Icam-1介导广州管圆线虫感染小鼠血脑屏障破损的研究  

Icam-1 leads to blood-brain barrier disruption in mice caused by Angiostrongylus cantonensis infection

作　　者：覃秋华 韦航 吕志跃 QIN Qiuhua;WEI Hang;LU Zhiyue(Department of Parasitology,Zhongshan School of Medicine,Sun Yat-sun University,Guangzhou,Guangdong 510080,China)

机构地区：[1]中山大学中山医学院寄生虫教研室,广东广州510080

出　　处：《热带医学杂志》2023年第4期429-434,F0002,458,共8页Journal of Tropical Medicine

基　　金：国家自然科学基金(82072303);广东省科技计划项目(2019B030316025);广东省自然科学基金(2019A1515011541);国家重点研发计划项目(2021YFC23008001,2021YFC23008002);海南省重点研发计划项目(ZDYF2020120)。

摘　　要：目的探究广州管圆线虫感染对小鼠血脑屏障破损的分子机制。方法构建广州管圆线虫病Balb/c小鼠模型,通过HE染色检查小鼠脑部病理损伤情况,免疫荧光检测白蛋白(Alb)渗漏情况。分离脑组织,提取RNA和蛋白质,RT-qPCR检测小鼠血脑屏障破坏相关分子基因表达水平变化;Western blot检测小鼠细胞间黏附分子-1(Icam-1)蛋白水平变化。免疫荧光检测小鼠Icam-1表达情况,Icam-1与Alb共定位情况。构建广州管圆线虫感染Balb/c小鼠后抑制剂A205804治疗模型,通过伊文思蓝渗透性实验检测感染小鼠、正常小鼠和抑制剂治疗小鼠血脑屏障渗透性情况。HE染色检测A205804对感染小鼠脑部病理损伤情况。通过免疫荧光检测A205804对感染小鼠Icam-1表达的抑制情况以及白蛋白渗漏情况。结果随着感染时间增加,脑组织开始出现明显的脑膜增厚、炎性细胞增多。免疫荧光结果显示,在感染后第21天看到Alb渗漏到血管外部。RT-qPCR显示,与未感染小鼠相比,感染小鼠Icam-1表达显著升高,差异有统计学意义(P<0.05)。经WB结果验证,与未感染小鼠相比,感染小鼠Icam-1含量升高,差异有统计学意义(P<0.05)。免疫荧光染色显示,小鼠在感染第21天Icam-1在脉络从与脑膜处表达显著增加,同时Alb和Icam-1均在脑膜处共表达。给予A205804后,伊文思蓝在脑组织中的渗漏量明显减少。HE染色显示脑膜处的炎性细胞数量减少。免疫荧光检测显示小鼠脑组织中Icam-1和白蛋白的表达减少。结论广州管圆线虫感染可诱导Balb/c小鼠通过上调Icam-1导致血脑屏障破坏,而Icam-1抑制剂A205804可以降低血脑屏障破坏程度。Objective To study the mechanism of destruction of blood-brain barrier(BBB)in mice infected with Angiostrongylus cantonensis.Methods Balb/c mouse model of A.cantonensis infection was established.HE staining was used to examine the pathological damage of the brain,and immunofluorescence was used to detect the leakage of albumin(Alb).Brain tissue was isolated,RNA and protein were extracted,and RT-qPCR was used to detect the changes in the expression levels of genes related to blood-brain barrier destruction in mice.The expression of Icam-1 protein was detected by Western blot.Immunofluorescence was used to detect the expression of intercellular cell adhesion molecule-1(Icam-1)and the co-localization of Icam-1 and Alb.A Balb/c mouse model of A.cantonensis infection was constructed and treated with inhibitor A205804.The permeability of BBB in the infected mice,normal mice and inhibitor treated mice was detected by Evans blue permeability test.HE staining was used to detect the pathological changes in the brain of infected mice treated with A205804.Immunofluorescence was used to detect the inhibitory effect of A205804 on Icam-1 expression and albumin leakage in the infected mice.Results With the increase of infection time,the brain tissue began to appear obvious meningeal thickening and inflammatory cells increased.The results of immunofluorescence showed that Alb leaked to the outside of the blood vessels on the 21 d infection.RT-qPCR showed that Icam-1 expression was significantly increased in the infected mice,the difference was statistically significant(P0.05).As verified by WB results,compared with uninfected mice,the Icam-1 expression of infected mice was increased,the difference was statistically significant(P0.05).Immunofluorescence staining showed that the expression of Icam-1 in the choroid plexus and meningeal was significantly increased on the 21 d of infection,and Alb and Icam-1 were co-expressed in the meningeal.After administration of A205804,the amount of Evans blue leakage in brain tissue was signific

关 键 词：广州管圆线虫 血脑屏障 ICAM-1 A205804 

分 类 号：R383.19[医药卫生—医学寄生虫学]