Tian Dan, Qin Qing, Li Mingfei, Li Xiaoyu, Xu Qing, Lv Qianzhou
Department of Pharmacy, Zhongshan Hospital, Fudan University, Shanghai, China.
Department of Cardiology, Zhongshan Hospital, Fudan University, Shanghai Institute of Cardiovascular Disease, Shanghai, China.
Front Pharmacol. 2021 Jan 8;11:614760. doi: 10.3389/fphar.2020.614760. eCollection 2020.
Hyperhomocysteinemia is a well-recognized independent risk factor for cardiovascular disease. To date, the mechanism of pathological plasma homocysteine (Hcy) level elevation remains to be elucidated. We aimed to investigate the levels of progranulin (PGRN), Eph-receptor tyrosine kinase-type A2 (EphA2), vascular cell adhesion molecule-1 (VCAM-1), and Hcy in patients with arteriosclerosis and investigate their functions in Hcy-injured human umbilical vein endothelial cells (HUVECs). EphA2 knockdown was induced in HUVECs by shRNA lentivirus infection with EphA2-RNAi, and bulk RNA-seq assay was performed. Then we investigated the mechanism underlying the effect of recombinant human PGRN (rhPGRN) combined with shRNA interference of EphA2 on cell proliferation, migration, and angiogenesis in Hcy-injured HUVECs. Results showed that serum EphA2, VCAM-1, and Hcy levels in acute coronary syndrome patients were significantly higher than those in chronic coronary syndrome patients ( = 0.000; = 0.000; = 0.033, respectively). , we demonstrated that knockdown of EphA2 significantly impaired cell adhesion and inhibited HUVECs migration and angiogenesis ( < 0.001), which was associated with reduction in VCAM1 and VE-cadherin ( < 0.05). Hcy modulated the expression of PGRN and EphA2 in a time-and dose-dependent manner. However, rhPGRN ameliorated the Hcy-induced reduction in cell viability and migration ( < 0.05). Mechanistically, we found that PGRN/EphA2 and its downstream AKT/NF-κB signaling might be the primary signal transduction pathways underlying Hcy-induced injury. The present study illustrated that PGRN plays a previously unrecognized role in Hcy-induced endothelial injury, which is achieved through its interaction with EphA2 signaling, implying a promising therapeutic target for cardiovascular disease.
高同型半胱氨酸血症是一种公认的心血管疾病独立危险因素。迄今为止，病理性血浆同型半胱氨酸（Hcy）水平升高的机制仍有待阐明。我们旨在研究动脉硬化患者中颗粒蛋白前体（PGRN）、Eph受体酪氨酸激酶A2型（EphA2）、血管细胞黏附分子-1（VCAM-1）和Hcy的水平，并研究它们在Hcy损伤的人脐静脉内皮细胞（HUVECs）中的作用。通过用EphA2-RNAi的shRNA慢病毒感染在HUVECs中诱导EphA2敲低，并进行批量RNA测序分析。然后我们研究了重组人PGRN（rhPGRN）联合EphA2的shRNA干扰对Hcy损伤的HUVECs细胞增殖、迁移和血管生成影响的潜在机制。结果显示，急性冠状动脉综合征患者的血清EphA2、VCAM-1和Hcy水平显著高于慢性冠状动脉综合征患者（分别为P = 0.000；P = 0.000；P = 0.033）。此外，我们证明EphA2敲低显著损害细胞黏附并抑制HUVECs迁移和血管生成（P < 0.001），这与VCAM1和血管内皮钙黏蛋白的减少有关（P < 0.05）。Hcy以时间和剂量依赖性方式调节PGRN和EphA2的表达。然而，rhPGRN改善了Hcy诱导的细胞活力和迁移降低（P < 0.05）。从机制上讲，我们发现PGRN/EphA2及其下游的AKT/NF-κB信号可能是Hcy诱导损伤的主要信号转导途径。本研究表明，PGRN在Hcy诱导的内皮损伤中发挥了以前未被认识的作用，这是通过其与EphA2信号的相互作用实现的，这意味着它是心血管疾病一个有前景的治疗靶点。
