改良的动脉内皮细胞铺片技术-医学形态学与医学机能学的实验教学融合设计

A modified technique for arterial endothelium preparation–an integrated approach in teaching of Medical Morphology and Medical Functional Experiment

目的改进动脉内皮细胞铺片技术,探索并创建一种将医学形态学和医学机能学实验相结合的实验教学内容。方法健康雄性SD大鼠20只分别给予高脂饮食和正常饮食,8周后检测各组体重、血脂、胆固醇水平和肝功相关指标的变化;分离大鼠主动脉,简化玻片包被、组织脱水及烤片等操作流程,制作动脉内皮细胞铺片,HE染色或联合免疫组织化学技术观察实验结果。上述实验流程由学生在指导下分4次课完成。结果与正常饮食组大鼠相比,高脂组大鼠总胆固醇(TC)、甘油三酯(TG)和低密度脂蛋白胆固醇(LDL-C)明显升高,高密度脂蛋白胆固醇(HDL-C)明显降低,血清谷草转氨酶(AST)、谷丙转氨酶(ALT)和碱性磷酸酶(ALP)水平明显增高;改进动脉内皮细胞铺片技术,学生能在预定学时内观察到高脂组大鼠内皮细胞数量增多,排列紊乱,形态不规则,胞核大小和染色深浅不一,内皮素(ET-1)阳性细胞率显著降低。结论高脂饲养的大鼠血脂指标紊乱,伴随不同程度的肝功能损坏和动脉内皮细胞损伤。改良后的动脉内皮铺片技术,将机能学、形态学变化与疾病的发生、发展和转归有机地融为一体,有助于建立学生的医学整体观和临床思维。

Objective To improve the technique of artery endothelium preparation,explore and establish an integrated teaching content of Medical Morphology and Medical Functional Experiment.Methods 20 healthy male SD rats were divided into control group(fed a normal diet)and hyperlipidic group(fed a hyperlipidic diet).Body weight,blood lipids and liver functions of both groups were measured after 8 weeks.Rat aortae were isolated;arterial endothelial cells were prepared by simplified processes of coating,dehydration and slides drying;and then results were observed by HE and/or immunohistochemical staining.The above experimental process was completed by students in 4 lessons under the guidance.Results Rats in hyperlipidic group showed increased serum TC,TG,LDL-C,AST,ALT,ALP and decreased HDL-C concentrations compared with the control group,while the body weight between the control group and hyperlipidic group showed no significant difference.Modified process of artery endothelium preparation technique was convenient for the students to observe the morphological injury of the arterial endothelium of hyperlipidic group.Within scheduled hours,students observed that the number of endothelial cells in the hyperlipidic group rats increased,the arrangement was disordered,the morphology was irregular,the nuclei size and staining depth were different,and the rate of endothelin(ET-1)-positive cells was significantly reduced.Conclusions Hyperlipidic rats showed dyslipidemia,liver damage and artery endothelium injury.Functional and morphological changes were integrated with the occurrence and development of disease by modified artery endothelium preparation technique,which was helpful for medical students to establish the holistic view of medicine and clinical thinking ability.