scholarly journals High Glucose Induced Changes in Human VEC Phenotype in a 3D Hydrogel Derived From Cell-Free Native Aortic Root

2021 ◽  
Vol 8 ◽  
Author(s):  
Sergiu Cecoltan ◽  
Letitia Ciortan ◽  
Razvan D. Macarie ◽  
Mihaela Vadana ◽  
Andreea C. Mihaila ◽  
...  
Keyword(s):  
Aortic Valve ◽  
Aortic Root ◽  
High Glucose ◽  
Intercellular Junctions ◽  
Endothelin Receptors ◽  
Early Diabetes ◽  
Mesenchymal Transition ◽  
Inflammatory Phenotype ◽  
Endothelial To Mesenchymal Transition ◽  
Induced Changes

Background: Valvular endothelial cells (VEC) have key roles in maintaining valvular integrity and homeostasis, and dysfunctional VEC are the initiators and major contributors to aortic valve disease in diabetes. Previous studies have shown that HG stimulated an inflammatory phenotype in VEC. Inflammation was shown to induce endothelial to mesenchymal transition (EndMT), a process extensively involved in many pathologies, including calcification of the aortic valve. However, the effect of HG on EndMT in VEC is not known. In addition, there is evidence that endothelin (ET) is a proinflammatory agent in early diabetes and was detected in aortic stenosis, but it is not known whether HG induces ET and endothelin receptors and whether endothelin modulates HG-dependent inflammation in VEC. This study aims to evaluate HG effects on EndMT, on endothelin and endothelin receptors induction in VEC and their role in HG induced VEC inflammation.Methods and Results: We developed a new 3D model of the aortic valve consisting of a hydrogel derived from a decellularized extracellular cell matrix obtained from porcine aortic root and human valvular cells. VEC were cultured on the hydrogel surface and VIC within the hydrogel, and the resulted 3D construct was exposed to high glucose (HG) conditions. VEC from the 3D construct exposed to HG exhibited: attenuated intercellular junctions and an abundance of intermediate filaments (ultrastructural analysis), decreased expression of endothelial markers CD31 and VE–cadherin and increased expression of the mesenchymal markers α-SMA and vimentin (qPCR and immunocytochemistry), increased expression of inflammatory molecules ET-1 and its receptors ET-A and ET-B, ICAM-1, VCAM-1 (qPCR and Immunocytochemistry) and augmented adhesiveness. Blockade of ET-1 receptors, ET-A and ET-B reduced secretion of inflammatory biomarkers IL-1β and MCP-1 (ELISA assay).Conclusions: This study demonstrates that HG induces EndMT in VEC and indicates endothelin as a possible target to reduce HG-induced inflammation in VEC.

scholarly journals Cell-Type Transcriptome Atlas of Human Aortic Valves Reveal Cell Heterogeneity and Endothelial to Mesenchymal Transition Involved in Calcific Aortic Valve Disease

2020 ◽  
Vol 40 (12) ◽  
pp. 2910-2921
Author(s):  
Kang Xu ◽  
Shangbo Xie ◽  
Yuming Huang ◽  
Tingwen Zhou ◽  
Ming Liu ◽  
...  
Keyword(s):  
Aortic Valve ◽  
Single Cell ◽  
Aortic Valve Disease ◽  
Valve Disease ◽  
Cell Heterogeneity ◽  
Calcific Aortic Valve Disease ◽  
Mesenchymal Transition ◽  
Endothelial To Mesenchymal Transition ◽  
Human Aortic Valve ◽  
Insight Into

Objective: Leaflet thickening, fibrosis, and hardening are early pathological features of calcific aortic valve disease (CAVD). An inadequate understanding of the resident aortic valve cells involved in the pathological process may compromise the development of therapeutic strategies. We aim to construct a pattern of the human aortic valve cell atlas in healthy and CAVD clinical specimens, providing insight into the cellular origins of CAVD and the complex cytopathological differentiation process. Approach and Results: We used unbiased single-cell RNA sequencing for the high-throughput evaluation of cell heterogeneity in 34 632 cells isolated from 6 different human aortic valve leaflets. Cellular experiments, in situ localization, and bulk sequencing were performed to verify the differences between normal, healthy valves and those with CAVD. By comparing healthy and CAVD specimens, we identified 14 cell subtypes, including 3 heterogeneous subpopulations of resident valve interstitial cells, 3 types of immune-derived cells, 2 types of valve endothelial cells, and 6 novel valve-derived stromal cells found particularly in CAVD leaflets. Combining additional verification experiments with single-cell transcriptome profiling provided evidence of endothelial to mesenchymal transition involved in lesion thickening of the aortic valve leaflet. Conclusions: Our findings deconstructed the aortic valve cell atlas and suggested novel functional interactions among resident cell subpopulations. Our findings may provide insight into future targeted therapies to prevent CAVD.

High glucose induced endothelial to mesenchymal transition in human umbilical vein endothelial cell

2017 ◽  
Vol 102 (3) ◽  
pp. 377-383 ◽  
Author(s):  
Chun-Hong Yu ◽  
Suriguga ◽  
Meng Gong ◽  
Wen-Juan Liu ◽  
Ning-Xuan Cui ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
High Glucose ◽  
Umbilical Vein ◽  
Mesenchymal Transition ◽  
Human Umbilical Vein ◽  
Endothelial To Mesenchymal Transition

scholarly journals Resveratrol-Elicited PKC Inhibition Counteracts NOX-Mediated Endothelial to Mesenchymal Transition in Human Retinal Endothelial Cells Exposed to High Glucose

Antioxidants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 224 ◽  
Author(s):  
Roberta Giordo ◽  
Gheyath K. Nasrallah ◽  
Anna Maria Posadino ◽  
Francesco Galimi ◽  
Giampiero Capobianco ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
High Glucose ◽  
Ros Generation ◽  
Retinal Endothelial Cells ◽  
Mesenchymal Transition ◽  
Endothelial Markers ◽  
Fibrotic Process ◽  
Intracellular Ros ◽  
Endothelial To Mesenchymal Transition ◽  
Pre Treatment

Diabetes-associated long-term hyperglycaemia leads to oxidative stress-mediated fibrosis in different tissues and organs. Endothelial-to-mesenchymal-transition (EndMT) appears to play a role in diabetes-associated fibrotic conditions. Here, we investigate whether EndMT is implicated in the diabetic retinopathy fibrotic process and evaluate the possibility that resveratrol could counteract EndMT by inhibiting high glucose (HG)-induced increases in ROS. Primary Human Retinal Endothelial Cells (HRECs) were either pre-treated for 24 h with 1 µM resveratrol or left untreated, then glucose (30 mM) was applied at 3-day intervals for 10 days. qRT-PCR and ELISA were used to detect mRNA or protein expression of endothelial markers (CD31, CDH5, vWF) or mesenchymal markers (VIM, αSMA and collagen I), respectively. Intracellular ROS levels were measured with carboxy-DCFDA, while NOX-associated ROS levels were evaluated using the NADPH-specific redox biosensor p47-roGFP. Treatment of HRECs with HG increased intracellular ROS levels and promoted phenotype shifting towards EndMT, evidenced by decreased expression of endothelial markers concomitant with increased expression of mesenchymal ones. HG-induced EndMT appears to be mediated by NADPH-associated ROS generation as pre-treatment of HRECs with resveratrol or the NADPH inhibitor, diphenyleneiodonium chloride (DPI), attenuated ROS production and EndMT transition, suggesting that the effect of resveratrol on HG-induced ROS occurs via down-regulation of NADPH oxidase. It is worth noting that resveratrol or Chelerythrine, a Protein kinase C (PKC) inhibitor, reduce ROS and EndMT in HG-exposed cells, suggesting that NADPH activation occurs via a PKC-dependent mechanism. Taken together, our results provide the basis for a resveratrol-based potential protective therapy to prevent diabetic-associated complications.

Sign in / Sign up

Export Citation Format

Share Document