miR-140-5p regulates angiogenesis following ischemic stroke by targeting VEGFA

Abstract

MicroRNA (miRNA or miR) expression profiles are altered in tissues under hypoxic-ischemic conditions. The expression of miR‑140 is downregulated >2-fold following hypoxic-ischemic brain damage, however, its role in angiogenesis subsequent to cerebral ischemia is not fully understood. The present study aimed to investigate the role of miR-140-5p in angiogenesis and the molecular mechanism mediated by vascular endothelial growth factor A (VEGFA) in an in vitro model for brain ischemia. A rat middle cerebral artery occlusion (MCAO) model was constructed, and the results from reverse transcription-quantitative polymerase chain reaction and western blot analysis demonstrated that the expression levels of miR-140‑5p were significantly decreased, while the expression levels of VEGFA were significantly increased between 12 and 48 h in the rat cerebral following MCAO. Furthermore, human umbilical vein endothelial cells (HUVECs) were exposed to low oxygen conditions and it was demonstrated that hypoxia downregulated miR-140-5p and upregulated VEGFA expression levels. The miR-140-5p mimic was transfected into the normoxic and hypoxic HUVECs and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, Transwell migration and tube formation assays were performed. The results indicated that miR‑140‑5p inhibited angiogenesis by decreasing cell proliferation, migration and tube formation. Additionally, in human embryonic kidney 293 cells, results from the luciferase reporter assay revealed that miR‑140‑5p directly targeted the 3' untranslated region of VEGFA and that miR‑140‑5p regulated the protein expression of VEGFA. To further analyze this effect, a VEGFA‑pEGFP‑C1 plasmid was transfected into the normoxic and hypoxic HUVECs, and it was revealed that the inhibitory effect of miR‑140‑5p on angiogenesis was attenuated by the overexpression of VEGFA. In conclusion, to the best of our knowledge, the present study is the first to suggest that miR‑140‑5p exerts an inhibitory effect on angiogenesis in an in vitro model of ischemia, and this effect is achieved partially by targeting VEGFA. The present study provided a novel biomarker for the treatment of cerebral ischemia.

Retracted: MicroRNA‐140‐5p elevates cerebral protection of dexmedetomidine against hypoxic–ischaemic brain damage via the Wnt/β‐catenin signalling pathway

“…Mayorga et al 26 pointed out that miR‐145 expression could be used as a regulator of activity of β‐catenin in terms of hypoxia and ischaemia. Moreover, miR‐140‐5p could inhibit angiogenesis in a rat model of ischaemia by targeting VEGFA, which may be beneficial for cerebral ischaemia treatment 8. Additionally, it was demonstrated during our study that the mRNA and protein expressions of Wnt1, β‐catenin, TCF‐4 and E‐cadherin were increased in other groups as compared with the model group.…”

“…The abnormal expression of miR‐140 has been reported to be induced after HIBD. DEX has been proven to regulate HIBD 8, 15. In an attempt to elucidate a potential HIBD therapeutic target, an investigation was pursued into miR‐140‐5p and its effects on cerebral protection of DEX against HIBD by targeting Wnt1 through the Wnt/β‐catenin signalling pathway in neonatal rats.…”

“…At present, treatment options are confined to supportive care due to the complex mechanism and severity of the condition 7. In addition, previous studies have provided evidence suggesting that microRNAs (miRNAs) are aberrantly expressed in the tissues of rat models with ischaemic stroke, such as miR‐140 8. Thus, it is feasible to explore the highlighting the clinical research value of miRNAs and the unearthing the underlying mechanism behind HIBD in neonatal rats.…”

“…Furthermore, miR‐140 expression is notably decreased after HIBD. In addition, studies have revealed miR‐140‐5p plays an essential angiogenesis role in the ischaemia model via its regulation of VEGFA, which may be beneficial for cerebral ischaemia treatment 8. The combination of miRNAs reduction and Wnt/β‐catenin signalling pathway could promote cancer metastasis, carcinogenesis and even drug resistance 11.…”

Identification and Verification of Candidate Genes Regulating Neural Stem Cells Behavior Under Hypoxia

“…Subsequently, we performed a search in PubMed for these 14 miRNAs and found that 7 of them had been confirmed experimentally to target VEGFA, that is, miR-361 [40, 41], miR-140 [42], miR-15a [43], miR-15b [44], miR-205 [45], miR-206 [46], and miR-200c [47]. Therefore, we speculate that these miRNAs may target VEGFA and play a role in NSC behavior under hypoxia.…”