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A Rhodopsin Transport Assay by High-Content Imaging Analysis
Published on: January 16, 2019
PKM2 Is a Potential Diagnostic and Therapeutic Target for Retinitis Pigmentosa
Peiwen Zhu1, Qian Yang1, Gang Li1
1Department of Ophthalmology and Vision Science, Eye and Ear, Nose and Throat Hospital of Fudan University, Shanghai 200031, China.
Abstract:
Retinitis pigmentosa (RP) is a major cause of blindness that is difficult to diagnose and treat. PKM2, a subtype of pyruvate kinase, is strongly associated with oxidative stress and is expressed in photoreceptors. We investigated whether PKM2 reduces photoreceptor cell apoptosis and evaluated possible antiapoptotic mechanisms in RP. We established RP models by exposing 661W cells to blue light and modulated PKM2 activity using a PKM2 inhibitor. We measured the apoptosis rates using calcein-acetoxymethyl ester/propidium iodide double staining and Cell Counting Kit-8, the oxidative stress levels using a reactive oxygen species assay, and the changes in protein expression by western blotting. Photodamage increased PKM2 expression, cellular oxidative stress, and apoptosis of 661W cells. PKM2 inhibition significantly reduced the levels of apoptosis and oxidative stress induced by photodamage. Our data suggest that PKM2 is a potential disease marker and therapeutic target for RP.

