论文部分内容阅读
目的探讨模拟微重力对CD34+细胞分化的影响及其调控机制。方法在旋转细胞培养系统(RCCS)模拟微重力和正常重力条件下培养CD34+细胞,流式细胞仪检测各组细胞的系特异性抗原比例,考察CD34+细胞分化的改变。同时,进行PI细胞凋亡染色检测,考察细胞凋亡情况。另外,采用实时荧光定量PCR检测短期培养中流式分选出的CD34+细胞中的转录因子GATA-1 mRNA的表达。结果 GlyA+在RCCS组中和1-g液态对照组的表达量分别是(22.21±3.02)%和(60.05±3.08)%;CD33+在RCCS组和1-g液态组中的表达量分别为(52.12±1.92)%和(18.87±1.41)%,差异均有统计学意义(P<0.05)。1-g液态对照组和1-g甲纤组的分化比例(GlyA+%=54.39%±2.86%,CD33+%=21.09%±3.19%)相似,差异无统计学意义。各组凋亡比例均在1%以下。实验组中的GATA-1 mRNA的相对表达量约是对照组表达量的20%。结论模拟微重力抑制CD34+细胞向各系血细胞的分化,尤其抑制其向红系细胞的分化。红系分化调控转录因子GATA-1 mRNA下降,推测其可能是模拟微重力导致红系分化减少的原因之一。
Objective To investigate the effects of simulated microgravity on the differentiation of CD34 + cells and its regulatory mechanism. Methods CD34 + cells were cultured in a rotating cell culture system (RCCS) under simulated microgravity and normal gravity conditions. Flow cytometry was used to detect the percentage of lineage-specific antigens in each group of cells, and the changes of CD34 + cell differentiation were investigated. At the same time, PI staining for apoptosis was examined to investigate the apoptosis. In addition, real-time fluorescent quantitative PCR was used to detect the expression of GATA-1 mRNA in flow-sorted CD34 + cells in short-term culture. Results The expression levels of GlyA + in RCCS group and 1-g liquid control group were (22.21 ± 3.02)% and (60.05 ± 3.08)%, respectively. The expression levels of CD33 + in RCCS group and 1-g liquid group were ± 1.92)% and (18.87 ± 1.41)%, respectively, with statistical significance (P <0.05). There was no significant difference between the 1-g liquid group and the 1-g methylcellulose group (GlyA +% = 54.39% ± 2.86%, CD33 +% = 21.09% ± 3.19%). The percentage of apoptosis in each group was below 1%. The relative expression of GATA-1 mRNA in the experimental group was about 20% of the control group. Conclusion Simulated microgravity inhibits the differentiation of CD34 + cells to all the blood cells, especially the erythroid cells. Erythroid differentiation regulating transcription factor GATA-1 mRNA decreased speculated that it may be one of the reasons that reduce the differentiation of red line induced by microgravity.