Effects of leptin on proliferation and differentiation of hypoxic rat retinal progenitor cells in vitro.
10.12122/j.issn.1673-4254.2022.03.06
- Author:
Yao XING
1
;
Zi Yao LIU
1
;
Xiao Hui ZHANG
1
;
Jian Ming WANG
1
Author Information
1. Department of Ophthalmology, Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710004, China.
- Publication Type:Journal Article
- Keywords:
cell differentiation;
cell proliferation;
hypoxia;
leptin;
retinal progenitor cells
- MeSH:
Animals;
Cell Differentiation/drug effects*;
Cell Hypoxia/drug effects*;
Cell Proliferation/drug effects*;
Cells, Cultured;
Leptin/pharmacology*;
PTEN Phosphohydrolase/metabolism*;
Rats;
Rats, Sprague-Dawley;
Retina/metabolism*;
Stem Cells/metabolism*;
Tubulin
- From:
Journal of Southern Medical University
2022;42(3):354-359
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the the effects of leptin on the proliferation, differentiation and PTEN expression of rat retinal progenitor cells (RPCs) cultured under hypoxic condition.
METHODS:SD rat RPCs were cultured in normoxic conditions or exposed to hypoxia in the presence of 0, 0.3, 1.0, 3.0, 10, and 30 nmol/L leptin for 12, 48 and 72 h, and the cell viability was assessed using cell counting kit 8 (CCK 8) assay. The RPCs in primary culture were divided into control group, hypoxia group, and hypoxia+leptin group, and after 48 h of culture, the cell medium was replaced with differentiation medium and the cells were further cultured for 6 days. Immunofluorescence staining was employed to detect the cells positive for β-tubulin III and GFAP, and Western blotting was used to examine the expression of PTEN at 48 h of cell culture.
RESULTS:The first generation of RPCs showed suspended growth in the medium with abundant and bright cellular plasma and formed mulberry like cell spheres after 2 days of culture. Treatment with low-dose leptin (below 3.0 nmol/L) for 48 h obviously improved the viability of RPCs cultured in hypoxia, while at high concentrations (above 10 nmol/L), leptin significantly suppressed the cell viability (P < 0.05). The cells treated with 3.0 nmol/L leptin for 48 h showed the highest viability (P < 0.05). After treatment with 3.0 nmol/L leptin for 48 h, the cells with hypoxic exposure showed similar GFAP and β-tubulin Ⅲ positivity with the control cells (P>0.05), but exhibited an obvious down-regulation of PTEN protein expression compared with the control cells (P < 0.05).
CONCLUSION:In rat RPCs with hypoxic exposure, treatment with low dose leptin can promote the cell proliferation and suppress cellular PTEN protein expression without causing significant effects on cell differentiation.
