1α,25-dihydroxyvitamin D3 inhibits tumor necrosis factor-α induced activation of human umbilical vein endothelial cells via NF-κB signaling pathway
10.3760/cma.j.issn.1000-6699.2019.06.010
- VernacularTitle:1α,25-二羟维生素D3通过NF-Κb信号通路抑制肿瘤坏死因子-α诱导的人脐静脉内皮细胞活化
- Author:
Yishan ZHOU
1
;
Qingyan ZHANG
;
Chunming JIANG
;
Yuan FENG
;
Jing LIU
;
Bo JIN
;
Nan LI
;
Miao ZHANG
Author Information
1. 南京大学医学院附属鼓楼医院肾内科 210008
- Keywords:
1α;
25-dihydroxyvitamin D3;
Endothelial activation;
NF-κB signaling pathway;
Adhesion molecule
- From:
Chinese Journal of Endocrinology and Metabolism
2019;35(6):499-505
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of 1α, 25-dihydroxyvitamin D3 [1α,25-(OH)2 D3] on tumor necrosis factor-α ( TNF-α) induced activation of human umbilical vein endothelial cells ( HUVECs ) . The mechanism involved in this process was also studied. Methods HUVECs were cultured and treated with TNF-α( 40 ng/ml), 1α,25-(OH)2D3(10-8 mol/L), and SN50 as indicated. Vascular cell adhesion molecule (VCAM) and E-selectin were used as markers of endothelial activation, which were detected by Western blotting and realtime PCR (RT-PCR). NF-κB signaling pathway was investigated to study the mechanism. Western blotting, RT-PCR, immunofluorescence assay, and chromatin immunoprecipitation ( ChIP ) method were used to evaluate the effects of 1α,25-( OH) 2 D3 on its early activation, nuclear transport, and binding to VCAM and E-selectin promoters. Results (1) Western blotting and RT-PCR showed that TNF-α could significantly up-regulate the expression of VCAM and E-selectin in HUVECs, which can be inhibited by specific NF-κB blocker SN50. 1α,25-( OH) 2 D3 down-regulated the expression of VCAM and E-selectin induced by TNF-α. ( 2 ) Western blotting showed that TNF-α induces I-κBαphosphorylation, thereby activating NF-κB p65 subunit. Immunofluorescence showed that 1α, 25-( OH ) 2 D3 significantly inhibited the nuclear translocation of NF-κB p65 subunit. ChIP analysis revealed that 1α,25-( OH) 2 D3 inhibited the binding of NF-κB p65 to VCAM and E-selectin promoters and thus affected gene expression. Conclusions TNF-αenhanced the expression of E-selectin and VCAM in HUVECs via NF-κB signaling pathway. 1α,25-( OH) 2 D3 may inhibit NF-κB early activation, nuclear transport and the binding of NF-κB p65 to VCAM and E-selectin promoters, thereby inhibiting TNF-α-induced endothelial cell activation.
