| Title | 7-Methylxanthine Influences the Behavior of ADORA2A-DRD2 Heterodimers in Human Retinal Pigment Epithelial Cells |
| Authors | Fan, Yuzhuo Li, Jiarui Huang, Lvzhen Wang, Kai Zhao, Mingwei |
| Affiliation | Peking Univ, Hlth Sci Ctr, Inst Med Technol, Beijing, Peoples R China Peking Univ Peoples Hosp, Dept Ophthalmol, Beijing, Peoples R China Peking Univ Peoples Hosp, Clin Ctr Optometry, Beijing, Peoples R China Peking Univ, Hlth Sci Ctr, Coll Optometry, Beijing, Peoples R China Peking Univ Peoples Hosp, Eye Dis & Optometry Inst, Beijing, Peoples R China Beijing Key Lab Diag & Therapy Retinal & Choroid D, Beijing, Peoples R China |
| Keywords | FORM DEPRIVATION MYOPIA DOPAMINE RECEPTORS KINASE SCLERA |
| Issue Date | Dec-2022 |
| Publisher | OPHTHALMIC RESEARCH |
| Abstract | Introduction: The goal of this study was to investigate the presence of ADORA2A-DRD2 heterodimers in human retinal pigment epithelial (RPE) cells; determine if 7-methylxanthine (7-MX), a nonselective adenosine receptor antagonist which was used to control myopia progression, can influence the behavior of RPE cells through the ADORA2A-DRD2 receptor pathway; and assess the changes in the expression of signaling molecules during cellular signal transduction. Methods: Human RPE cells were cultured in vitro in the presence or absence of 7-MX. Cell proliferation was evaluated with the CCK-8 assay. Apoptosis and necrosis rates were determined by annexin V-FITC/propidium iodide staining and flow cytometry. Immunofluorescence and coimmunoprecipitation were used to examine the protein expression and colocalization of ADORA2A and DRD2 in RPE cells. ADORA2A and DRD2 were knocked down with small interfering RNAs (siRNAs). Changes in the protein expression of ERK1/2 and phospho-ERK1/2 (pERK 1/2), which are signaling molecules downstream of dopamine receptors, were evaluated by Western blot analysis. Results: Immunofluorescence and coimmunoprecipitation showed that ADORA2A and DRD2 were colocalized in RPE cells. The expression of ADORA2A in RPE cells was inhibited by treatment with 50 mu mol/L 7-MX for 48 h, and the expression of DRD2, ERK1/2, and pERK1/2 was increased after treatment with 50 mu mol/L 7-MX for 48 h. After siRNA-mediated knockdown of DRD2 in RPE cells and further treatment with 50 mu mol/L 7-MX for 48 h, the expression of DRD2 was nearly restored to the level observed in the native control. At the experimental concentrations, 7-MX and siRNAs did not affect the proliferation or apoptosis of human RPE cells. Conclusions: ADORA2A and DRD2 heterodimers were present in RPE cells. 7-MX may affect the behaviors of RPE cells through the ADORA2A-DRD2 receptor pathway. 7-MX is an inhibitor of ADORA2A receptors that can prevent inhibition of the DRD2 receptor pathway and increase DRD2 receptor pathway activity. This phenomenon may explain the mechanism of action through which 7-MX can control myopia progression. |
| URI | http://hdl.handle.net/20.500.11897/661857 |
| ISSN | 0030-3747 |
| DOI | 10.1159/000525563 |
| Indexed | SCI(E) |
| Appears in Collections: | 人民医院 |
