Influence of HIF-2α on the expression of GATA-1 in bone marrow CD71 + cell of high altitude polycythemia rat model
10.3760/cma.j.issn.0253-2727.2016.08.013
- VernacularTitle:HIF-2α对高原红细胞增多症模型大鼠骨髓CD71+细胞GATA-1表达的影响
- Author:
Fang LIU
1
;
Wei WEI
;
Jin DING
;
Ying CHEN
;
Tingting FENG
;
Linhua JI
;
Jiyu SHI
Author Information
1. 青海大学医学院生物化学教研室
- Keywords:
High altitude polycythemia;
Hypoxia-inducible factor-2 αlpha;
GATA-1;
Small interfering RNA
- From:
Chinese Journal of Hematology
2016;37(8):696-701
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the influence ofhypoxia-inducible factor-2 αlpha (HIF-2α) on the expression of erythroid-specific transcription factor GATA-1 in bone marrow CD71+ cells of rat model with high altitude polycythemia (HAPC).Methods A total of 48 male SD rats were selected and randomly divided into normal control group and HAPC group.HAPC model was established at an altitude of 4 300 meters in the natural environment and verified by bone marrow cell classification and counting,hematologic parameters and serum EPO detection.Bone marrow CD71 + cells were separated by a combination of methods with density gradient centrifugation and magnetic activated cell sorting.The changes of expression level of HIF-2α,GATA-1 mRNA and proteins were detected by Q-PCR and Western blot.CD71 + cells were cultured under hypoxia condition and transfected with selected optimal HIF-2α shRNAi3 for 96 h.And the expression level of HIF-2α and GATA-1 mRNA and proteins were detected by Q-PCR and Western blot.Results The results of bone marrow cell counts,the hematologic parameters and the serum EPO content showed that the HAPC rat model was successfully established.The expression of HIF-2α and GATA-1 mRNA and protein in bone marrow CD71 + cells of HAPC group was higher than that in control group (P＜0.05).And HIF-2α and GATA-1 of HAPC group were positively correlated at the expression levels of mRNA and protein,respectively (r=0.923,P＜0.01;r=0.838,P＜0.01).However,the expression of HIF-2α and GATA-1 mRNA and protein in HAPC group was significantly lower than that in control groups after interfered by HIF-2α shRNAi3 for 96 h (P＜0.05).Conclusion The effect of HIF-2α on GATA-1 expression may be correlated with the pathogenesis of HAPC.
