Yamamoto Masayoshi, Umebashi Katsuyuki, Tokito Akinori, Imamura Junichi, Jougasaki Michihisa
Institute for Clinical Research, National Hospital Organization Kagoshima Medical Center, Kagoshima, Japan; and.
Neurohumoral Biology, Cooperative Department of Innovative Medicine, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, Japan.
Am J Physiol Regul Integr Comp Physiol. 2017 Sep 1;313(3):R272-R279. doi: 10.1152/ajpregu.00435.2016. Epub 2017 Jun 21.
Although interleukin-33 (IL-33), a member of the IL-1 cytokine family, plays proinflammatory roles in immune cells as an "alarmin," little is known regarding the biological actions of IL-33 on vascular endothelial cells. To investigate the effects of IL-33 on vascular endothelial cells, we first screened the IL-33-regulated proteins in human umbilical vein endothelial cells (HUVECs) using a dot blot array and observed that IL-33 markedly increased growth-regulated oncogene-α (GRO-α), a chemokine that is also known as chemokine (C-X-C motif) ligand 1 (CXCL1). Real-time reverse transcription PCR and ELISA demonstrated that IL-33 induced GRO-α expression and secretion in HUVECs in a dose- and a time-dependent manner. Western immunoblot assay revealed that IL-33 activated the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) and c-Jun NHterminal kinase (JNK). In addition, translocation of nuclear factor-κB (NF-κB) p65 to the nucleus of HUVECs was observed by IL-33 stimulation. Furthermore, treatment with pharmacological inhibitors against ERK1/2 (PD98059), JNK (SP600125), or NF-κB (BAY11-7085) significantly suppressed IL-33-induced GRO-α gene expression and secretion from HUVECs. Moreover, immunohistochemical staining demonstrated that IL-33 and GRO-α coexpressed in the endothelium of human carotid atherosclerotic plaque. Taken together, the present study indicates that IL-33 localized in the human atherosclerotic plaque increases GRO-α mRNA expression and protein secretion via activation of ERK1/2, JNK, and NF-κB in HUVECs, suggesting that IL-33 plays an important role in the pathophysiology and development of atherosclerosis.
白细胞介素-33（IL-33）是白细胞介素-1细胞因子家族的成员，作为一种“警报素”在免疫细胞中发挥促炎作用，但关于IL-33对血管内皮细胞的生物学作用知之甚少。为了研究IL-33对血管内皮细胞的影响，我们首先使用斑点印迹阵列筛选了人脐静脉内皮细胞（HUVECs）中IL-33调节的蛋白质，并观察到IL-33显著增加了生长调节致癌基因-α（GRO-α），一种也被称为趋化因子（C-X-C基序）配体1（CXCL1）的趋化因子。实时逆转录PCR和ELISA表明，IL-33以剂量和时间依赖性方式诱导HUVECs中GRO-α的表达和分泌。蛋白质免疫印迹分析显示，IL-33激活了细胞外信号调节激酶1/2（ERK1/2）和c-Jun氨基末端激酶（JNK）的磷酸化。此外，通过IL-33刺激观察到核因子-κB（NF-κB）p65向HUVECs细胞核的转位。此外，用针对ERK1/2（PD98059）、JNK（SP600125）或NF-κB（BAY11-7085）的药理抑制剂处理显著抑制了IL-33诱导的HUVECs中GRO-α基因的表达和分泌。此外，免疫组织化学染色表明，IL-33和GRO-α在人颈动脉粥样硬化斑块的内皮中共表达。综上所述，本研究表明，定位于人动脉粥样硬化斑块中的IL-33通过激活HUVECs中的ERK1/2、JNK和NF-κB增加GRO-α mRNA表达和蛋白质分泌，提示IL-33在动脉粥样硬化的病理生理学和发展中起重要作用。
