Abstract
Purpose :
The aim of this study is to investigate the effects of Probucol on high glucose-induced expressions of specificity protein 1(SP1), kelchlike ECH associated protein1 (Keap1), NF-E2-related factor 2 (Nrf2) and glutamate-cysteine ligase catalytic (GCLC) in the cultured human Müller cells and explore the protective role of the Probucol on Müller cells in high glucose environment.
Methods :
Müller cells were cultured from human retinal tissue, and identified by their expression of glutamine synthetase(GS). The cells were randomly divided into four groups: normal glucose group (group A); normal glucose with Probucol (group B); high glucose group (group C); high glucose with Probucol (group D). The expression of SP1, Keap1, Nrf2, and GCLC was examined by immunocytochemitry. The messenger RNA (mRNA) expressions of SP1, Keap1, Nrf2, and GCLC were evaluated by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR).
Results :
More than 95% of the cultured cells expressed GS. Immunocytochemistry showed that SP1, Keap1, Nrf2, and GCLC proteins were all expressed by human Müller cells. qRT-PCR indicated that SP1, Keap1, and Nrf2 mRNAs in group C increased significantly compared with group A(P<0.05)and GCLC mRNA in group C decreased significantly compared with group A(P<0.050). However, SP1 and Keap1 mRNAs in group D decreased significantly compared with group C(P<0.05), Nrf2 and GCLC mRNAs in group D increased compared with group C(P<0.05).
Conclusions :
The expressions of SP1, Keap1, and Nrf2 were up-regulated, while the expression of GCLC was down-regulated by high glucose in cultured human Müller cells. Probucol can mitigate such effects of high glucose.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.