Nishihara Hideaki, Shimizu Fumitaka, Sano Yasuteru, Takeshita Yukio, Maeda Toshihiko, Abe Masaaki, Koga Michiaki, Kanda Takashi
Department of Neurology and Clinical Neuroscience, Yamaguchi University Graduate School of Medicine, Ube, Japan.
PLoS One. 2015 Mar 16;10(3):e0121488. doi: 10.1371/journal.pone.0121488. eCollection 2015.
Effect of fingolimod in multiple sclerosis (MS) is thought to involve the prevention of lymphocyte egress from lymphoid tissues, thereby reducing autoaggressive lymphocyte infiltration into the central nervous system across blood-brain barrier (BBB). However, brain microvascular endothelial cells (BMECs) represent a possible additional target for fingolimod in MS patients by directly repairing the function of BBB, as S1P receptors are also expressed by BMECs. In this study, we evaluated the effects of fingolimod on BMECs and clarified whether fingolimod-phosphate restores the BBB function after exposure to MS sera.
Changes in tight junction proteins, adhesion molecules and transendothelial electrical resistance (TEER) in BMECs were evaluated following incubation in conditioned medium with or without fingolimod/fingolimod-phosphate. In addition, the effects of sera derived from MS patients, including those in the relapse phase of relapse-remitting (RR) MS, stable phase of RRMS and secondary progressive MS (SPMS), on the function of BBB in the presence of fingolimod-phosphate were assessed.
Incubation with fingolimod-phosphate increased the claudin-5 protein levels and TEER values in BMECs, although it did not change the amount of occludin, ICAM-1 or MelCAM proteins. Pretreatment with fingolimod-phosphate restored the changes in the claudin-5 and VCAM-1 protein/mRNA levels and TEER values in BMECs after exposure to MS sera.
Pretreatment with fingolimod-phosphate prevents BBB disruption caused by both RRMS and SPMS sera via the upregulation of claudin-5 and downregulation of VCAM-1 in BMECs, suggesting that fingolimod-phosphate is capable of directly modifying the BBB. BMECs represent a possible therapeutic target for fingolimod in MS patients.
芬戈莫德在多发性硬化症(MS)中的作用被认为涉及防止淋巴细胞从淋巴组织中逸出,从而减少自身攻击性淋巴细胞通过血脑屏障(BBB)浸润到中枢神经系统。然而,脑微血管内皮细胞(BMECs)可能是芬戈莫德在MS患者中的另一个潜在靶点,因为BMECs也表达S1P受体,芬戈莫德可通过直接修复BBB功能发挥作用。在本研究中,我们评估了芬戈莫德对BMECs的影响,并阐明了磷酸芬戈莫德在暴露于MS血清后是否能恢复BBB功能。
在含有或不含有芬戈莫德/磷酸芬戈莫德的条件培养基中孵育后,评估BMECs中紧密连接蛋白、黏附分子和跨内皮电阻(TEER)的变化。此外,还评估了来自MS患者的血清,包括复发缓解型(RR)MS复发期、RRMS稳定期和继发进展型MS(SPMS)患者的血清,在磷酸芬戈莫德存在下对BBB功能的影响。
用磷酸芬戈莫德孵育可增加BMECs中claudin-5蛋白水平和TEER值,尽管它没有改变occludin、ICAM-1或MelCAM蛋白的含量。磷酸芬戈莫德预处理可恢复BMECs在暴露于MS血清后claudin-5和VCAM-1蛋白/ mRNA水平以及TEER值的变化。
磷酸芬戈莫德预处理可通过上调BMECs中claudin-5和下调VCAM-1来防止RRMS和SPMS血清引起的BBB破坏,这表明磷酸芬戈莫德能够直接调节BBB。BMECs可能是芬戈莫德在MS患者中的一个治疗靶点。