Department of Biomedical Science, Jungwon University, 85 Munmu-ro, Goesan-eup, Goesan-gun, Chungbuk 28024, Korea.
Division of Cardiology, Samsung Medical Center, Sungkyunkwan University School of Medicine 50 Irwon dong, Gangnam-gu, Seoul 06351, Korea.
Int J Mol Sci. 2021 Mar 17;22(6):3053. doi: 10.3390/ijms22063053.
Coxsackievirus and adenovirus receptor (CAR) is present in epithelial and vascular endothelial cell junctions. We have previously shown a hemorrhagic phenotype in germ-line CAR knock-out mouse embryos; we have also found that CAR interacts with ZO-1 and β-catenin. However, the role of CAR in vascular endothelial junction permeability has not been proven. To understand the roles of CAR in the vascular endothelial junctions, we generated endothelium-specific CAR knockout (CAR-eKO) mice. In the absence of CAR, the endothelial cell layer showed an increase in transmembrane electrical resistance (TER, Ω) and coxsackievirus permeability. Evans blue dye and 70 kDa dextran-FITC were delivered by tail vein injection. We observed increased vascular permeability in the hearts of adult CAR-eKO mice compare with wild-type (WT) mice. There was a marked increase in monocyte and macrophage penetration into the peritoneal cavity caused by thioglycolate-induced peritonitis. We found that CAR ablation in endothelial cells was not significantly increased coxsackievirus B3 (CVB3) induced myocarditis in murine model. However, tissue virus titers were significantly higher in CAR-eKO mice compared with WT. Moreover, CVB3 was detected in the brain of CAR-eKO mice. Endothelial CAR deletion affects the expression of major endothelial junction proteins, such as cadherin and platelet endothelial cell adhesion molecule-1 (PECAM-1) in the cultured endothelial cells as well as liver vessel. We suggest that CAR expression is required for normal vascular permeability and endothelial tight junction homeostasis. Furthermore, CVB3 organ penetration and myocarditis severities were dependent on the endothelial CAR level.
柯萨奇病毒和腺病毒受体 (CAR) 存在于上皮细胞和血管内皮细胞连接处。我们之前曾在 CAR 基因敲除小鼠胚胎中观察到出血表型;我们还发现 CAR 与 ZO-1 和 β-连环蛋白相互作用。然而,CAR 在血管内皮连接通透性中的作用尚未得到证实。为了了解 CAR 在血管内皮连接中的作用,我们生成了内皮细胞特异性 CAR 敲除 (CAR-eKO) 小鼠。在没有 CAR 的情况下,内皮细胞层的跨膜电阻 (TER,Ω) 和柯萨奇病毒通透性增加。通过尾静脉注射 Evans 蓝染料和 70 kDa 葡聚糖-FITC。与野生型 (WT) 小鼠相比,我们观察到成年 CAR-eKO 小鼠心脏的血管通透性增加。在由巯基醋酸盐诱导的腹膜炎引起的单核细胞和巨噬细胞渗透到腹腔中,观察到明显增加。我们发现内皮细胞中 CAR 的缺失并没有显著增加柯萨奇病毒 B3 (CVB3) 在小鼠模型中诱导的心肌炎。然而,与 WT 相比,CAR-eKO 小鼠的组织病毒滴度明显更高。此外,在 CAR-eKO 小鼠的大脑中检测到 CVB3。内皮细胞 CAR 缺失会影响培养的内皮细胞以及肝血管中主要内皮连接蛋白的表达,如钙粘蛋白和血小板内皮细胞黏附分子-1 (PECAM-1)。我们认为 CAR 表达对于正常的血管通透性和内皮紧密连接稳态是必需的。此外,CVB3 器官穿透和心肌炎严重程度取决于内皮细胞 CAR 水平。