Department of Neurology, The Second Clinical Medical College of Jinan University (Shenzhen People's Hospital), No. 1017, Dongmen North Road, Shenzhen, 518020, Guangdong, China.
Department of Neurology, The Second Clinical Medical College of Jinan University (Shenzhen People's Hospital), No. 1017, Dongmen North Road, Shenzhen, 518020, Guangdong, China.
Mol Cell Probes. 2022 Aug;64:101830. doi: 10.1016/j.mcp.2022.101830. Epub 2022 May 27.
Cerebral ischemia is a serious acute disease with high mortality and disability rates. circRNAs are associated with cerebral ischemia inflammation and can be used as therapeutic targets. We aimed to investigate circNup188/miR-760-3p/Map3k8 role in inflammation during cerebral ischemia.
According to the sequencing results of previously published, signaling pathways and circRNAs related to inflammation were screened and looped for verification. In vitro OGD/R cell model was constructed to detect OGD/R effects on PC12 cell viability, apoptosis, inflammatory cytokines TNF-α, IL-1β and IL-6. qRT-PCR assessed circRNAs, circNup188 bound miRNAs, and four mRNAs associated with inflammation and brain diseases expressions. p65 and IkB-α and their phosphorylated proteins in NF-κB pathway and Map3k8 expressions were assessed by Western blot.
KEGG analysis revealed MAPK and NF-kappaB signaling pathways played a vital role in it, and were classic inflammatory pathways. circNup188, circU2, and circCnot2 were verified to be the loop structure. circNup188 might play an essential role in OGD/R cell model. Regulating circNup188 expression could affect OGD/R cells function. In addition, miR-760-3p might play a vital role in OGD/R cell model. Moreover, circNup188 regulated cerebral ischemia by sponging miR-760-3p. miR-760-3p might be involved in inflammation in OGD/R cell model by regulating Map3k8 to activate the NF-κB pathway.
circNup188 might up-regulate Map3k8 expression through sponging miR-760-3p and then activate NF-κB pathway, which was involved in cerebral ischemia development. This study provided a new target for cerebral ischemia treatment.
脑缺血是一种死亡率和致残率都很高的严重急性疾病。circRNAs 与脑缺血炎症有关，可作为治疗靶点。我们旨在研究 circNup188/miR-760-3p/Map3k8 在脑缺血炎症中的作用。
根据先前发表的测序结果，筛选并验证了与炎症相关的信号通路和 circRNAs。构建体外 OGD/R 细胞模型，检测 OGD/R 对 PC12 细胞活力、凋亡、炎症细胞因子 TNF-α、IL-1β 和 IL-6 的影响。qRT-PCR 评估 circRNAs、circNup188 结合的 miRNAs 以及与炎症和脑部疾病相关的四个 mRNAs 的表达。通过 Western blot 评估 NF-κB 通路中的 p65 和 IkB-α 及其磷酸化蛋白和 Map3k8 的表达。
KEGG 分析显示 MAPK 和 NF-κB 信号通路在其中发挥了重要作用，并且是经典的炎症通路。验证了 circNup188、circU2 和 circCnot2 是其环结构。circNup188 可能在 OGD/R 细胞模型中发挥重要作用。调节 circNup188 的表达会影响 OGD/R 细胞的功能。此外，miR-760-3p 可能在 OGD/R 细胞模型中发挥重要作用。此外，circNup188 通过海绵吸附 miR-760-3p 调节脑缺血。miR-760-3p 可能通过调节 Map3k8 激活 NF-κB 通路，参与 OGD/R 细胞模型中的炎症。
circNup188 可能通过海绵吸附 miR-760-3p 上调 Map3k8 表达，然后激活 NF-κB 通路，参与脑缺血的发生。本研究为脑缺血治疗提供了新的靶点。
