Pharmaceutical Department, Zhongshan Torch Development Zone Hospital, Zhongshan, Guangdong, China.
Department of Circulatory Medicine, Zhongshan Torch Development Zone Hospital, Zhongshan, Guangdong, China.
Neurosci Lett. 2022 Jan 23;770:136381. doi: 10.1016/j.neulet.2021.136381. Epub 2021 Dec 11.
The maintenance of human brain microvascular endothelial cell (HBMEC) function is crucial to improve the outcomes of ischemic stroke (IS). Emerging evidence shows that circular RNAs (circRNAs) are involved in IS progression. This study aimed to investigate the role of circRNA FUN14 domain containing 1 (circFUNDC1) in oxygen-glucose deprivation (OGD)-treated HBMECs.
The expression of circFUNDC1, miR-375 and phosphatase and tensin homolog (PTEN) mRNA was detected by quantitative real-time PCR (qPCR). Cell viability, apoptosis, migration and angiogenesis were determined by CCK-8 assay, flow cytometry assay, transwell assay and tube formation assay. The protein level of PTEN was detected by western blot. The relationship between miR-375 and circFUNDC1 or PTEN was confirmed by pull-down assay, dual-luciferase reporter assay and RIP assay. Exosomes were identified by transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA).
CircFUNDC1 expression was increased in peripheral blood of IS patients and OGD-treated HBMECs. CircFUNDC1 knockdown alleviated OGD-induced cell apoptosis and promoted OGD-blocked cell viability, migration and angiogenesis of HBMECs. MiR-375 was a target of circFUNDC1, and miR-375 restoration played similar effects with circFUNDC1 knockdown. The inhibition of miR-375 reversed the effects of circFUNDC1 knockdown. In addition, PTEN was a downstream target of miR-375, and PTEN overexpression abolished the effects of miR-375 restoration. The expression of circFUNDC1 was elevated in serum-derived exosomes of IS patients, and circFUNDC1 harbored diagnostic values.
CircFUNDC1 knockdown alleviates OGD-induced HBMECs injuries by inhibiting PTEN via enriching miR-375.
维持人脑血管内皮细胞(HBMEC)功能对于改善缺血性脑卒中(IS)的预后至关重要。新出现的证据表明,环状 RNA(circRNA)参与了 IS 的进展。本研究旨在探讨 FUN14 结构域包含 1(circFUNDC1)环状 RNA 在氧葡萄糖剥夺(OGD)处理的 HBMEC 中的作用。
通过实时定量 PCR(qPCR)检测 circFUNDC1、miR-375 和磷酸酶张力蛋白同源物(PTEN)mRNA 的表达。通过 CCK-8 测定法、流式细胞术测定法、Transwell 测定法和管形成测定法测定细胞活力、凋亡、迁移和血管生成。通过 Western blot 检测 PTEN 蛋白水平。通过下拉测定法、双荧光素酶报告基因测定法和 RIP 测定法证实 miR-375 与 circFUNDC1 或 PTEN 之间的关系。通过透射电子显微镜(TEM)和纳米颗粒跟踪分析(NTA)鉴定外泌体。
IS 患者外周血和 OGD 处理的 HBMEC 中 circFUNDC1 的表达增加。circFUNDC1 敲低减轻了 OGD 诱导的细胞凋亡,并促进了 OGD 阻断的 HBMEC 活力、迁移和血管生成。miR-375 是 circFUNDC1 的靶标,miR-375 恢复与 circFUNDC1 敲低具有相似的作用。miR-375 抑制逆转了 circFUNDC1 敲低的作用。此外,PTEN 是 miR-375 的下游靶标,PTEN 过表达消除了 miR-375 恢复的作用。IS 患者血清来源的外泌体中 circFUNDC1 的表达升高,并且 circFUNDC1 具有诊断价值。
circFUNDC1 敲低通过富集 miR-375 抑制 PTEN 减轻 OGD 诱导的 HBMEC 损伤。