Bone Morphogenetic Protein-2 Induces Non-Canonical Inflammatory and Oxidative Pathways in Human Retinal Endothelial Cells.
Bone Morphogenetic Protein-2 Induces Non-Canonical Inflammatory and Oxidative Pathways in Human Retinal Endothelial Cells.
复制标题
DOI：
10.3389/fimmu.2020.568795
复制
发表时间：
2020
影响因子：
7.3
通讯作者：
Ibrahim AS
中科院分区：
文献类型：
--
作者：
Al-Shabrawey M;Hussein K;Wang F;Wan M;Elmasry K;Elsherbiny N;Saleh H;Yu PB;Tawfik A;Ibrahim AS
糖尿病视网膜病变（DR）的发病机制尚不完全清楚。我们之前证明了实验性糖尿病患者和糖尿病患者视网膜中视网膜骨形态发生蛋白-2 （BMP2）的上调。本研究旨在探讨非典型炎症途径在bmp2诱导的视网膜内皮细胞（REC）屏障功能障碍中的作用。为此，我们使用RT-PCR和western blotting技术评估了BMP2信号组分（BMP2、BMP4、BMP受体）、VEGF、磷酸化p38 MAPK和NFκB以及氧化应激标志物在培养的人视网膜内皮细胞（HRECs）中受到BMP2 （50ng/ml）长达24小时的影响。此外，我们还检测了高糖（HG、30mM d -葡萄糖）对HRECs中BMP2及其下游基因表达的影响。H2-DCF是一种荧光染料，测量细胞活性氧（ROS）水平，用于测量BMP2的促氧化作用。此外，我们通过使用电-底阻抗传感（ECIS）测量跨内皮细胞电阻（TER）来评估抑制p38和VEGF信号对bmp2诱导的HRECs屏障功能障碍的影响。我们还测试了HG在存在或不存在BMP2信号抑制剂的情况下对HRECs屏障完整性的影响。我们的数据显示，BMP2和高糖上调BMP信号通路中的BMP组分（SMAD效应物、BMP受体和TGFβ配体本身），诱导p38 MAPK和NFκB的磷酸化，并导致NFκB的核易位。抑制p38或NFκB可减弱bmp2诱导的HRECs中VEGF表达和屏障功能障碍。此外，抑制VEGFR2可减轻bmp2诱导的屏障功能障碍。此外，BMP2诱导ROS的产生和内皮型一氧化氮合酶（eNOS）在HRECs中的表达和活性。最后，HG上调HRECs中的BMP2及其下游基因（SMAD、BMP4、ALKs和TGF-β）， BMP2抑制剂可减轻HG诱导的HRECs屏障功能障碍。我们的研究结果表明，除了常规的SMAD信号外，BMP2还通过激活p38/NFκB通路诱导HRECs中的非典型炎症通路，导致VEGF上调和HRECs的破坏。抑制BMP2信号是一种潜在的治疗干预措施，可以保护内皮细胞屏障功能。
The mechanisms of diabetic retinopathy (DR), are not yet fully understood. We previously demonstrated an upregulation of retinal bone morphogenetic protein-2 (BMP2) in experimental diabetes and in retinas of diabetic human subjects. The purpose of current study was to investigate the role of non-canonical inflammatory pathway in BMP2-induced retinal endothelial cell (REC) barrier dysfunction. For this purpose, we used RT-PCR and western blotting to evaluate the levels of BMP2 signaling components (BMP2, BMP4, BMP receptors), VEGF, phosphorylated p38 MAPK and NFκB, and oxidative stress markers in cultured human retinal endothelial cells (HRECs) subjected to BMP2 (50ng/ml) for up to 24 h. Also, effect of high glucose (HG, 30mM D-glucose) on the expression of BMP2 and its downstream genes was examined in HRECs. H2-DCF is a fluorogenic dye that measures the levels of cellular reactive oxygen species (ROS) was used to measure the pro-oxidative effect of BMP2. Moreover, we evaluated the effect of inhibiting p38 and VEGF signaling on BMP2-induced HRECs barrier dysfunction by measuring the trans-endothelial cell electrical resistance (TER) using electric cell-substrate impedance sensing (ECIS). We also tested the effect of HG on the integrity of HRECs barrier in the presence or absence of inhibitors of BMP2 signaling. Our data reveals that BMP2 and high glucose upregulates BMP components of the BMP signaling pathway (SMAD effectors, BMP receptors, and TGFβ ligand itself) and induces phosphorylation of p38 MAPK and NFκB with nuclear translocation of NFκB. Inhibition of p38 or NFκB attenuated BMP2-induced VEGF expression and barrier dysfunction in HRECs. Also, inhibition of VEGFR2 attenuated BMP2-induced barrier dysfunction. Moreover, BMP2 induces generation of ROS and endothelial nitric oxide synthase (eNOS) expression and activity in HRECs. Finally, HG upregulated BMP2 and its downstream genes (SMAD, BMP4, ALKs, and TGF-β) in HRECs and BMP2 inhibitors attenuated HG-induced HRECs barrier dysfunction. Our results suggest that in addition to the regular canonical SMAD signaling BMP2 induces non-canonical inflammatory pathway in HRECs via activation of p38/NFκB pathway that causes the upregulation of VEGF and the disruption of HRECs. Inhibition of BMP2 signaling is a potential therapeutic intervention to preserve endothelial cell barrier function in DR.
登录
查看更多内容
DOI：
10.1002/jor.22277
发表时间：
2013-04
期刊：
Journal of orthopaedic research : official publication of the Orthopaedic Research Society
影响因子：
--
作者：
Mattar T;Friedrich PF;Bishop AT
通讯作者：
Bishop AT
影响因子：
5.4
作者：
Hu J;Li T;Du S;Chen Y;Wang S;Xiong F;Wu Q
通讯作者：
Wu Q
影响因子：
7.5
作者：
Feng, Peng-Cheng;Ke, Xing-Fei;Wu, Jian-Bing
通讯作者：
Wu, Jian-Bing
影响因子：
6.5
作者：
Ibrahim, Ahmed S.;Elshafey, Sally;Al-Shabrawey, Mohamed
通讯作者：
Al-Shabrawey, Mohamed
DOI：
10.1016/j.bbalip.2014.12.017
发表时间：
2015-03
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
Ibrahim AS;Tawfik AM;Hussein KA;Elshafey S;Markand S;Rizk N;Duh EJ;Smith SB;Al-Shabrawey M
通讯作者：
Al-Shabrawey M