MicroRNA‐212 promotes the recovery function and vascular regeneration of endothelial progenitor cells in mice with ischemic stroke through inactivation of the notch signaling pathway via downregulating MMP9 expression

Abstract

Ischemic stroke is a refractory disease caused by cerebral ischemic injury, which results in brain dysfunction. This study intends to investigate the effects of microRNA-212 (miR-212) on the recovery function and vascular regeneration of endothelial progenitor cells (EPCs) by inactivation of the Notch signaling pathway by binding to matrix metallopeptidase 9 (MMP9) in mice with ischemic stroke. According to the results of database retrieval systems and data analysis, MMP9 was predicted as a gene related to ischemic stroke and miR-212 is a potential regulating mRNA of MMP9. All 72 healthy adult C57BL6 mice were selected for middle cerebral artery occlusion (MCAO) establishment. Cerebral infarction was observed under triphenyltetrazolium chloride staining. A series of inhibitors, activators, and siRNAs were introduced to the verified regulatory functions for miR-212 governing MMP9 in ischemic stroke. Cell proliferation was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and tube-forming ability by tubule formation test. Reverse transcription quantitative polymerase chain reaction and Western blot analysis were used to detect the expressions of miR-212, MMP9, Hes-1, and Notch-1. The corresponding results demonstrated that the area of cerebral infarction and the number of neuronal necrosis increased in the MCAO group in contrast to the sham group. Meanwhile, upregulation of miR-212 or downregulation of MMP9 decreases the expressions of MMP9, Hes-1 Notch-1, increases cell proliferation and tube-forming ability and improves the pathological conditions of EPCs. Our study suggests that miR-212 promotes recovery function and vascular regeneration of EPCs through negative regulation of the Notch signaling pathway via downregulating expression of MMP9, thus provides a clinical theoretical basis for ischemic stroke therapy. K E Y W O R D S endothelial progenitor cells (EPCs), ischemic stroke, metallopeptidase 9 (MMP9), microRNA-212, notch pathway, vascular regeneration J Cell Physiol. 2019;234:7090-7103. wileyonlinelibrary.com/journal/jcp 7090 |

miR-212-5p attenuates ferroptotic neuronal death after traumatic brain injury by targeting Ptgs2

“…Previous studies implied that miR-212 is widely involved in the neuronal system [19]. It was related to the regeneration of endothelial progenitor cells [40], integrity of the blood brain barrier [23], dendritic morphology [41] and oligodendrocyte differentiation [42]. It has also been implicated in addiction-related and mental diseases [43][44][45].…”

Targeting neutrophils as a novel therapeutic strategy after stroke

“…Besides, Mir-212 is a potential regulating mRNA of MMP9, which can promote recovery function and vascular regeneration of EPCs via downregulating MMP9 expression in ischemic stroke. 108 Furthermore, icariside II (ICS II) has been found to significantly ameliorate I/R-induced BBB disruption and neuronal apoptosis in MCAO rats by regulating the MMP-9/ TIMP1 balance. 109 A recent study investigated a therapeutic strategy for stroke: substituting stroke mouse blood with whole blood obtained from naı¨ve, healthy donor mice, called blood replacement (BR) therapy.…”

Adding a “Notch” to Cardiovascular Disease Therapeutics: A MicroRNA-Based Approach

“…The miR-212/132 family inhibits FoxO3, an antihypertrophic and pro-autophagic transcription factor (Ucar et al, 2012) and an activator of Notch signaling (Gopinath et al, 2014). Notch inhibition by miR-212 has also been observed in mice with ischemic stroke, in which miR-212 promotes endothelial progenitor cells proliferation and tube formation through the inhibition of the Notch signaling (Hu and Dong, 2019). Similarly, miR-375 is increased in patients with HF (Akat et al, 2014) and is associated with cardiac hypertrophy and MI in rodent models (Feng et al, 2014;Garikipati et al, 2015).…”