Department of Neurosurgery, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510630, China.
Department of Radiology, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510630, China.
Cytokine. 2022 Sep;157:155973. doi: 10.1016/j.cyto.2022.155973. Epub 2022 Jul 27.
Intracranial aneurysm (IA) is cerebrovascular disorder which refers to local vessel wall damage to intracranial arteries, forming abnormal bulge. Both endothelial cells (ECs) and vascular smooth muscle cells (VSMCs) are closely associated with IA formation and rupture. Inflammatory SMCs (iSMCs) were reported to induce EC dysfunction and result in IA progression. Phoenixin-14 (PNX-14) is a recently discovered brain peptide with pleiotropic roles, which participates in reproduction, cardio protection, lipid deposition and blood glucose metabolism. PNX-14 was previously reported to protect brain endothelial cells against oxygen-glucose deprivation/reoxygenation (OGD/R)-induced cell injury. Therefore, our study was designed to investigate the influence of PNX-14 on iSMCs-induced endothelial dysfunction.
Inflammation in SMCs was induced by cyclic mechanical stretch. Human umbilical vein endothelial cells (HUVECs) were exposed to SMC- or iSMC-conditioned medium and then treated with 100 nM PNX-14 for 24 h. The levels of proinflammatory cytokines (IL-1β, IL-6 and TNF-α) in cell supernatants were analyzed by ELISA. Cell viability, apoptosis, angiogenesis and migration were subjected to CCK-8 assay, flow cytometry analysis, tube formation assay and Transwell migration assay. The protein levels of proinflammatory cytokines and apoptosis markers (Bcl-2 and Bax) were evaluated by western blotting.
Cyclic mechanical stretch upregulated IL-1β, IL-6 and TNF-α levels in SMCs. Treatment with SMC- or iSMC-conditioned medium HUVECs inhibited cell viability, angiogenesis and migration and induced apoptosis in HUVECs. iSMC-conditioned medium has more significant effects on cell functions. However, the influence of SMC- or iSMC-conditioned medium treatment on HUVEC biological functions were reversed by PNX-14 treatment. PNX-14 exerts no significant influence on the biological functions of HUVECs treated with SMC medium.
PNX-14 alleviates iSMCs-induced endothelial cell dysfunction in vitro.
颅内动脉瘤（IA）是一种脑血管疾病，指颅内动脉的局部血管壁损伤，形成异常膨出。内皮细胞（ECs）和血管平滑肌细胞（VSMCs）都与 IA 的形成和破裂密切相关。炎性平滑肌细胞（iSMCs）被报道可诱导 EC 功能障碍，从而导致 IA 的进展。凤凰素-14（PNX-14）是一种新发现的具有多种作用的脑肽，参与生殖、心脏保护、脂质沉积和血糖代谢。先前有研究报道，PNX-14 可保护脑内皮细胞免受氧葡萄糖剥夺/再氧合（OGD/R）诱导的细胞损伤。因此，本研究旨在探讨 PNX-14 对 iSMCs 诱导的内皮功能障碍的影响。
通过循环机械拉伸诱导 SMC 炎症。将人脐静脉内皮细胞（HUVECs）暴露于 SMC 或 iSMC 条件培养基中，然后用 100 nM PNX-14 处理 24 小时。通过 ELISA 分析细胞上清液中促炎细胞因子（IL-1β、IL-6 和 TNF-α）的水平。通过 CCK-8 测定、流式细胞术分析、管形成测定和 Transwell 迁移测定来检测细胞活力、凋亡、血管生成和迁移。通过 Western blot 评估促炎细胞因子和凋亡标志物（Bcl-2 和 Bax）的蛋白水平。
循环机械拉伸可上调 SMC 中 IL-1β、IL-6 和 TNF-α 的水平。用 SMC 或 iSMC 条件培养基处理 HUVECs 可抑制细胞活力、血管生成和迁移，并诱导 HUVECs 凋亡。iSMC 条件培养基对细胞功能的影响更为显著。然而，PNX-14 处理可逆转 SMC 或 iSMC 条件培养基处理对 HUVEC 生物学功能的影响。PNX-14 对用 SMC 培养基处理的 HUVECs 的生物学功能没有显著影响。
PNX-14 可减轻体外 iSMCs 诱导的内皮细胞功能障碍。
