GET THE APP

Stimulation of islet revascularization modulating HIF-1α expression using siPHD3
..

Transplantation Technologies & Research

ISSN: 2161-0991

Open Access

Stimulation of islet revascularization modulating HIF-1α expression using siPHD3


4th Global Surgery and Transplantation Congress

October 03-04, 2016 Atlanta, USA

A Langlois, E Maillard-Pedracini, M Prinz, R Beaurep?¨re, W Bietiger, C Sookhareea, C Mura, N Jeandidier, M Pinget and S Sigrist

University of Strasbourg, France

Posters & Accepted Abstracts: J Transplant Technol Res

Abstract :

Deleterious events for islet engraftment are related to insufficient islets revascularisation inducing ?² cells death. HIF-1?± is involved in various survival molecular pathways like cell proliferation and angiogenesis whereas for a chronic stabilization, it induces apoptosis. Prolyl-hydroxylase 3 (PHD3) mediates the degradation of HIF1?± under normal oxygen conditions. It could be interesting to find the best level of HIF-1?± expression in islets inhibiting PHD3 to induce angiogenesis while limiting apoptosis. The aim of this work was to realize a proof of concept of a new strategy of angiogenesis stimulation inhibiting PHD3 expression using siRNA on ?²-cell lines. RINm5F ?²-cells were lipofected (Lipofectamine RNAiMax) or not with 50 ?¼M of siPHD3. Apoptosis was determined using Apo-ONE?® Homogeneous Caspase-3/7 Assay, PHD3 expression by western blotting and VEGF secretion using Elisa kit 48 and 72 hours after lipofection. Caspase 3/7 activity is reduced with siPHD3, 48h (Control: 450?±18 RFU; siPHD3: 155?±12 RFU, p<0.001) and 72h post lipofection (Control: 689?±10 RFU; siPHD3: 213?±21 RFU, p<0.001). A significant decrease of PHD3 protein expression was observed using siPHD3 validating the efficiency of our approach (Control 48h: 0.48?±0.15 and siPHD3: 0.14?±0.03 PHD3 protein expression/?²-actin; p<0.01). Finally, siPHD3 shown a significant pro-angiogenic effect increasing VEGF secretion 72h post lipofection (Control: 9837?±470 and siPHD3: 17306?±2118 pg/mg of protein; p<0.05). Inhibition of PHD3 using siRNA, decreases apoptosis and increases VEGF secretion in RINm5F cells. Thus, modulation of HIF-1?± expression using lipofection of siRNA could increase islet vascularization and could be a realistic approach to improve graft survival.

Biography :

Email: a.langlois@ceed-diabete.org

50+ Million Readerbase

Recommended Conferences

International Conference and Expo on Heart and Cardio Care

Amsterdam, Netherlands

Journal Highlights

Google Scholar citation report
Citations: 209

Transplantation Technologies & Research received 209 citations as per Google Scholar report

Transplantation Technologies & Research peer review process verified at publons

Indexed In

Recommended Conferences

International Conference and Expo on Heart and Cardio Care

Amsterdam, Netherlands

International Conference and Expo on Heart and Cardio Care

Amsterdam, Netherlands
 
arrow_upward arrow_upward