Activation of the sweet taste receptor T1R3 by sucralose attenuates VEGF-induced vasculogenesis in a cell model of the retinal microvascular endothelium

Abstract

BackgroundOne of the most prevalent microvascular complications for patients with diabetes is diabetic retinopathy (DR) associated with increased retinal endothelial blood vessel formation. Treatments to reduce vascularisation in the retinal endothelium are linked to improved sight in patients with DR. Recently, we have demonstrated the novel protective role of the artificial sweetener, sucralose, and the sweet taste receptor, T1R3, in the pulmonary endothelium to reduce vascular leak. In the present study, we examined the role of sucralose and sweet taste receptors on vasculogenic processes (proliferation, migration, adhesion and tube formation) in a cell model of the retinal endothelium.MethodsWe exposed human retinal microvascular endothelial cells (RMVEC) to VEGF as an in vitro model of DR in the presence and absence of T1R3 agonist sucralose.ResultsIn RMVEC, we observed increased VEGF-induced cell proliferation, migration, adhesion and tube formation, which was significantly attenuated by exposure to the artificial sweetener sucralose. Following siRNA knockdown of the sweet taste receptor, T1R3, but not T1R2, the protective effect of sucralose on VEGF-induced RMVEC vasculogenic processes was blocked. We further demonstrate that sucralose attenuates VEGF-induced Akt phosphorylation to protect the retinal microvasculature.ConclusionThese studies are the first to demonstrate a protective effect of an artificial sweetener, through the sweet taste receptor T1R3, on VEGF-induced vasculogenesis in a retinal microvascular endothelial cell line.Electronic supplementary materialThe online version of this article (10.1007/s00417-018-4157-8) contains supplementary material, which is available to authorized users.

Acute Exposure to Fructose Impairs Endothelium-Dependent Relaxation via Oxidative Stress in Isolated Rat Aortic Rings

“…Several studies indicate the possibly beneficial effect of some sweeteners for health [25, 26], but it is well known that fructose intake induces hypertension and impairs vascular contraction and relaxation [15, 27, 28]. In our study, high-fructose intake increased BP and body weight (Fig.…”

Lutein and zeaxanthin attenuates VEGF-induced neovascularisation in human retinal microvascular endothelial cells through a Nox4-dependent pathway

“…It is therefore vital to understand the molecular mechanisms which underlie neovascularisation in DR pathology, and use this information to develop therapeutic options for patients. We have previously identified the presence of novel GPCRs, T1R2/3, in human retinal microvascular endothelial cells, which are activated by a range of acutely sweet molecules (Lizunkova et al 2019). Our studies demonstrated that the sweetener, sucralose, blocked VEGF-induced angiogenic processes such as permeability and neovascularisation, in the human retinal microvasculature.…”

“…For the Nox4 activity kit (Cusabio, Texas, USA), the assay was performed as per the manufacturer's protocol and absorbance was measured at 450nm using a microplate reader (Tecan Sunrise). Angiogenic processes were measured as cell proliferation, migration, adhesion and neovascularisation as previously described (Lizunkova et al 2019). In brief, RMVEC were quiesced for 24 h with 1% FBS followed by exposure to bevacizumab, L, Z or combined treatment, in the presence and absence of VEGF, for 6 h. Alternatively, cells were exposed to bevacizumab, L, Z or combined treatment, with VEGF (50 ng/ml), in the presence of GLX7013114 (1 µM) (Raystar Bio, Hangzhou Guangyuan, China) or the vehicle control (DMSO).…”

Extracellular vesicles released from p18 overexpressing pulmonary endothelial cells are barrier protective – potential implications for acute respiratory distress syndrome

“…Cell viability of LMVEC transfected with miRNA mimics was assessed using CCK8 assay, as per the manufacturer’s guidelines, with absorbance read at 450 nm. 25…”

“…The pellet, comprising of EDEVs, was resuspended in sterile Dulbecco's PBS (DPBS) for permeability assays and quantification using qNano Gold (Izon Science) at 0.5 V, 45 mm stretch, 15 mbar pressure, using the NP200 pore and normalized to media as a control, RNAlater for miRNA analysis, or lysed for protein concentration analysis. 23,24 Permeability measurements Changes in monolayer resistance were measured using the electrical cell impedance sensor (ECIS) technique and fluorescein isothiocyanate (FITC) dextran permeability technique 25 as previously described. 17 Naı¨ve, un-transfected LMVECs were plated on collagen-coated ECIS arrays or Transwell inserts and cultured for 24 h, prior to exposure to freshly-prepared EDEVs, in the presence and absence of LPS (1 mg/ml).…”