高原红细胞增多症模型大鼠骨髓CD71^+细胞GATA-1和GATA-2表达的相关性

Expression relevance of GATA-1 and GATA-2 in bone marrow CD71^+ cells of high-altitude polycythemia rat model

目的:探讨高原红细胞增多症(HAPC)模型大鼠骨髓CD71+细胞转录因子GATA-1和GATA-2表达的相关性。方法:雄性SD大鼠48只,随机分为对照组和HAPC模型组。在海拔4 300 m自然环境下复制HAPC模型并采用骨髓涂片、骨髓细胞分类计数和血液学参数检测进行验证。流式细胞术检测骨髓CD71^+细胞数量相对变化趋势;RT-q PCR和Western blot法检测GATA-1和GATA-2的mRNA和蛋白表达变化,低氧培养CD71^+细胞,筛选最佳干扰序列GATA-1 shRNA1后,转染96 h,RT-q PCR和Western blot法检测GATA-1和GATA-2的mRNA和蛋白表达水平。结果:骨髓细胞分类计数、涂片及血液学参数检测结果显示,HAPC大鼠模型复制成功。与对照组比较,HAPC模型组骨髓CD71^+细胞明显增多,骨髓CD71^+细胞GATA-1的mRNA和蛋白表达增高,GATA-2的mRNA和蛋白表达差异无统计学显著性。对照组GATA-1和GATA-2的mRNA和蛋白表达呈负相关,HAPC模型组GATA-1和GATA-2的mRNA和蛋白表达无显著相关性。GATA-1 shRNA1干扰96 h后,GATA-1的mRNA和蛋白表达低于对照组,但是GATA-2的mRNA和蛋白表达变化差异无统计学显著性。结论:HAPC大鼠骨髓CD71+细胞GATA-1和GATA-2表达异常,两者的负相关关系改变可能是导致HAPC发生的机制之一。

AIM：To investigate the expression relevance of transcription factors GATA-1 and GATA-2 in the bone marrow CD71 ＋cells of a high-altitude polycythemia （HAPC） rat model.METHODS：Male SD rats （n=48） were randomly divided into normal control group and HAPC model group .HAPC model was established at an altitude of 4 300 m in the natural environment and verified by the morphology and quantity of the bone marrow cells and hematologic parameters detection .A relative change in the trend of bone marrow CD 71 ＋cell numbers was detected by flow cytometry analysis .The expression of GATA-1 and GATA-2 at mRNA and protein levels in the CD 71 ^＋cells was examined by RT-qPCR and West-ern blot .CD71 ^＋cells were cultured under hypoxic condition and transfected with the optimal interference sequence of GA -TA-1shRNA1 for 96 h.The expression of GATA-1 and GATA-2 at mRNA and protein levels was detected by RT-qPCR and Western blot .RESULTS：The establishment of the animal model with HAPC was successful as the bone marrow smears and the hematologic parameters showed compared with the control .The quantity of the bone marrow CD 71 ^＋cells of HAPC rats were significantly increased and the expression of GATA-1 at mRNA and protein levels in the CD 71 ^＋cells were higher than those of the control .The expression of GATA-2 at mRNA and protein levels was similar to that of the control .The correlation analysis showed that the expression of GATA-1 was negatively correlated with that of GATA-2 in the control, while no obvious correlation between them was observed in the HAPC rats .The expression of GATA-1 at mRNA and protein levels in HAPC group was lower than that in control group after interfered by GATA-1 shRNA1 for 96 h, but no obvious diversity of GATA-2 expression between the 2 groups was observed .CONCLUSION：GATA-1 and GATA-2 are abnormally expressed and their negative correlation is destroyed in HAPC , which may be one of the pathogenesis of HAPC .