HMGB1 contributes to the irradiation-induced endothelial barrier injury through receptor for advanced glycation endproducts (RAGE)
HMGB1 contributes to the irradiation-induced endothelial barrier injury through receptor for advanced glycation endproducts (RAGE)
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HMGB1 通过晚期糖基化终末产物受体 (RAGE) 促进辐射诱导的内皮屏障损伤
DOI：
10.1002/jcp.26341
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发表时间：
2018
影响因子：
5.6
通讯作者：
Yu Yangsheng
中科院分区：
文献类型：
--
作者：
Zhou Haihong;Jin Congli;Cui Lili;Xing Huaijie;Liu Jun;Liao Wang;Liao Haojie;Yu Yangsheng
本研究旨在探讨高迁移率族蛋白-1(HMGB1)和晚期糖基化终产物受体(RAGE)是否参与辐射致内皮细胞屏障损伤及其机制。用bEnd.3细胞建立内皮屏障损伤模型。用荧光素钠(Na-F)渗透试验检测内皮细胞屏障的通透性，并通过不同的照射剂量(5、10、15、20 Gy5、10、15、20 Gy4个剂量)确定可诱导通透性转变的照射剂量。用四甲基偶氮唑盐比色法检测不同浓度HMGB1、甘草酸(GA，HMGB1的特异性抑制剂)和FPS-ZM1(RAGE V结构域介导的配体结合的血脑屏障阻断剂)对细胞存活率的影响。采用定量逆转录聚合酶链式反应、免疫印迹和免疫荧光法检测HMGB1、RAGE、MAPK信号通路相关分子、MMP2、MMP9、ZO 1、CLADIN5在不同处理组中的表达。细胞接种于Transwell板后4~7d内具有稳定的内皮屏障功能。内皮屏障通透性在至少10 Gy射线照射下发生改变。辐射和HMGB1单独处理均能提高透性。照射后HMGB1和RAGE表达增加，MAPK信号通路激活。同时，MMP2和MMP9表达增强，紧密连接蛋白ZO 1和CLADIN5表达降低。辐射可通过促进HMGB1和RAGE的表达激活MAPK信号通路，进而导致内皮屏障损伤，改变其通透性。
This study aimed to investigate whetherHMGB1(high mobility group box‐1 protein) and receptor for advanced glycation end products (RAGE) were involved in the irradiation‐induced endothelial barrier damage and their mechanism. We constructed the damage model of endothelium barrier model with bEnd.3 cells. The permeability of endothelial barrier was detected by sodium fluorescein (Na‐F) permeation test, and the irradiation dose which could induce permeability transition was determined by being exposed to different irradiation doses (5, 10, 15, 20 Gy). MTT assay was applied to detect cell viability under different concentrations ofHMGB1, glycyrrhizic acid (GA, a specific inhibitor ofHMGB1), and FPS‐ZM1 (a blood‐brain‐barrier permeant blocker of RAGE V domain‐mediated ligand binding). The expression ofHMGB1, RAGE, and related molecules involved in MAPK signaling pathway, MMP‐2, MMP‐9, ZO‐1, and claudin 5 of differently treated groups were measured by qRT‐PCR, western blot, and immunofluorescence. Cells possessed stable endothelial barrier function on 4–7 days after seeded on transwell plates. The permeability of endothelial barrier would change under at least 10 Gy radiation. Both radiation andHMGB1treatment alone could improve the permeability. After irradiation, the expressions ofHMGB1and RAGE increased and MAPK signal pathway was activated. Meanwhile, MMP‐2 and MMP‐9 were overexpressed, while the expression of tight junction proteins ZO‐1 and claudin 5 was decreased. Radiation could activate MAPK signaling pathway through promoting the expression ofHMGB1and RAGE, which further led to endothelial barrier injury and changed its permeability.