Department of Cardiology, Affiliated Hospital of Jiangsu University, Zhenjiang, Jiangsu 212013, P.R. China.
Department of Laboratory Medicine, Changzheng Hospital, Second Military Medical University, Shanghai 200003, P.R. China.
Mol Med Rep. 2021 Feb;23(2). doi: 10.3892/mmr.2020.11787. Epub 2020 Dec 23.
Patients with antiphospholipid syndrome have been identified to have higher incidence rates of atherosclerosis (AS) due to the elevated levels of anti‑β2‑glycoprotein I (β2GPI) antibody (Ab). Our previous studies revealed that the anti‑β2GPI Ab formed a stable oxidized low‑density lipoprotein (oxLDL)/β2GPI/anti‑β2GPI Ab complex, which accelerated AS development by promoting the accumulation of lipids in macrophages and vascular smooth muscle cell. However, the effects of the complex on endothelial cells, which drive the initiation and development of AS, remain unknown. Thus, the present study aimed to determine the proinflammatory roles of the oxLDL/β2GPI/anti‑β2GPI Ab complex in human umbilical vein endothelial cells (HUVECs) in an attempt to determine the underlying mechanism. Reverse transcription‑quantitative PCR, enzymy‑linked immunosorbent assay, western blotting and immunofluorescence staining were performed to detect the expressions of inflammation related factors and adhesion molecules. Monocyte‑binding assay was used to investigate the effects of oxLDL/β2GPI/anti‑β2GPI Ab complex on monocyte adhesion to endothelial cells. The results demonstrated that the oxLDL/β2GPI/anti‑β2GPI Ab complex upregulated the expression of Toll‑like receptor (TLR)4 and the levels of NF‑κB phosphorylation in HUVECs, and subsequently enhanced the expression levels of inflammatory cytokines, including TNF‑α, IL‑1β and IL‑6, as well as those of adhesion molecules, such as intercellular adhesion molecule 1 and vascular adhesion molecule 1. In addition, the complex facilitated the recruitment of monocytes by promoting the secretion of monocyte chemotactic protein 1 in HUVECs. Notably, the described effects of the oxLDL/β2GPI/anti‑β2GPI Ab complex in HUVECs were abolished by either TLR4 or NF‑κB blockade. In conclusion, these findings suggested that the oxLDL/β2GPI/anti‑β2GPI Ab complex may induce a hyper‑inflammatory state in endothelial cells by promoting the secretion of proinflammatory cytokines and monocyte recruitment, which was discovered to be largely dependent on the TLR4/NK‑κB signaling pathway.
患有抗磷脂综合征的患者由于抗β2-糖蛋白 I (β2GPI) 抗体 (Ab) 水平升高,其动脉粥样硬化 (AS) 的发病率更高。我们之前的研究表明,抗β2GPI Ab 形成了稳定的氧化型低密度脂蛋白 (oxLDL)/β2GPI/抗β2GPI Ab 复合物,通过促进巨噬细胞和血管平滑肌细胞中脂质的积累来加速 AS 的发展。然而,该复合物对内皮细胞的影响,内皮细胞驱动 AS 的起始和发展,仍不清楚。因此,本研究旨在确定 oxLDL/β2GPI/抗β2GPI Ab 复合物在人脐静脉内皮细胞 (HUVEC) 中的促炎作用,试图确定其潜在机制。采用逆转录-定量 PCR、酶联免疫吸附试验、western blot 和免疫荧光染色检测炎症相关因子和黏附分子的表达。采用单核细胞结合试验研究 oxLDL/β2GPI/抗β2GPI Ab 复合物对单核细胞与内皮细胞黏附的影响。结果表明,oxLDL/β2GPI/抗β2GPI Ab 复合物上调了 HUVEC 中 Toll 样受体 4 (TLR4) 和核因子-κB 磷酸化水平,进而增强了促炎细胞因子 TNF-α、IL-1β 和 IL-6 以及黏附分子如细胞间黏附分子 1 和血管细胞黏附分子 1 的表达水平。此外,该复合物通过促进单核细胞趋化蛋白 1 在 HUVEC 中的分泌,促进单核细胞的募集。值得注意的是,TLR4 或 NF-κB 阻断可消除 oxLDL/β2GPI/抗β2GPI Ab 复合物在 HUVECs 中的上述作用。综上所述,这些发现表明 oxLDL/β2GPI/抗β2GPI Ab 复合物可能通过促进促炎细胞因子的分泌和单核细胞的募集诱导内皮细胞发生过度炎症状态,而这主要依赖于 TLR4/NF-κB 信号通路。