CircSEC11A knockdown 通过调控 miR-29a-3p/SEMA3A 轴减轻 OGD 诱导的人脑微血管内皮细胞（HBMECs）中的氧化应激和细胞凋亡，并促进细胞增殖和血管生成。
CircSEC11A knockdown alleviates oxidative stress and apoptosis and promotes cell proliferation and angiogenesis by regulating miR-29a-3p/SEMA3A axis in OGD-induced human brain microvascular endothelial cells (HBMECs).
机构信息
Department of Neurology, Wuhan Puren Hospital Affiliated to Wuhan University of Science and Technology, Wuhan, Hubei, China.
出版信息
Clin Hemorheol Microcirc. 2023;84(3):247-262. doi: 10.3233/CH-221689.
BACKGROUND
Circular RNA (circRNA) has been found to play an important role in the progression of many diseases, including ischemic stroke. However, the regulatory mechanism of circSEC11A in ischemic stroke progression need to further investigation.
METHODS
Human brain microvascular endothelial cells (HBMECs) were stimulated by oxygen glucose deprivation (OGD). CircSEC11A, SEC11A mRNA and miR (microRNA)-29a-3p were quantified by quantitative real-time PCR (qRT-PCR). SEMA3A, BAX and BCL2 protein level was quantified by western blot. Oxidative stress, cell proliferation, angiogenesis and apoptosis abilities were gauged by oxidative stress assay kit, 5-Ethynyl-2'-Deoxyuridine (EdU) staining, tube formation assay and flow cytometry assays, respectively. Direct relationship between miR-29a-3p and circSEC11A or SEMA3A was validated by dual-luciferase reporter assay, RIP assay and RNA pull-down assay.
RESULTS
CircSEC11A was upregulated in OGD-induced HBMECs. OGD promoted the oxidative stress and apoptosis and inhibited cell proliferation and angiogenesis, while circSEC11A knockdown relieved the effects. CircSEC11A functioned as the sponge for miR-29a-3p, and miR-29a-3p inhibitor reversed the effects of si-circSEC11A on OGD-induced HBMECs oxidative injuries. Moreover, SEMA3A served as the target gene of miR-29a-3p. MiR-29a-3p inhibition ameliorated OGD-induced HBMECs oxidative injuries, while SEMA3A overexpression rescued the impacts of miR-29a-3p mimic.
CONCLUSION
CircSEC11A promoted the malignant progression in OGD-induced HBMECs through the mediation of miR-29a-3p/SEMA3A axis. This study has provided the new insight into the underlying application of circSEC11A in cell model of ischemic stroke.
背景
环状 RNA（circRNA）已被发现在许多疾病的进展中发挥重要作用，包括缺血性中风。然而，circSEC11A 在缺血性中风进展中的调节机制仍需进一步研究。
方法
用氧葡萄糖剥夺（OGD）刺激人脑微血管内皮细胞（HBMEC）。用定量实时 PCR（qRT-PCR）定量检测 circSEC11A、SEC11A mRNA 和 miR（microRNA）-29a-3p。用 Western blot 定量检测 SEMA3A、BAX 和 BCL2 蛋白水平。用氧化应激检测试剂盒、5-乙炔基-2'-脱氧尿苷（EdU）染色、管形成测定和流式细胞术分别测定氧化应激、细胞增殖、血管生成和细胞凋亡能力。通过双荧光素酶报告基因检测、RIP 检测和 RNA 下拉实验验证 miR-29a-3p 与 circSEC11A 或 SEMA3A 之间的直接关系。
结果
在 OGD 诱导的 HBMECs 中，circSEC11A 上调。OGD 促进氧化应激和细胞凋亡，抑制细胞增殖和血管生成，而 circSEC11A 敲低则缓解了这些作用。circSEC11A 作为 miR-29a-3p 的海绵，miR-29a-3p 抑制剂逆转了 si-circSEC11A 对 OGD 诱导的 HBMECs 氧化损伤的作用。此外，SEMA3A 是 miR-29a-3p 的靶基因。miR-29a-3p 抑制可改善 OGD 诱导的 HBMECs 氧化损伤，而 SEMA3A 过表达可挽救 miR-29a-3p 模拟物的影响。
结论
circSEC11A 通过介导 miR-29a-3p/SEMA3A 轴促进 OGD 诱导的 HBMECs 中的恶性进展。本研究为 circSEC11A 在缺血性中风细胞模型中的潜在应用提供了新的见解。
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