| Title |
Assessment of HIF-1α expression and release following endothelial injury in-vitro and in-vivo
|
|---|---|
| Published in |
Molecular Medicine, May 2018
|
| DOI | 10.1186/s10020-018-0026-5 |
| Pubmed ID | |
| Authors |
Lamia Heikal, Pietro Ghezzi, Manuela Mengozzi, Gordon Ferns |
| Abstract |
Endothelial injury is an early and enduring feature of cardiovascular disease. Inflammation and hypoxia may be responsible for this, and are often associated with the up-regulation of several transcriptional factors that include Hypoxia Inducible Factor-1 (HIF-1). Although it has been reported that HIF-1α is detectable in plasma, it is known to be unstable. Our aim was to optimize an assay for HIF-1α to be applied to in vitro and in vivo applications, and to use this assay to assess the release kinetics of HIF-1α following endothelial injury. An ELISA for the measurement of HIF-1α in cell-culture medium and plasma was optimized, and the assay was used to determine the best conditions for sample collection and storage. The results of the ELISA were validated using Western blotting and immunohistochemistry (IHC). In vitro, a standardized injury was produced in a monolayer of rat aortic endothelial cells (RAECs) and intracellular HIF-1α was measured at intervals over 24 h. In vivo, a rat angioplasty model was used. The right carotid artery was injured using a 2F Fogarty balloon catheter. HIF-1α was measured in the plasma and in the arterial tissue (0, 1, 2, 3 and 5 days post injury). The HIF-1α ELISA had a limit of detection of 2.7 pg/mL and was linear up to 1000 pg/ mL. Between and within-assay, the coefficient of variation values were less than 15%. HIF-1α was unstable in cell lysates and plasma, and it was necessary to add a protease inhibitor immediately after collection, and to store samples at -80 °C prior to analysis. The dynamics of HIF-1α release were different for the in vitro and in vivo models. In vitro, HIF-1α reached maximum concentrations approximately 2 h post injury, whereas peak values in plasma and tissues occurred approximately 2 days post injury, in the balloon injury model. HIF-1α can be measured in plasma, but this requires careful sample collection and storage. The carotid artery balloon injury model is associated with the transient release of HIF-1α into the circulation that probably reflects the hypoxia induced in the artery wall. |
Mendeley readers
The data shown below were compiled from readership statistics for 54 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 54 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 8 | 15% |
| Student > Bachelor | 7 | 13% |
| Student > Master | 7 | 13% |
| Researcher | 6 | 11% |
| Other | 6 | 11% |
| Other | 3 | 6% |
| Unknown | 17 | 31% |
| Readers by discipline | Count | As % |
|---|---|---|
| Medicine and Dentistry | 12 | 22% |
| Biochemistry, Genetics and Molecular Biology | 10 | 19% |
| Agricultural and Biological Sciences | 2 | 4% |
| Computer Science | 1 | 2% |
| Nursing and Health Professions | 1 | 2% |
| Other | 8 | 15% |
| Unknown | 20 | 37% |
Attention Score in Context
This research output has an Altmetric Attention Score of 3. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 17 September 2020.
All research outputs
#7,564,023
of 23,072,295 outputs
Outputs from Molecular Medicine
#369
of 1,152 outputs
Outputs of similar age
#130,813
of 327,769 outputs
Outputs of similar age from Molecular Medicine
#7
of 15 outputs
Altmetric has tracked 23,072,295 research outputs across all sources so far. This one is in the 44th percentile – i.e., 44% of other outputs scored the same or lower than it.
So far Altmetric has tracked 1,152 research outputs from this source. They typically receive more attention than average, with a mean Attention Score of 8.3. This one is in the 38th percentile – i.e., 38% of its peers scored the same or lower than it.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 327,769 tracked outputs that were published within six weeks on either side of this one in any source. This one has gotten more attention than average, scoring higher than 53% of its contemporaries.
We're also able to compare this research output to 15 others from the same source and published within six weeks on either side of this one. This one has gotten more attention than average, scoring higher than 53% of its contemporaries.