Al-Obaidi Mazen M Jamil, Bahadoran Azadeh, Har Lee Sau, Mui Wang Seok, Rajarajeswaran Jayakumar, Zandi Keivan, Manikam Rishya, Sekaran Shamala Devi
Department of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
Institute of Medical Molecular Biotechnology, Faculty of Medicine, University Technology Mara, Selangor, Malaysia.
Virus Res. 2017 Apr 2;233:17-28. doi: 10.1016/j.virusres.2017.02.012. Epub 2017 Mar 6.
Japanese encephalitis (JE) is a neurotropic flavivirus that causes inflammation in central nervous system (CNS), neuronal death and also compromises the structural and functional integrity of the blood-brain barrier (BBB). The aim of this study was to evaluate the BBB disruption and apoptotic process in Japanese encephalitis virus (JEV)-infected transfected human brain microvascular endothelial cells (THBMECs). THBMECs were overlaid by JEV with different MOIs (0.5, 1.0, 5.0 and 10.0) and monitored by electrical cell-substrate impedance sensing (ECIS) in a real-time manner in order to observe the barrier function of THBMECs. Additionally, the level of 43 apoptotic proteins was quantified in the virally infected cells with different MOIs at 24h post infection. Infection of THBMEC with JEV induced an acute reduction in transendothelial electrical resistance (TEER) after viral infection. Also, significant up-regulation of Bax, BID, Fas and Fasl and down-regulation of IGFBP-2, BID, p27 and p53 were observed in JEV infected THBMECs with 0.5 and 10 MOIs compared to uninfected cells. Hence, the permeability of THBMECs is compromised during the JEV infection. In addition high viral load of the virus has the potential to subvert the host cell apoptosis to optimize the course of viral infection through deactivation of pro-apoptotic proteins.
日本脑炎(JE)是一种嗜神经性黄病毒,可引起中枢神经系统(CNS)炎症、神经元死亡,并损害血脑屏障(BBB)的结构和功能完整性。本研究的目的是评估日本脑炎病毒(JEV)感染的转染人脑血管内皮细胞(THBMECs)中的血脑屏障破坏和凋亡过程。用不同感染复数(MOI,0.5、1.0、5.0和10.0)的JEV感染THBMECs,并通过细胞-基质电阻抗传感(ECIS)进行实时监测,以观察THBMECs的屏障功能。此外,在感染后24小时对不同MOI的病毒感染细胞中的43种凋亡蛋白水平进行定量。JEV感染THBMECs后,病毒感染后跨内皮电阻(TEER)急剧下降。此外,与未感染细胞相比,在MOI为0.5和10的JEV感染的THBMECs中观察到Bax、BID、Fas和Fasl显著上调,IGFBP-2、BID、p27和p53下调。因此,在JEV感染期间,THBMECs的通透性受到损害。此外,高病毒载量有可能通过使促凋亡蛋白失活来颠覆宿主细胞凋亡,从而优化病毒感染过程。