LncRNA HOTAIR/miR-9-5p/FOXP1 axis modulates cerebral ischemia-reperfusion injury via NLRP3 inflammasome activation

LncRNA PRR34-AS1 Promoted the Neuroinflammation of Ischemic Stroke via Regulating miR-532-3p

Exosomal miR-451 Inhibits Ferroptosis by Targeting the PHD3/p53 Signaling Axis to Protect against Ischemic Stroke.

BACKGROUND Ischemic stroke causes irreversible neuronal injury. This study investigates whether exosomal miR-451 protects against stroke by inhibiting the PHD3/p53 pathway and attenuating ferroptosis in hippocampal neurons. METHODS An in vitro blood-brain barrier (BBB) model was established using co-cultured mouse hippocampal neurons (HT-22), astrocytes (C8-D1A), and brain microvascular endothelial cells (bEnd.3). Cerebral ischemia-reperfusion injury was mimicked by oxygen-glucose deprivation/reoxygenation (OGD/R). Barrier integrity was validated by fluorescein sodium permeability assay, 4-h liquid level difference assay, and immunofluorescence of tight junction protein ZO-1. Engineered exosomes loaded with miR-451 were characterized by nanoparticle size analysis and electron microscopy. Following exosome treatment, ferroptosis-related markers were assessed by qRT-PCR and immunofluorescence. A PHD3 overexpression plasmid was constructed to validate the involvement of the PHD3/p53 pathway, and bioinformatics analysis identified potential downstream targets of miR-451, including YTHDF2. RESULTS miR-451-loaded exosomes increased HT-22 viability and attenuated OGD/R-induced ferroptosis. Mechanistically, miR-451 downregulated PHD3 expression, reducing p53 levels. Bioinformatics analysis revealed a negative correlation between miR-451-targeted PHD3 and YTHDF2. YTHDF2 overexpression modulated ferroptosis markers, suggesting its regulatory role in ferroptosis. DISCUSSION These findings demonstrate that miR-451 confers neuroprotection via the PHD3/p53 axis to suppress ferroptosis, with YTHDF2 serving as an additional effector. The results underscore the therapeutic promise of exosome-delivered miR-451 for ischemic stroke treatment. CONCLUSION miR-451 protects against OGD/R-induced neuronal ferroptosis by targeting the PHD3/p53 axis, highlighting exosomal miR-451 as a potential therapeutic candidate for ischemic stroke.

Neuroprotective Effects of lncRNA EPB41L4A-AS1 Against Ischemic Stroke Injury Through Regulating Microglial State Transition and Ferroptosis via miR-214-3p/GPX4 Axis

miR-92a Agomir protects against ischemic stroke by suppressing oxidative stress, inflammation, excitotoxicity, and p53-mediated apoptosis.

Various microRNAs (miRs) have long served as therapeutic targets for ischemic stroke, so we aimed to assess whether miR-92a-3p protects against cerebral ischemia and identify downstream targets in an animal model. For this purpose, rats were randomly divided into sham, middle cerebral artery occlusion (MCAO), miR-92a Agomir+MCAO, and miR-92a Antagomir+MCAO groups. Twenty-four hours before MCAO surgery, rno-miR-92a-3p Agomir or Antagomir (100µM) were injected into the right striatum of rats. Neurological deficit, infarction, blood-brain barrier (BBB), cerebral edema, oxidative stress and inflammation-related factors mRNA expression (GCH1, GRIN1, NOS1, and NOX1), and the activities of antioxidant enzymes (SOD and CAT), and GSSG in the piriform cortex-amygdala, striatum, and cortex areas were evaluated. Immunohistochemistry was used to count positive cells of TNF-α and p53, and H&E staining served to examine histological alterations. Significant reductions in neurological deficit, infarction volume, edema, and BBB permeability were observed in the miR-92a Agomir+MCAO group compared to the MCAO group. miR-92a Agomir significantly down-regulated the expression of GCH1, GRIN1, NOS1, NOX1, TNF-α, and p53. The activities of antioxidant enzymes were elevated following administration of miR-92a Agomir. Moreover, miR-92a Agomir could attenuate the GSSG levels. While the preconditioning paradigm limits direct clinical translation, these mechanistic insights provide a compelling rationale for evaluating delayed administration strategies in future translational studies.

Protocatechualdehyde ameliorates OGD/R-induced endothelial injury via the ROS/miR-29b-3p/SIRT1 regulatory axis

Introduction MicroRNA-29b-3p (miR-29b-3p) plays a critical role in regulating endothelial apoptosis and autophagy. We previously demonstrated that protocatechualdehyde (PCA), a water-soluble phenolic acid derived from Salvia miltiorrhiza Bge., protects human umbilical vein endothelial cells (HUVECs) against oxygen-glucose deprivation/reoxygenation (OGD/R)-induced injury. However, whether miR-29b-3p mediates this protective effect remains unclear. Methods Using the OGD/R in vitro model of ischemia-reperfusion injury, we investigated the molecular mechanisms underlying ischemic damage at the cellular level. RT-qPCR was used to measure miR-29b-3p expression, Western blotting to assess autophagy- and apoptosis-related proteins, and MDC staining and flow cytometry to evaluate autophagy and apoptosis, respectively. Results The results showed that miR-29b-3p was downregulated in OGD/R-treated HUVECs, an effect that was reversed by PCA and the ROS inhibitor N-acetylcysteine (NAC). The miR-29b-3p mimic enhanced the promoting effects of PCA and NAC on miR-29b-3p expression, whereas miR-29b-3p inhibitor reversed these effects. Similarly, PCA-induced autophagy and inhibition of apoptosis were enhanced by the miR-29b-3p mimic and reversed by the inhibitor. To determine whether SIRT1 acts downstream of miR-29b-3p, we used EX527 (a SIRT1 inhibitor) and found that it attenuated the PCA-mediated upregulation of miR-29b-3p. The miR-29b-3p mimic enhanced PCA-induced SIRT1 activity and protein expression, while the inhibitor had the opposite effect. Additional experiments revealed that NAC suppressed ROS by upregulating miR-29b-3p, leading to increased SIRT1 expression and reduced apoptosis. Disscussion Collectively, these findings demonstrate that PCA ameliorates OGD/R-induced endothelial injury by promoting autophagy and inhibiting apoptosis via the ROS/miR-29b-3p/SIRT1 regulatory axis.

The LncRNA CRNDE/miR-152-3p/ROCK1 axis aggravates neurological deficits and neuroinflammation in intracerebral hemorrhage.

OBJECTIVE Intracerebral hemorrhage (ICH) is a fatal cerebrovascular disease. This study explored the intrinsic relationship between ICH and lncRNA CRNDE. METHODS Injecting the lentiviral interference vector (sh-CRNDE) into the ICH mice (induced by collagenase). Subsequently, the neurological deficits were evaluated through Garcia scoring, the corner turn test, and the water maze test. The levels of CRNDE and inflammatory factors in the brain tissue were detected by RT-qPCR and ELISA, respectively. In vitro experiments, microglia and neuronal cells were respectively cultured with hemoglobin and Hemin. Subsequently, the level of M1 polarization markers was detected by RT-qPCR. In neuronal cells, the proliferation activity was evaluated by CCK-8. The apoptosis level was comprehensively assessed based on the results of flow cytometry and the level of LDH. Finally, the effects of inhibiting the CRNDE/miR-152-3p axis on ROCK1 and the phenotype of ICH were re-verified in animal models and cell models. RESULTS In ICH mice, inhibition of CRNDE prevented the neurological deficits and neuroinflammation. Blocking CRNDE also significantly alleviated the M1 polarization of microglia and the apoptosis of neuronal cells. CRNDE sponges miR-152-3p through the ceRNA mechanism and regulates ROCK1. Inhibition of miR-152-3p or overexpression of ROCK1 weakens the inhibitory effect of sh-CRNDE on neurological deficits, M1 polarization, and the apoptosis of neurons. Moreover, in cell models, inhibition of ROCK1 showed the same protective effect as CRNDE downregulation. CONCLUSION In ICH, CRNDE upregulates ROCK1 by adsorbing miR-152-3p. This regulatory axis exacerbates neuroinflammation and neurofunctional disorders.