摘要
目的观察创伤性脑损伤(TBI)大鼠血脑屏障(BBB)中紧密连接蛋白闭合蛋白(Occludin)、闭锁小带蛋白1(ZO-1)的表达及变化趋势,探讨大麻二酚(CBD)对BBB的干预作用。方法采用改良Feeney自由落体法制备大鼠TBI模型,将大鼠随机分为3组:假手术组(Sham组)、模型组(TBI+vehicle组)和CBD干预组(TBI+CBD组),每组24只;每个组又分为损伤后8 h、1、2、3、5和7 d共6个时间点,通过免疫组化和免疫荧光染色,Western blot检测与BBB通透性密切相关的Occludin和ZO-1在不同时间点的表达情况;通过荧光素钠实验检测BBB通透性。结果免疫组化实验结果表明,与假手术组相比,TBI后随着时间推移Occludin和ZO-1蛋白阳性表达减少(P<0.05),2 d达到最低;CBD干预1 d后Occludin和ZO-1蛋白表达水平均上调(P<0.05);免疫荧光染色实验与Western blot结果趋势近似,与假手术组相比,TBI后Occludin和ZO-1荧光表达强度及蛋白表达量减少(P<0.05),CBD干预2 d后Occludin和ZO-1表达水平上调(P<0.05);荧光素钠实验结果表明,TBI后脑组织BBB完整性遭到破坏,通透性升高(P<0.01),CBD干预后BBB通透性下降(P<0.05)。结论TBI后紧密连接蛋白Occludin和ZO-1表达下降,BBB通透性升高,CBD干预可逆转TBI对BBB的破坏。
Objective To observe the expression and trends of tight junction proteins Occludin and zonula occluden-1(ZO-1)in blood-brain barrier(BBB)of rats with traumatic brain injury(TBI),and to explore the intervention effect of cannabidiol(CBD)on the BBB.Methods The TBI model of rat was prepared by modified“Feeney free fall method”and randomly divided into three groups:the sham-operated group(Sham group),the model group(TBI+vehicle group)and the CBD intervention group(TBI+CBD group),with 24 rats in each group.Each group was subdivided into six time points:8 h,1,2,3,5 and 7 d after injury.The expression of Occludin and ZO-1,which are closely related to the permeability of BBB,was detected by immunohistochemistry,immunofluorescence staining and Western blot at different points.The permeability of BBB was detected by sodium fluorescein assay.Results The results of immunohistochemistry showed that compared with Sham group,the positive expression of Occludin and ZO-1 decreased with time after brain trauma(P<0.05),and both reached the lowest level at 2 d.The expression levels of Occludin and ZO-1 were up-regulated after 1 d of CBD intervention(P<0.05).Immunofluorescence staining showed a similar trend to Western blot results,with Occludin and ZO-1 fluorescence expression intensity and protein expression reduced after TBI compared with Sham group(P<0.05).And the expression levels of Occludin and ZO-1 were up-regulated after 2 d of CBD intervention(P<0.05).The results of fluorescein sodium experiment showed that the BBB integrity of brain tissue was destroyed after TBI,and the permeability increased after TBI(P<0.01).The permeability of BBB decreased after CBD intervention(P<0.05).Conclusion The expression of tight junction proteins Occludin and ZO-1 decreases after TBI,and the permeability of BBB is disrupted,and CBD intervention reverses the disruption of the BBB by TBI.
作者
李佳丽
曹艳
凌腾晗
尹爱平
李恒希
李经辉
张瑞林
吴海鹰
李坪
Li Jiali;Cao Yan;Ling Tenghan;Yin Aiping;Li Hengxi;Li Jinghui;Zhang Ruilin;Wu Haiying;Li Ping(Dept of Human Anatomy and Histology&Embryology,Kunming Medical University,Kunming 650500;Dept of Neurosurgery,First Affiliated Hospital of Kunming Medical University,Kunming 650032;School of Forensic Medicine,Kunming Medical University,Kunming 650500;Dept of Emergency,First Affiliated Hospital of Kunming Medical University,Kunming 650032)
出处
《安徽医科大学学报》
CAS
北大核心
2024年第3期478-483,共6页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金(编号:82060241、82260387)
云南省科技厅计划项目(编号:202101AY070001-002)
昆明医科大学研究生创新基金(编号:2023S133)
