The expression of cerebrospinal fluid exosomal miR-630 plays an important role in the dysfunction of endothelial cells after subarachnoid hemorrhage

Abstract

The purpose of this study was to evaluate the relationship of brain microvascular endothelial cell (BMECs) function and the exosomal miR-630 expression after subarachnoid hemorrhage (SAH). We evaluated the effects of blood cerebrospinal fluid (BCSF) on proliferation of BMECs by MTT at 0, 1, 3, 7 and 12 days and performed cell cycle analysis after BCSF treatment for 48 h. The expression of endothelial adhesion molecules (ICAM-1, VCAM-1 and ZO-1) were detected by qRT-PCR and immunofluorescent staining after BCSF treatment. NO produced by BMECs was also evaluated by Griess assay. The expression of exosomal miR-630 was analyzed by qRT-PCR in BCSF treated cell cultu normal cell culture medium andre medium. We further compared the exosomal miR-630 of clinical patients between aSAH and normal hydrocephalus. The adhesion molecules expression was further detected after co-incubation with exosomes transfected by miR-630 mimics. We found that BCSF significantly reduced the cell vitality in a time-dependent manner ( p < 0.05) and the growth inhibition ratio reached 78.34 ± 9.22% on the 12th day. BCSF induced cell cycle arrest in G0/G1 phase in BMECs ( p < 0.01). The expression of ICAM-1, VCAM-1, ZO-1 and the NO produced by BMECs were markedly reduced following incubation with BCSF. Then we demonstrated that the expression of exosomal miR-630 was markedly reduced in the BCSF treated BMECs and the same phenomenon occurred in aSAH patients compared with normal hydrocephalus. The expression of ICAM-1, VCAM-1 and ZO-1 were then increased in BMECs cocultured with exosomes transfected by miR-630 mimics. In conclusion, the low expression of exosomal miR-630 in CSF was closely related to endothelial function in BCSF endothelial cell injury model and clinical patients.

Systemic exosomal miR-193b-3p delivery attenuates neuroinflammation in early brain injury after subarachnoid hemorrhage in mice

“…Previous studies demonstrated that miRNAs were involved in numerous physiological/pathological processes and were particularly appealing diagnostic and therapeutic tools for various diseases, including stroke, Parkinson's disease, traumatic brain injury (TBI), and Alzheimer's disease. Despite the fact that expression levels of miRNAs have measured in cerebrospinal fluid and in circulation, there are few studies investigating changes of miRNAs after SAH [12][13][14]. The blood-brain barrier (BBB) has been proven to be a major obstacle for delivery of drugs to the brain.…”

Advances in Analytical Technologies for Extracellular Vesicles

“…Exosomes and other EVs have been confirmed to be present in almost all the body fluids since first discovery, including blood, − pleural effusions, , aqueous humor, , breast milk, , ascites, amniotic fluid, semen, saliva, , nasal secretions, cerebrospinal fluid, , bronchoalveolar lavage, synovial fluid, , bile, urine, , cell culture-conditioned media, , etc . These EVs containing proteins and NAs provide a rich resource to identify biomarkers for disease diagnosis.…”

The blood–brain barrier and the neurovascular unit in subarachnoid hemorrhage: molecular events and potential treatments

“…MicroRNA (miR)-630 may also play a role in the expression of adhesion molecules and TJ proteins. Low miR-630 expression was found in endothelial cells treated with arterial blood, indicating a crucial role for exosomal miR-630 in maintaining BBB integrity after SAH [ 211 ]. Periostin, one of the matricellular proteins, activates the MAPK signaling pathway through integrins and modulates downstream pathways such as MMP-9 after SAH.…”