G protein-coupled receptor 17 is involved in CoCl-induced hypoxic injury in RGC-5 cells.
- Author:
Kana LIN
1
;
Meili LIN
2
;
Yingfen GU
1
;
Shunguo ZHANG
1
;
Shiying HUANG
1
Author Information
1. Department of Pharmacy, Shanghai Children's Medical Center Affiliated to School of Medicine, Shanghai Jiaotong University, Shanghai 200127, China.
2. Department of Pharmacy, the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310009, China.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
Cell Hypoxia;
genetics;
Cell Line;
Cell Survival;
Cobalt;
Gene Expression Regulation;
drug effects;
Gene Knockdown Techniques;
Humans;
Hypoxia;
chemically induced;
genetics;
Receptors, G-Protein-Coupled;
genetics;
metabolism;
Retinal Ganglion Cells;
drug effects
- From:
Journal of Zhejiang University. Medical sciences
2018;47(5):487-492
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effect of G protein-coupled receptor 17 (GPR17) on hypoxia injury in retinal ganglion cells .
METHODS:CoCl (400 μmol/L) was used to induce hypoxic injury in RGC-5 cells. The expression of GPR17 and the effect of GPR17 ligands were investigated, and the role of GPR17 in hypoxia injury was further studied by transfection of RGC-5 cells with GPR17 small interfering RNA (siRNA). The cell viability was determined by MTT and the cell apoptosis rate was detected by flow cytometry analysis. The expression of GPR17 mRNA was determined with RT-PCR.
RESULTS:mRNA expressions of GPR17 in RGC-5 cells with and without CoCl treatment were 0.36±0.05 and 0.26±0.08(<0.01). Compared with hypoxia without any treatment, pretreatment with GPR17 agonists (LTD, UDP, UDP-G) significantly reduced cell viability (the survival rates of cells decreased by 29.6%, 31.8% and 33.9%, all <0.01), while the effect of GPR17 antagonist (cangrelor) was the opposite (the survival rates of cells increased by 33.2%, <0.01). Transfection with GPR17 SiRNA inhibited hypoxia-induced up-expression of GPR17 mRNA (<0.01)and reduced cell apoptosis[rates of cell apoptosis were(39.73±2.06)%,(42.50±3.64)% and (24.98±2.16)% for blank control, NC siRNA and GPR17 siRNA groups, <0.01].
CONCLUSIONS:GPR17 may mediate hypoxia injury in RGC-5 cells, while the knockdown of GPR17 can reduce the hypoxia injury.
