Effects of hypoxia-inducible factor -2α on the expression of tight junction proteins and permeability in rat glomerular endothelial cells under hypoxia condition
10.3760/cma.j.issn.1001-7097.2016.10.008
- VernacularTitle:低氧环境下低氧诱导因子2α对大鼠肾小球内皮细胞细胞间紧密连接蛋白表达和细胞通透性的影响
- Author:
Pengli LUO
;
Yanjun WANG
;
Yanyan YANG
;
Jiajia YANG
- Publication Type:Journal Article
- Keywords:
Cell hypoxia;
Basic Helix-Loop-Helix transcription factors;
Cell membrane permeability;
Tight junction proteins;
Hypoxia-inducible factor-2alpha
- From:
Chinese Journal of Nephrology
2016;32(10):766-771
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the role of hypoxia?inducible factor?2α(HIF?2α) in the expression of tight junction proteins and permeability alterations in rat glomerular endothelial cells (rGENCs) under hypoxia condition. Methods The expressions of the HIF?2α and tight junction proteins such as occludin and ZO?1 of rGENCs were examined after exposed to 5%oxygen at different treatment time periods (0 h, 12 h, 24 h and 48 h). Then lentiviral transfection was used to knock down HIF?2α expression in rGENCs. The cells were split into four groups, including i) control group where rGENCs were cultured under normal oxygen conditions, ii) hypoxia group, iii) negative control group where rGENCs were infected with a negative vector, iv) HIF?2α lentivirus transfection group. Group ii, iii and iv were kept in hypoxic chamber (5% O2, 5% CO2 and 90% N2) for 24 h. The expressions of occludin, ZO?1 and HIF?2α were assessed by Western blotting. The permeability of rGENCs was measured using trans?epithelium electrical resistant (TEER) by Millicell? ERS voltohmmeter. Results With the elongation of hypoxia time, the expression of HIF?2α was increased gradually, while the occludin expression was decreased, there was statistically significance difference in each group (all P<0.01). The expression of ZO?1 also decreased gradually under hypoxia circumstance, but no statistically significant was found between 24 h and 48 h groups (all P>0.05). And a dramatic decrease in TEER of hypoxia cells was detected as compare with control cells (P<0.01). After knockdown of HIF?2αexpression, both expressions of occludin and ZO?1 were increased significantly compared with hypoxia cells (P<0.01), and TEER elevated at the same time (P<0.01). Above indexes had no statistical difference between hypoxia cells and negative control cells (all P>0.05). Conclusion Hypoxia may promote HIF?2α expression, which could increase the permeability of rGENCs by reducing the expression of occludin and ZO?1.
