论文部分内容阅读
目的:ApoE基因敲除小鼠为目前研究动脉粥样硬化的常用模型之一,其在高脂饲养下可加速形成主动脉粥样硬化斑块。动脉粥样硬化的病灶成分有平滑肌细胞、泡沫细胞、胆固醇结晶、胶原纤维、弹力纤维和蛋白聚糖。Movat染色技术可以在一张切片上直观显示出这几种成分的病理变化,规范和细化的实验操作步骤可以保证染色结果的一致性,为动脉粥样硬化的深入研究提供清晰可靠的实验依据,从而更客观的评估实验结果。方法:Apo E基因敲除小鼠,雄性70只随机分为8组:Apo E空白组,阳性对照药组,治疗药物A高、中、低剂量组,治疗药物B高、中、低剂量组,以及6只C57小鼠对照组。高脂饲养26周后,采血后处死。取心脏,石蜡包埋后进行定位切片。分批进行染色,染色过程中摸索和细化了染液配置、染色温度、染色时间等条件,规范实验操作。成片后进行组织病理学观察,并对胶原纤维、弹力纤维、蛋白聚糖、泡沫细胞等进行定性和定量分析。结果:通过试验建立了细化、规范化的Movat染色操作流程。光镜下观察染色结果可见各结构成分着色分明:胶原纤维和弹力纤维分别为黄色和黑色,蛋白聚糖呈绿色,泡沫细胞呈淡紫色,心肌平滑肌呈红色。主动脉粥样硬化斑块病灶部位结构清晰,病理表现为血管平滑肌增生,胶原纤维、平滑肌细胞和蛋白聚糖形成纤维帽,纤维帽下泡沫细胞形成并向下延伸,脂质大量沉积,弹力纤维断裂。与空白组和阳性对照组相比,治疗药物A和B均对病症的严重程度和病程进展有一定的治疗效果。结论:在本实验中对Movat染色的染液配置、染色时间、染色温度等进行细化和调整,为实验操作流程的规范化提供依据。在保证了实验统一性的基础上,可客观评估药物对动脉粥样硬化的治疗效果,同时因粥样斑块各结构成分颜色分明,利于镜下成像鉴别,可对各成分进行定量统计分析,为实验结果提供了可靠的实验数据。
ApoE knockout mice are one of the common models for the study of atherosclerosis. They can accelerate the formation of aortic atherosclerotic plaque under high fat diet. Atherosclerosis of the lesion components are smooth muscle cells, foam cells, cholesterol crystals, collagen fibers, elastic fibers and proteoglycans. Movat staining technology can visualize the pathological changes of these components on a slice. The standardized and detailed experimental procedures can ensure the consistency of the staining results and provide a clear and reliable experimental basis for the further study of atherosclerosis , So as to more objectively evaluate the experimental results. Methods: Seventy male Apo E mice were randomly divided into 8 groups: Apo E blank group, positive control group, treatment A high, medium and low dose groups, treatment B high, medium and low dose groups , And 6 C57 mouse control groups. After 26 weeks of high fat feeding, blood was taken and sacrificed. Take the heart, paraffin embedded sections after positioning. Batch dyeing, dyeing process to explore and refine the dye configuration, dyeing temperature, dyeing time and other conditions, standardize the experimental operation. Into the film after the histopathological observation, and collagen fibers, elastic fibers, proteoglycans, foam cells and other qualitative and quantitative analysis. Results: A refined and standardized Movat dyeing procedure was established through experiments. Light microscopic observation of the staining results showed that each structural component was clearly colored: collagen fibers and elastic fibers were yellow and black, proteoglycan was green, foam cells were lavender, myocardial smooth muscle was red. Atherosclerotic plaque lesions at a clear structure, the pathological manifestations of vascular smooth muscle proliferation, collagen fibers, smooth muscle cells and proteoglycans to form a fibrous cap, the fibrous cap formed under the foam cells and extends downward, a large number of lipid deposition, elastic fibers fracture. Compared with the blank group and the positive control group, the therapeutic drugs A and B both have a certain therapeutic effect on the severity of the disease and the progression of the disease. Conclusion: In this experiment, the configuration, dyeing time and dyeing temperature of Movat dyeing were refined and adjusted, which provided the basis for the standardization of experimental operation procedure. On the basis of ensuring the unification of experiments, the therapeutic effect of drugs on atherosclerosis can be objectively evaluated. At the same time, the structural components of atherosclerotic plaque are distinguished by the distinct colors, which are good for the identification of microscopic imaging. Quantitative and statistical analysis of each component can be carried out, The experimental results provide reliable experimental data.