摘要
目的通过颈内动脉单侧、单次注射月桂酸钠,建立脑小血管病(cerebral small vessel disease,CSVD)大鼠模型,并通过检测行为学、血清相关指标、脑梗死体积、脑微血管密度、血流动力学、脑组织病理学及血脑屏障相关指标,评估模型的有效性。方法将SPF级雄性SD大鼠随机分为对照组(Control组)和模型组(Model组),每组6只。模型组经颈内动脉,单次注射100μL的月桂酸钠(2 g/L),对照组行相同手术,注射等体积的生理盐水。利用Longa评分及姿势反射实验进行大鼠神经行为学评估;ELISA检测大鼠血清同型半胱氨酸(homocysteine,HCY)含量;核磁共振成像(magnetic resonance imaging,MRI)检测脑梗死体积、脑血管成像观察脑血管密度改变;超声检测颈动脉血管阻力指数(resistance index,RI)及血流灌注指数(perfusion index,PI);苏木素-伊红(HE)染色观察大脑组织病理变化;免疫组化(IHC)检测脑组织中脑微血管密度CD31及紧密连接蛋白(ZO-1、Occludin)的表达。结果与对照组相比,模型组Longa评分与姿势反射评分均显著升高(P<0.05),脑梗死体积显著增加(P<0.05),脑血管密度明显下降,颈动脉RI、PI值及血清中HCY的含量均显著升高(P<0.05);HE染色结果发现模型组的脑皮质区神经元核固缩,血管周围间隙变大;免疫组化结果显示脑皮质区CD31、ZO-1、Occludin的表达显著减少(P<0.05)。结论通过颈内动脉单侧、单次注射高浓度月桂酸钠可以快速、有效地建立CSVD模型;该方法建立的CSVD大鼠模型,存在神经行为学异常、脑梗死、脑供血不足、血管密度减少和血脑屏障破坏现象,可作为CSVD研究的有效动物模型。
Objective A rat model of cerebral small vessel disease(CSVD)was established by unilateral injection of a single dose of sodium laurate into the internal carotid artery.The effectiveness of the model was assessed by behavior scoring and analysis of serum-related indicators,cerebral infarction volume,cerebral microvascular density,hemodynamics,brain histopathology and the expression of blood-brain barrier(BBB)-related proteins.Methods SPF-grade male SD rats were divided randomly into a control group and a model group(n=6 per group).The model group received a single injection of 100μL of sodium laurate(2 g/L)via the internal carotid artery,while the control group underwent the same surgical procedure but received an equal volume of saline.Neurobehavioral assessments were conducted using the Longa score and postural reflex test.Serum homocysteine(HCY)levels were measured by enzyme-linked immunosorbent assay.Cerebral infarction volume was detected by magnetic resonance imaging and changes in cerebral vascular density were observed by cerebrovascular imaging.The resistance index(RI)and perfusion index(PI)were measured by ultrasonography.Histopathological changes in brain tissue were evaluated by hematoxylin and eosin(HE)staining.Expression of the cerebral microvascular marker CD31 and tight junction proteins ZO-1 and Occludin in brain cortex tissue were detected by immunohistochemical staining.Results The Longa score,postural reflex score(P<0.05),and cerebral infarction volume were significantly increased(P<0.05)while the cerebral vascular density was decreased in the model group compared with the control group.Serum HCY levels,carotid RI,and PI values were all significantly increased in the model group(P<0.05).HE staining revealed solidified neuronal nuclei and enlarged perivascular spaces in the brain cortex in the model group.Immunohistochemical staining revealed that CD31,ZO-1,and Occludin expression were significantly reduced in the brain cortex in the model group compared with the control group(P<0.05).Conclusions A rat model of CSVD can be established rapidly and effectively by a single unilateral injection of high-concentration sodium laurate via the internal carotid artery.This model is characterized by neurobehavioral abnormalities,cerebral infarction,insufficient cerebral blood supply,reduced vascular density,and disruption of the BBB,suggesting that it may serve as an effective rat model for the study of CSVD.
作者
陈炎森
林浩伟
张予菲
林雨星
曹长源
赖可欣
吴钰婷
蔡巧燕
张铃
CHEN Yansen;LIN Haowei;ZHANG Yufei;LIN Yuxing;CAO Changyuan;LAI Kexin;WU Yuting;CAI Qiaoyan;ZHANG Ling(College of Integrative Medicine/Academy of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;College of Life Sciences and Food Engineering,Shanxi Xueqian Normal University,Xi’an 710100,China;College of Traditional Chinese Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;Fujian Key Laboratory of Integrative Medicine on Geriatrics,Fuzhou 350122,China)
出处
《中国实验动物学报》
北大核心
2025年第6期779-789,共11页
Acta Laboratorium Animalis Scientia Sinica
基金
国家自然科学基金(82374282)
福建省自然科学基金(2022J01876,2022J01875)
国家级大学生创新创业训练计划项目(202310393006)
福建中医药大学青年科技创新培育计划项目(XQC2024004)。
