Liu F, Wei W, Ding J, Chen Y, Feng T T, Ji L H, Shi J Y
Department of Biochemistry, Medical College, Qinghai University, Xi'ning 810000, China.
Zhonghua Xue Ye Xue Za Zhi. 2016 Aug 14;37(8):696-701. doi: 10.3760/cma.j.issn.0253-2727.2016.08.013.
To explore the influence of hypoxia-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).
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.
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).
The effect of HIF-2α on GATA-1 expression may be correlated with the pathogenesis of HAPC.
探讨缺氧诱导因子-2α（HIF-2α）对高原红细胞增多症（HAPC）大鼠模型骨髓CD71(+)细胞中红系特异性转录因子GATA-1表达的影响。
选取48只雄性SD大鼠，随机分为正常对照组和HAPC组。于海拔4300米自然环境中建立HAPC模型，并通过骨髓细胞分类计数、血液学参数及血清EPO检测进行验证。采用密度梯度离心和磁珠分选相结合的方法分离骨髓CD71(+)细胞。运用Q-PCR和Western blot检测HIF-2α、GATA-1 mRNA及蛋白表达水平的变化。将CD71(+)细胞置于缺氧条件下培养，转染筛选出的最佳HIF-2α shRNAi3 96小时。通过Q-PCR和Western blot检测HIF-2α和GATA-1 mRNA及蛋白表达水平。
骨髓细胞计数、血液学参数及血清EPO含量结果显示，HAPC大鼠模型成功建立。HAPC组骨髓CD71(+)细胞中HIF-2α和GATA-1 mRNA及蛋白表达高于对照组（P<0.05）。HAPC组中HIF-2α与GATA-1在mRNA和蛋白表达水平上分别呈正相关（r=0.923，P<0.01；r=0.838，P<0.01）。然而，经HIF-2α shRNAi3干扰96小时后，HAPC组中HIF-2α和GATA-1 mRNA及蛋白表达显著低于对照组（P<0.05）。
HIF-2α对GATA-1表达的影响可能与HAPC的发病机制相关。
