GATA-1对高原红细胞增多症模型大鼠骨髓CD71^+细胞EpoR表达的影响  

Influence of GATA-1 on Expression of EpoR in Bone Marrow CD71^+ Cells of Rat Model with High Altitude Polycythemia

作　　者：刘芳[1] 丁瑾[1] 魏巍[2] 冯婷婷[2] 陈英 冀林华[2] 韩园芳 

机构地区：[1]青海大学医学院生物化学教研室,青海西宁810000 [2]青海大学附属医院血液科,青海西宁810000

出　　处：《中国实验血液学杂志》2016年第3期884-891,共8页Journal of Experimental Hematology

基　　金：国家自然科学基金资助项目(No.81441116);青海省科技厅(应用)基础研究计划项目(No.2012-Z-729);青海大学"123高层次人才培养工程"-中青年学术带头人基金;青海大学医学院中青年科研基金团队项目(2014-KT-1)

摘　　要：目的:探讨GATA-1对高原红细胞增多症(high altitude polycythemia,HAPC)模型大鼠骨髓CD71+细胞促红细胞生成素受体(EpoR)表达的影响。方法:选用雄性SD大鼠48只,随机分为对照和HAPC模型2组。在海拔4 300米自然环境下复制HAPC模型并采用骨髓细胞分类计数和血液学参数检测进行验证。应用密度梯度离心和免疫磁珠分选相结合的方法分选大鼠骨髓CD71+细胞后,应用流式细胞术和细胞免疫荧光观察其EpoR的细胞表面表达水平,Q-PCR和Western blot检测其GATA-1、EpoR mRNA和蛋白的表达变化。分别在常氧和低氧下培养CD71+细胞,筛选最佳干扰序列GATA-1 shRNA1后,转染96 h,Q-PCR和Western blot检测GATA-1、EpoR mRNA和蛋白的表达水平。结果:血液学参数和骨髓细胞分类计数结果显示,HAPC大鼠模型复制成功。与对照组比较,HAPC模型组骨髓CD71+细胞膜表面EpoR的表达增高(P<0.05),GATA-1、EpoR mRNA和蛋白的表达也明显增高(P<0.05)。相关性分析显示,两组GATA-1、EpoR mRNA和蛋白的表达均呈正相关(对照组,r=0.929,P<0.01,r=0.802,P<0.05;HAPC模型组,r=0.822,P<0.05,r=0.839,P<0.01)。GATA-1 shRNA1干扰两组骨髓CD71+细胞96 h后,EpoR mRNA和蛋白的表达低于阴性对照组(P<0.05),且在低氧条件下GATA-1和EpoR的表达仍高于相应的常氧状态。结论:GATA-1对EpoR表达的影响可能与HAPC的发生发展相关联。Objective: To explore the influence of GATA-1 on expression of EpoR in bone marrow CD71+cells of rat model w ith high altitude polycythemia( HAPC). Methods: Forty- eight male SD rats w ere randomly divided into normal control and HAPC model group. HAPC model w as established at the altitude of 4 300 meters in the natural environment,and verified by bone marrow cell counts and hematological parameters. M yeloid CD71+cells w ere separated by the density gradient centrifugation combined w ith magnetic activated cell sorting. The expression of EpoR on cell membrane w as detected by flow cytometry and cell immunofluorescence. The expression changes of GATA-1 and EpoR mRNA and protein w ere detected by Q-PCR and Western blot,respectively. CD71+cells w ere cultured under normoxia and hypoxia,respectively. After transfection for 96 h,the optimal interference sequence GATA-1 shRNA1 w as selected. And the mRNA and protein expression level of GATA-1 and EpoR w ere detected by Q-PCR and Western blot respectively. Results: The animal model w ith HAPC w as established successfully and comfirmed by the bone marrow cell counting and the hematologic parameters in comparison w ith that of the normal control. EpoR expression on the myeloid CD71+cell membrane in HAPC group w as significantly higher than that in normal control( P < 0. 05). The expression of GATA-1 and EpoR in myeloid CD71+cells of HAPC group w as higher than that in control group( P <0. 05). The mRNA and protein expression of GATA-1 and EpoR in tw o 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). How ever,the mRNA and protein expression of EpoR at normoxia and hypoxia w as significantly low er than that in negative control group after interfernce w ith GATA-1 shRNA1 for 96 h( P < 0. 05). And the expression of GATA-1 and EpoR under hypoxia w as higher than that in normoxia. Conclusion: The effect of GATA-1 on EpoR expression may be correlated w ith the pathogenesis of HAPC.

关 键 词：高原红细胞增多症 GATA-1 促红细胞生成素受体 SHRNA 

分 类 号：R[医药卫生] R594.3