The regulating affects of receptor interacting protein 3 on retina ganglion cell-5 necroptosis following elevated hydrostatic pressure

International Conference and Exhibition on Molecular Medicine and Diagnostics

August 24-26, 2015 London, UK

Lei Shang, Dan Chen, Le Ping Zeng, Tu Hu, Lan Li, Jia Luo, Kun Xiong and JuFang Huang

Posters-Accepted Abstracts: J Mol Genet Med

Abstract :

Necroptosis is an important neuronal death mode in retinal ischemia, but the mechanism still needs clarify. RIP3 is characterized
as an N-terminal Serine/Threonine kinase, which participates in cell death signaling. Previous studies indicated RIP3 may
participate in neuronal necroptosis, and the activation of caspase-8 could cleave RIP3 to inactive form. In the present study, we
explored the effects of RIP3 in retinal necroptosis following elevated hydrostatic pressure (EHP) and discussed the possible role of
caspase-8 on regulation of RIP3 activity. Necrosis levels detection were repeated with pretreatment of Nec-1of 24 h to confirm the
existence of necroptosis. The expression of RIP3, downstream molecules in the pathway of RIP3-induced necroptosis and necrosis
levels of RGC-5 cells were detected by immunoblotting, immunofluorescence and flow cytometry at 6 h, 12 h or 24 h after EHP. Then,
RNAi to rip3 was used for further confirming RIP3’s effects on retinal necroptosis. Finally, caspase-8 inhibitor and activity peptide
were applied to try to unveil the regulated mechanism of RIP3 activity. The results showed that, RIP3 expression was up-regulated and
RIP3 enhance-labeled cells were coexisted with PI-positive cells after injury. PI-positive cells were reduced and ratios of necrosis were
decreased after injury when treating with Nec-1 and rip3 RNAi. The ROS and PYGL levels in pathway of RIP3-induced necroptosis
had been found to be decreased after rip3 knockdown. Caspase-8 inhibitor and activity peptide usage affected ratio of necrosis and
levels of ROS or PYGL. Our results indicated RIP3 participated in RGC-5 necroptosis following EHP and caspase-8 may interference
RIP3-induced necroptosis.

Biography :

Lei Shang has completed his PhD from Central South University. He is the research worker of Hunan Cancer Hospital in the department of Translational Medicine Research,
a premier Cancer Research Institute in Central South Region of China. He has published more than 10 papers in reputed journals.