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背景:脂多糖(lipopolysaccharide,LPS)作为一种多克隆免疫激发剂刺激免疫细胞,来模拟免疫激发状态,观察延髓内脏带(medullaryvisceralzone,MVZ)投射至下丘脑室旁核(paraventricularnucleusofhypothalamus,PVN)的儿茶酚胺神经元是否对LPS刺激起反应,为脑功能保护研究提供理论依据。目的:观察MVZ投射至PVN的儿茶酚胺神经元是否对脂多糖刺激起反应,探讨MVZ至PVN的儿茶酚胺能通路在“免疫-脑通讯”中的作用。设计:以实验动物为研究对象,随机对照的研究。单位:一所军医大学的神经外科和神经科学研究所。材料:实验在南方医科大学珠江医院神经外科和解放军解放军第四军医大学全军神经科学研究所完成。10只健康清洁级成年SD大鼠由第四军医大学实验动物中心提供。方法:将WGA-HRP注入大鼠一侧PVN内,存活48h后经腹腔注射免疫刺激剂LPS诱发免疫反应,应用WGA-HRP逆行追踪与抗Fos和抗酪氨酸羟化酶(TH)抗体双重免疫组织化学相结合三重标记方法进行染色。主要观察指标:观察MVZ中WGA-HRP逆标细胞、Fos蛋白和儿茶酚胺能神经元(以TH为标记物)的分布及表达状况。结果:在MVZ内发现7种阳性神经元,即HRP,Fos和TH单标细胞,Fos/HRP,Fos/TH和HRP/TH双标细胞,Fos/HRP/TH三标细胞。Fos/HRP双标记和Fos/HRP/TH三标记细胞分别占总HRP逆
BACKGROUND: Lipopolysaccharide (LPS), as a polyclonal immunostimulator, stimulates immune cells to simulate the immune activation state and to observe the catecholamines that are projected by the medullaryvisceralzone (MVZ) into the paraventricular nucleus of hypothalamus (PVN) Whether neurons respond to LPS stimulation provides a theoretical basis for the study of brain function protection. OBJECTIVE: To investigate whether catecholaminergic neurons projecting to PVN in MVZ respond to lipopolysaccharide stimulation and to explore the role of catecholaminergic pathway in MVZ to PVN in “immune-brain communication”. Design: A study of experimental animals, randomized controlled studies. Unit: Institute of Neurosurgery and Neuroscience, a Military Medical University. MATERIALS: Experiments were performed at Department of Neurosurgery, Zhujiang Hospital, Southern Medical University and PLA Institute of Neuroscience, Fourth Military Medical University of the People’s Liberation Army. Ten healthy and clean adult SD rats were provided by Experimental Animal Center of the Fourth Military Medical University. Methods: The WGA-HRP was injected into the PVN of rats and survived for 48 hours. The immune response was induced by intraperitoneal injection of LPS. The WGA-HRP retrograde tracing was used to detect the anti-Fos and TH tyrosine antibodies Immunohistochemistry combined with triple labeling method for staining. MAIN OUTCOME MEASURES: The distribution and expression of WGA-HRP negative cells, Fos protein and catecholaminergic neurons (labeled with TH) in MVZ were observed. RESULTS: Seven positive neurons were found in MVZ, namely HRP, Fos and TH single cells, Fos / HRP, Fos / TH and HRP / TH double labeled cells and Fos / HRP / TH triplets. Fos / HRP double labeling and Fos / HRP / TH triple labeling cells accounted for the total HRP inverse