title: [Influence of GATA-1 on Expression of EpoR in Bone Marrow CD71+ Cells of Rat Model with High Altitude Polycythemia].
abstract: OBJECTIVE
  To explore the influence of GATA-1 on expression of EpoR in bone marrow CD71+ cells of rat model with high altitude polycythemia (HAPC).
  
  
  METHODS
  Forty-eight male SD rats were randomly divided into normal control and HAPC model group. HAPC model was established at the altitude of 4 300 meters in the natural environment, and verified by bone marrow cell counts and hematological parameters. Myeloid CD71+ cells were separated by the density gradient centrifugation combined with magnetic activated cell sorting. The expression of EpoR on cell membrane was detected by flow cytometry and cell immunofluorescence. The expression changes of GATA-1 and EpoR mRNA and protein were detected by Q-PCR and Western blot, respectively. CD71+ cells were cultured under normoxia and hypoxia, respectively. After transfection for 96 h, the optimal interference sequence GATA-1 shRNA1 was selected. And the mRNA and protein expression level of GATA-1 and EpoR were detected by Q-PCR and Western blot respectively.
  
  
  RESULTS
  The animal model with HAPC was established successfully and comfirmed by the bone marrow cell counting and the hematologic parameters in comparison with that of the normal control. EpoR expression on the myeloid CD71+ cell membrane in HAPC group was significantly higher than that in normal control (P<0.05). The expression of GATA-1 and EpoR in myeloid CD71+ cells of HAPC group was higher than that in control group (P<0.05). The mRNA and protein expression of GATA-1 and EpoR in two groups positively correlated (control group, r=0.929, P<0.01, r=0.802, P<0.05; HAPC group, r=0.822, P<0.05, r=0.839, P<0.01). However, the mRNA and protein expression of EpoR at normoxia and hypoxia was significantly lower than that in negative control group after interfernce with GATA-1 shRNA1 for 96 h (P<0.05). And the expression of GATA-1 and EpoR under hypoxia was higher than that in normoxia.
  
  
  CONCLUSIONS
  The effect of GATA-1 on EpoR expression may be correlated with the pathogenesis of HAPC.
authors.affiliation: Department of Biochemistry, Medical College, Qinghai University, Xining 810000, Qinghai Province, China. E-mail: qhxnlf2006@163.com.
authors.forename: Fang
authors.initials: F
authors.lastname: Liu
authors.affiliation: Department of Biochemistry, Medical College, Qinghai University, Xining 810000, Qinghai Province, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Ding
authors.affiliation: Department of Hematology, Afiliated Hospital of Qinghai University, Xining 810000, Qinghai Province, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wei
authors.affiliation: Department of Hematology, Afiliated Hospital of Qinghai University, Xining 810000, Qinghai Province, China.
authors.forename: Ting-Ting
authors.initials: TT
authors.lastname: Feng
authors.affiliation: Department of Biochemistry, Medical College, Qinghai University, Xining 810000, Qinghai Province, China.
authors.forename: Yin
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Hematology, Afiliated Hospital of Qinghai University, Xining 810000, Qinghai Province, China.
authors.forename: Lin-Hua
authors.initials: LH
authors.lastname: Ji
authors.affiliation: Department of Hematology, Afiliated Hospital of Qinghai University, Xining 810000, Qinghai Province, China.
authors.forename: Yuan-Fang
authors.initials: YF
authors.lastname: Han
mesh_terms: D000531:Altitude; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D001854:Bone Marrow Cells / Q000378:metabolism*; D002469:Cell Separation; D004195:Disease Models, Animal; D005434:Flow Cytometry; D050982:GATA1 Transcription Factor / Q000378:metabolism*; D008297:Male; D011086:Polycythemia / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D017467:Receptors, Erythropoietin / Q000378:metabolism*; D011990:Receptors, Transferrin / Q000378:metabolism
chemical_list: D015703:Antigens, CD; C068126:CD71 antigen; D050982:GATA1 Transcription Factor; C494690:Gata1 protein, rat; D012333:RNA, Messenger; D017467:Receptors, Erythropoietin; D011990:Receptors, Transferrin