomas. The electrical stimulation of the vagus nerve can inhibit the increase of TNF- alpha in the whole blood caused by ischemia-reperfusion injury and relieve the shock caused by ischemia-reperfusion. Shi Deguang, Honson and other vagus nerves make the TNF- alpha level of the myocardium significantly increased and the inflammatory injury aggravated; the electric stimulation of the vagus nerve can make the lungs of the endotoxemia (LPS) rat lung. The content of TNF- alpha in the tissue decreased significantly and the inflammatory pathological changes were alleviated. The effects of the vagal efferent nerve on the inflammatory response were studied mainly by the method of clamp injury, electrical stimulation or vagotomy, but the frequency of discharge in the inflammatory response was changed without a related report.
The vagal efferent nerve and its transmitter Ach participate in the anti-inflammatory response, indicating that the preganglionic neurons in the dorsal nucleus of the vagus nerve are activated in the inflammatory response, then how does the inflammatory information transfer to DMV? It is generally believed that the peripheral immune information can be communicated to the brain in two ways, one is that the cytokines in the blood pass through the cell. The final region of the blood-brain barrier (AP) is transferred into the brain, from AP to DMV, or by AP via the nucleus of the solitary tract (NTS) to DMV. 2. The inflammatory information passes through the vagal afferent nerve to NTS, and from NTS to the DMV. cholinergic anti-inflammatory pathway, the inflammatory signal is mainly through the AP into the brain that lacks the blood-brain barrier or through the vagus. It is not clear that the "medullary visceral zone" (MVZ) contains a variety of neuroactive substances such as catecholamine (CA). A series of studies have shown that the CA neurons in MVZ are involved in the regulation of cardiovascular and gastrointestinal function, such as the expression of the Fos protein in the catechinamamine neurons in the visceral visceral zone of the cerebral hemorrhage, and the rats are bound and impregnated. In the state of water stress, the expression of Fos/TH in DVC increases. And when endotoxin inflammation, whether the catecholamine neurons in NTS, DMV and AP in the brain stem participate in the transmission of inflammatory information, analysis and integration, the study on this aspect has not yet been reported. Therefore, this paper intends to use capsaicin de neurochemical method to observe the endotoxin inflammation in the vagus The effects of nerve efferent fiber discharge, serum TNF- alpha level and the expression of c-Fos+TH in DVC are discussed in order to explore the role and mechanism of vagus and DVC in the cholinergic anti-inflammatory pathway.
This study is divided into three parts:
The first part was to observe the discharges of vagus nerve, blood pressure, serum TNF- alpha and c-Fos expression in DVC during LPS inflammation. The experiment was divided into two groups: LPS group and control group (CON), LPS group intravenous LPS, and CON group intravenous injection of normal saline. Results and conclusion: in LPS group, the frequency of vagus nerve discharge was compared with LPS injection. In group LPS, the frequency of nerve discharge and the level of TNF- alpha in serum were significantly higher than that in group CON, and the expression of c-Fos in NTS, DMV and AP in the brain stem was significantly enhanced. The vagus nerve, AP and NTS were involved in the inflammatory response.
The second part, after the capsaicin was denervated and then stimulated by LPS stimulation, observed the changes in the efferent fibers of the vagus nerve, the changes of blood pressure, the level of serum TNF- A and the expression of c-fos in the brain stem DVC. The experiment was divided into two groups: capsaicin + endotoxin (CAS+LPS) group and LPS group. The CAS+ LPS group first used capsaicin to neurochemical and then intravenously injected LPS stimulation LPS. results and conclusions of direct intravenous injection in LPS group: in group CAS+LPS, the frequency of the efferent fibers in the vagus nerve increased significantly, and the blood pressure increased but was not obvious before LPS injection; the serum TNF- alpha concentration in the CAS+LPS group was significantly higher than that in the LPS group, and the c-fos expression in the brain stem NTS, DMV and AP was significantly weakened. At the time, the excitability of the efferent nerve fibers increased and joined the anti inflammatory effect. The peripheral inflammatory information was transmitted to NTS mainly through the vagal afferent nerve, transmitted from NTS to DMN, and some of the inflammatory signals could be transmitted to DMN. through AP.
In the third part, the effects of Fos protein and tyrosine hydroxylase (TH) immunization were used to observe the effects of LPS inflammation on the activity of CA neurons in the brain stem NTS, DMV and AP. The experiments were divided into groups LPS and CON, LPS group intravenous LPS, CON group intravenous injection of saline. The number of TH positive neurons and TH+Fos double labeled neurons increased significantly, indicating that catecholamine neurons were involved in the inflammatory response. The results suggested that the catecholamine neurons in the brain stem NTS, DMV and AP may participate in the regulation of inflammatory response and inflammatory response in the cholinergic anti-inflammatory pathway.
【学位授予单位】：山东师范大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

【引证文献】