Project description:hCMEC/D3 cells were treated with SiNPs for 24 h. It was discovered that SiNP_35 nm and SiNP_58 nm were both penetrated through the BBB via caveolin-mediated pathway, and both sizes induced the upregulation of the inflammation pathways.
Project description:In the current study, we aimed to investigate the gene expression pattern in hCMEC/D3 cells using mRNA microarray analysis and to establish wheather hCMEC/D3 cells are a suitable in vitro human BBB model of neuroinflammation.
Project description:Endothelial cells (ECs) are constantly submitted in vivo to hemodynamical forces derived from the blood circulation, including shear stress (SS). EC are able to detect SS and consequently adapt their phenotype, thus affecting many endothelial functions. If a plethora of shear stress-regulated molecular networks have been described in peripheral EC, less is known about the molecular responses of microvascular brain ECs which constitute the blood-brain barrier (BBB). In this work, we investigated the response of human cerebral microvascular ECs to laminar physiological shear stress using the well characterized hCMEC/D3 cell line. Interestingly, we showed that hCMEC/D3 cells responded to shear stress by aligning perpendicularly to the flow direction, contrary to peripheral endothelial cells which aligned in the flow direction. Whole proteomic profiles were compared between hCMEC/D3 cells cultured either in static condition or under 5 or 10 dyn.cm-2 SS for three days. 3592 proteins were identified and expression levels were significantly affected for 3% of them upon both SS conditions. Pathway analyses were performed which revealed that most proteins overexpressed by SS refer to the antioxidant defense, probably mediated by activation of the NRF2 transcriptional factor. Regarding down-regulated proteins, most of them participate to the pro-inflammatory response, cell motility and proliferation. These findings confirm the induction of EC quiescence by laminar physiological SS and reveal a strong neuroprotective effect of SS on hCMEC/D3 cells, suggesting a similar effect on the BBB. Our results also showed that SS did not significantly increase expression levels nor did it affect the localization of junctional proteins or the functional activity of several ABC transporters (P-glycoprotein and MRPs). This work provides new insights on the response of microvascular brain EC to SS and on the importance of SS for optimizing in vitro BBB models.
Project description:Transcriptional profiling of hCMEC/D3 endothelial cells after six hour incubation with conditioned media from unstimulated or LPS-stimulated monocyte derived macrophages
Project description:Increase in frequency and size of wildfire raises a significant public health concern due to rapid climate change. Epidemiological studies have shown that wildfire smoke exposure can increase the risk of neurological disorders. Our study aims to understand how wildfire smoke particles affect brain endothelial cells using both primary human brain microvascular endothelial cells (HBMEC) and an immortalized human cerebral microvascular endothelial cell line (hCMEC/D3). HBMEC and hCMEC/D3 were exposed to different levels of smoldering eucalyptus smoke particles (10, 30, 50 ug/mL) for 24 hr. Supernatants were collected for LDH and ELISA array, cell lysates were collected for RNAseq, and cells were fixed and stained for tight junction marker (Zonula Occludens-1, ZO-1) via ICC. Our study found that treatment to smoldering eucalyptus particles can increase inflammatory cytokine production dose-dependently in HBMECs and hCMEC/D3. Moreover, RNAseq analyses resulted in dose-dependent changes in HBMEC and hCMEC/D3 from wood smoke treatment that could be mediated via AhR and Nrf2 pathways.
Project description:The experiment was designed to look at the effect (on gene expression) of exposing human cerebromicrovascular endothelial cell line hCMEC/D3 to propionate (3 micromolar) for 24 h, to determine if circulating metabolites have the potential to affect integrity of the blood-brain barrier.
Project description:hCMEC/D3 cell line can be cultured in 2 distinct commercially available media. The objective here was to determine the differences such media induced.
Project description:Stroke is a leading cause of long-term disability worldwide. Patients with good recovery after ischemic stroke present a higher downregulation of PATJ comparing with patients with marked disability. To understand the effects of PATJ downregulation we generated PATJ-knockdown human microvascular ECs. We used microarrays to detail the gene expression to identify the modulated genes due to the depletion of the PATJ gene.
Project description:Some studies have revealed that treating chronic subdural hematoma (CSDH) with drugs is effective; however, the specific mechanism of this process has been seldom discussed. Herein, we sought to identify the potential underlying mechanism of atorvastatin and low-dose dexamethasone for CSDH treatment. We sequenced the hCMEC/D3 cell lines treated with single statin, dexamethasone and low dose dexamethasone after hematoma injuried and statin to find out the key point.