Medical School, Kunming University of Science and Technology, Kunming, 650500, PR China; The Department of Cardiovascular Medicine, State Key Laboratory of Medical Genomics, Shanghai Key Laboratory of Hypertension, Ruijin Hospital, Shanghai Institute of Hypertension, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, PR China.
The Department of Cardiovascular Medicine, State Key Laboratory of Medical Genomics, Shanghai Key Laboratory of Hypertension, Ruijin Hospital, Shanghai Institute of Hypertension, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, PR China.
Biochem Biophys Res Commun. 2023 Jul 23;666:76-82. doi: 10.1016/j.bbrc.2023.05.012. Epub 2023 May 3.
Vascular endothelial cell (VEC) apoptosis is the fundamental cause of pulmonary arterial hypertension. MicroRNA-31 (MiR-31) is a novel target for hypertension treatment. However, the role and mechanism of miR-31 in the apoptosis of VECs remain unclear. The purpose of this study is to determine whether miR-31 plays an important role in VEC apoptosis as well as the detailed mechanisms involved. We found that pro-inflammatory cytokines IL-17A and TNF-α were highly expressed in serum and aorta, and the expression of miR-31 was significantly increased in aortic intimal tissue from Angiotensin II (AngII)- induced hypertensive mice (WT-AngII) compared with control mice (WT-NC). In vitro, co-stimulation of VECs with IL-17A and TNF-α resulted in increased expression of miR-31 and VEC apoptosis. MiR-31 inhibition strikingly decreased TNF-α and IL-17A co-induced VEC apoptosis. Mechanistically, in IL-17A and TNF-α co-stimulated VECs (co-induced VECs), we found that the activation of the NF-κB signal effectively increased the expression of miR-31. Dual-luciferase reporter gene assay revealed that miR-31 directly targeted and inhibited the expression of the E2F transcription factor 6 (E2F6). The expression of E2F6 was decreased in Co-induced VECs. MiR-31 inhibition significantly alleviated the decreased expression of E2F6 in co-induced VECs. Consistent with the co-stimulated effect of IL-17A and TNF-α on VECs, transfection of siRNA E2F6 induced cell apoptosis without the stimulation of the above cytokines. In conclusion, TNF-α and IL-17A generated in the aortic vascular tissue and serum from Ang II-induced hypertensive mice could trigger VECs apoptosis by the miR-31/E2F6 axis. To sum up, our study suggests that the key factor between cytokine co-stimulation effect and VEC apoptosis was miR-31/E2F6 axis, which was mainly regulated by NF-қB signaling pathway. This gives us a new sight to treat hypertension-associated VR.
血管内皮细胞（VEC）凋亡是肺动脉高压的根本原因。微小 RNA-31（miR-31）是高血压治疗的新靶点。然而，miR-31 在 VEC 凋亡中的作用和机制尚不清楚。本研究旨在确定 miR-31 是否在 VEC 凋亡中发挥重要作用以及涉及的详细机制。我们发现，促炎细胞因子 IL-17A 和 TNF-α在血清和主动脉中高表达，血管紧张素 II（AngII）诱导的高血压小鼠（WT-AngII）主动脉内膜组织中 miR-31 的表达明显增加，而对照组小鼠（WT-NC）则没有。在体外，IL-17A 和 TNF-α共同刺激 VEC 导致 miR-31 表达增加和 VEC 凋亡。miR-31 抑制可显著减少 TNF-α 和 IL-17A 共同诱导的 VEC 凋亡。在 IL-17A 和 TNF-α 共同刺激的 VEC（共同诱导的 VEC）中，我们发现 NF-κB 信号的激活有效地增加了 miR-31 的表达。双荧光素酶报告基因检测显示，miR-31 可直接靶向并抑制 E2F 转录因子 6（E2F6）的表达。共同诱导的 VEC 中 E2F6 的表达减少。miR-31 抑制可显著减轻共同诱导的 VEC 中 E2F6 的表达降低。与 IL-17A 和 TNF-α 对 VEC 的共同刺激作用一致，转染 siRNA E2F6 可诱导细胞凋亡，而无需上述细胞因子的刺激。总之，在 Ang II 诱导的高血压小鼠的主动脉血管组织和血清中产生的 TNF-α 和 IL-17A 可以通过 miR-31/E2F6 轴触发 VEC 凋亡。综上所述，我们的研究表明，细胞因子共同刺激作用与 VEC 凋亡之间的关键因素是 miR-31/E2F6 轴，其主要受 NF-қB 信号通路调节。这为我们治疗高血压相关 VR 提供了新的视角。
