基于组织病理学的大鼠脑干原发性功能损伤程度研究  

Histopathology-based primary functional impairment of brainstem in rats

作　　者：傅倩颖 石菁菁 王国祥 朱林娜 王讯 杨袖菊 刘盛雄 Fu Qian-ying;Shi Jing-jing;Wang Guo-xiang;Zhu lin-na;Wang Xun;Yang Xiu-ju;Liu Sheng-xiong(Department of Biomedical Engineering,College of Pharmacy and Bioengineering,Chongqing University of Technology,Chongqing 400054,China;Key Laboratory for Biorheological Science and Technology of Ministry of Education,State and Local Joint Engineering Laboratory for Vascular Implants,Bioengineering College of Chongqing University,Chongqing 400030,China)

机构地区：[1]重庆理工大学药学与生物工程学院,重庆400054 [2]重庆大学生物工程学院,重庆400030

出　　处：《四川生理科学杂志》2023年第6期941-947,共7页Sichuan Journal of Physiological Sciences

基　　金：国家重点研发项目(2016YFC0800702-3);重庆理工大学研究生教育高质量发展行动计划资助成果(gzlcx20222091)。

摘　　要：目的:利用自制改进的Marmarou模型建立不同损伤高度的弥漫性轴索损伤(Diffuse axonal injury,DAI)大鼠模型,探讨通过苏木精-伊红(Hematoxylin eosin staining,HE)染色和β-淀粉样前体蛋白(β-Amyloid precursor protein,β-APP)免疫组化互相验证结合的方式对大鼠脑干原发性功能损伤程度进行定量研究。方法:健康成年雄性(Sprague-dawley,SD)大鼠24只,随机分为0 m正常组、1.25 m致伤组和2.25 m致伤组,采用改进的经典Marmarou模型制作大鼠脑干部位的DAI模型,正常组不进行打击。在致伤后进行过量麻醉取材制作HE染色切片和β-APP免疫组化染色切片,其中对HE染色切片计算脑干感兴趣区域内出血占比、脑水肿个数和锥体束出血占比;对β-APP免疫组化切片测定脑干感兴趣区域内的阳性细胞面积和累计光密度值(Integral optical density,IOD)。将不同损伤程度脑组织切片的HE染色分析与β-APP免疫组化染色结合做相关性分析。结果:不同致伤高度组的HE染色切片在光镜下可见神经束疏散、紊乱,血管淤滞,出现不同程度的散点状或长条状出血,细胞核显示不清,突触结构模糊不清等经典DAI病理特征;β-APP免疫组化出现轻度轴索损伤变化。测定脑干感兴趣区域内β-APP的阳性细胞面积和IOD值与HE染色的出血面积占比和锥体束出血面积占比呈正相关。结论:采用HE染色计算的大鼠脑干位置出血面积占比和锥体束区域出血面积占比和β-APP免疫组化计算的阳性细胞面积和IOD值可以定量区分大鼠脑干损伤程度,为判断大鼠脑干原发性功能损伤提供新角度,有助于为交通事故和法医检测等脑损伤机制的研究提供实验参考。Objective:The self-made and improved Marmarou model was used to establish a rat model of diffuse axonal injury(DAI)with different injury heights,and the degree of primary functional impairment in rats through hematoxylin-eosin staining(HE)andβ-Amyloid Precursor Protein(β-APP)immunohistochemistry staining were used to quantitatively study the degree of primary functional impairment in rats.Methods:There were 24 healthy adult male(Sprague-Dawley,SD)rats,randomly divided into 0m control group,1.25m experimental group and 2.25m experimental group,and the improved classic Marmarou model was used to make DAI model of rat brainstem,and the control group did not strike.HE stained sections andβ-APP immunohistochemical staining sections were prepared by excessive anesthesia after injury,and the proportion of hemorrhage,cerebral edema and pyramidal tract hemorrhage in the brainstem region of interest was calculated for HE-stained sections.Measure positive cell area and integral optical density(IOD)value within the brainstem region of interest onβ-APP immunohistochemical sections.HE staining analysis of brain tissue sections with different degrees of injury was combined withβ-APP immunohistochemical staining for correlation analysis.Results:HE stained sections of different injury height groups showed nerve bundle evacuation,disorder,vascular stasis,scattered or long hemorrhage of different degrees,unclear nuclei display,and ambiguous synaptic structure under light microscopy.β-APP immunohistochemistry showed mild axonal injury changes.The positive cell area and IOD value ofβ-APP in the brainstem region of interest were positively correlated with the proportion of bleeding area and pyramidal tract bleeding area in HE staining.Conclusion:The proportion of bleeding area at the brainstem position and the proportion of bleeding area in the pyramidal tract region calculated by HE staining,and the positive cell area and IOD value calculated byβ-APP immunohistochemistry can quantitatively distinguish the degree of brainstem

关 键 词：组织病理学 大鼠脑干 原发性损伤 功能损伤程度 

分 类 号：R33[医药卫生—人体生理学] Q4[医药卫生—基础医学]