论文部分内容阅读
背景:作为一种强效血管和神经活性肽,内皮素1在各种中枢神经系统病理生理情况下表达增加,可能对神经组织产生有害作用。然而,增高的内皮素1有无直接诱导神经元凋亡的作用尚不清楚。目的:观察内皮素1有无直接诱导原代培养神经元凋亡的作用。设计:以细胞为研究对象,完全随机对照实验研究。单位:一所大学医院的神经内科和一所大学的病理学教研室、生命科学技术学院组织与移植免疫实验中心。材料:实验于2001-03/2002-02在暨南大学医学院病理教研室和生命科学和技术学院组织移植与免疫研究所实验室进行。来自于新生大鼠大脑皮质的原代神经元培养,新生大鼠由中山大学医学中心动物研究所提供。干预:原代大脑皮质神经元培养5d后分别加入0.2,20nmol/L内皮素1处理24h,用AnnexinV,Hoechst33258染色半定量测定细胞凋亡。再用流式细胞仪分别定量检测内皮素受体A拮抗剂或内皮素受体B拮抗剂对20nmol/L内皮素1诱导神经元凋亡的效果。对照组加入等量磷酸盐缓冲液。主要观察指标:观察内皮素1直接诱导培养神经元凋亡的作用,以及其通过的ET受体亚型。结果:加入0.2nmol/L内皮素124h后,Annexin-V和Hoechst33258阳性染色细胞率分别为(23.00±9.96)%,(9.82±0.95)%,与对照组相比差异无显著性意义犤(Annexin-V:(13.50±3.35)%;Hoe
BACKGROUND: As a potent vascular and neuroactive peptide, endothelin 1 is expressed in a variety of central nervous system pathophysiological conditions and may have detrimental effects on nervous tissue. However, the role of increased endothelin 1 in the direct induction of neuronal apoptosis is unclear. Objective: To observe the effect of endothelin-1 on the apoptosis of primary cultured neurons. Design: Cell-based research, a completely randomized controlled experimental study. Unit: Department of Neurology, University Hospital and Department of Pathology, College of Life Science and Technology, Institute of Immunology and Immunology. MATERIALS: The experiment was performed in the Laboratory of Pathology and Department of Pathology, School of Medicine, Jinan University from March 2001 to February 2002 and the Institute of Tissue Transplantation and Immunology, College of Life Science and Technology. Primary neurons from neonatal rat cerebral cortex were cultured and neonatal rats were provided by the Institute of Zoology, Sun Yat-sen Medical Center. Intervention: primary cultured cerebral cortex neurons were treated with 0.2, 20 nmol / L endothelin 1 for 5 days, respectively, and then treated with endothelin 1 for 24 h. The apoptosis of the neurons was detected by Annexin V and Hoechst33258 staining. The effects of endothelin receptor A antagonist or endothelin receptor B antagonist on neuron apoptosis induced by ET-1 at a concentration of 20nmol / L were determined by flow cytometry. The control group added the same amount of phosphate buffer. MAIN OUTCOME MEASURES: The effect of endothelin-1 on direct induction of neuronal apoptosis and the ET receptor subtype it passed were observed. Results: The percentage of Annexin-V and Hoechst33258-positive cells was (23.00 ± 9.96)% and (9.82 ± 0.95)% respectively after adding 0.2nmol / L endothelin for 124h, which showed no significant difference compared with the control group -V: (13.50 ± 3.35)%; Hoe