Experimental Study on the Regulation of Hypoxia/Reoxygenation-Induced Angiogenesis in Human Brain Microvascular Endothelial Cells by LncRNA-MEG3 via the miRNA-141-3p/Notch2 Axis
LIU Guangsheng
JIANG Yehan
ZHONG Juanli
PENG Luoquan
ZOU Ao
LIU Feng
Abstract：Objective An experimental study was conducted on the regulation of hypoxia/reoxygenation-induced angiogenesis in human brain microvascular endothelial cells by long non-coding RNA(LncRNA)-maternally expressed gene 3(MEG3)through the microRNA-141-3p/Notch homolog 2(Notch2)axis,to explore whether MEG3 affects the angiogenic capacity of hypoxia/reoxygenation human brain microvascular endothelial cells through the miRNA-141-3p/Notch2 axis.Methods An oxygen-glucose deprivation/reoxygenation(OGD/R)model was established to simulate the in vitro environment of ischemic stroke.Cell viability was detected by the Cell Counting Kit-8(CCK-8)assay,apoptosis was analyzed by flow cytometry,and the expression levels of genes and proteins were detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and Western blot,respectively.Cell migration and angiogenic ability were evaluated by Transwell and tube formation assays,respectively.The interaction between MEG3,poly-pyrimidine tract-binding protein 1(PTBP1),and Notch2 was analyzed by techniques such as RNA immunoprecipitation(RIP),RNA pull-down,and fluorescence in situ hybridization(FISH).In addition,a mouse middle cerebral artery occlusion(MCAO)model was established for in vivo validation.Results OGD/R treatment significantly upregulated the expression level of MEG3 in HBMECs,accompanied by decreased cell viability,increased apoptosis,and weakened migration and angiogenic ability.Knockdown of MEG3 reversed the above OGD/R-induced cell damage.Mechanistically,MEG3 was mainly located in the cytoplasm and directly bound to RNA-binding protein PTBP1,further regulating the stability of Notch2 messenger RNA(mRNA).Under OGD/R conditions,MEG3 enhanced Notch2 expression through PTBP1,thereby inhibiting the migration and angiogenesis of HBMECs.In vivo experiments further confirmed that silencing MEG3 significantly reduced the infarct area,improved neurological function scores,and reduced apoptosis.Conclusion MEG3 enhances the mRNA stability of Notch2 by binding to PTBP1,thereby inhibiting the migration and angiogenesis of HBMECs and promoting the development of ischemic stroke.Targeting the MEG3/PTBP1/Notch2 axis may provide a new potential target for the treatment of ischemic stroke.
Keywords：Ischemic strokeMaternally expressed gene 3Polypyrimidine tract binding protein 1Notch homolog 2MigrationAngiogenesis
Publication Date：2025-11-15
Online Publishing Date：2026-01-21(First online date of this platform, not the publication date of the document)
Pages：6( 1-6 )