摘要
目的:航天飞行性贫血现象的机制尚不清楚。本实验利用第4代细胞旋转培养系统(RCCS-4)模拟微重力,研究其对促红细胞生成素(EPO)诱导培养K562细胞向红系分化的影响及其机制。方法:实验分为地面培养组(Control组)、EPO诱导组(EPO组)、模拟微重力组(MG组)和模拟微重力+EPO组(MG+EPO组),利用RCCS-4模拟微重力,分别在地面和模拟微重力环境下,采用EPO诱导培养K562细胞。采用联苯胺染色法观察各组K562细胞向红系分化情况;流式细胞术检测K562细胞CD71抗原表达水平,Western-blotting方法检测K562细胞丝氨酸/苏氨酸蛋白(AKT)及红系特异核蛋白转录因子-1(GATA-1)表达水平,流式细胞术检测K562细胞凋亡情况。结果:联苯胺染色阳性率比较,MG组和MG+EPO组明显低于EPO组(P<0.01);CD71抗原表达水平为MG+EPO组明显低于EPO组(P<0.01);AKT/GATA-1蛋白表达量MG组低于Control组,MG+EPO组低于EPO组(P<0.01);各组K562细胞凋亡率检测发现MG组和MG+EPO组(70.25±0.17%)较Control组和EPO组明显增加(P<0.01)。结论:模拟微重力可以通过下调红系分化相关蛋白AKT及GATA-1表达和诱导细胞凋亡来抑制EPO诱导K562细胞向红系分化。
Objective：The reason of spaceflight anemia is still unknown.We use the rotary cell culture system（RCCS-4） to invest the effect and mechanism of simulated microgravity on the hematopoietic differentiation of K562 cells induced by EPO.Method：RCCS-4 used to simulate microgravity,K562 cells were cultured in the ground（control）,ground with EPO（EPO）,simulated microgravity（MG）,simulated microgravity with EPO（MG＋EPO） respectively.Cell erythroid differentiation was observed by using benzidine staining method,CD71 expression was examined by using flow cytometric technology,AKT and GATA-1 expression intracellular were detected with western-blotting,cell apoptosis was also compared between the ground group and the simulated microgravity group.Results：Benzidine-positive rate in control,EPO,MG,MG＋EPO groups were 5.17±0.31%,24.41±0.59%,6.17±0.31%,4.00±0.37% respectively（P0.01）,after cultured 12h,CD71 expression level in EPO and MG＋EPO groups were 62.65±1.34% and 37.84±1.45%（P0.01）;after cultured 24h,the expression of CD71 in EPO group and MG＋EPO group were 69.24±2.06% and 55.78±1.70%（P0.01）.AKT and GATA-1 were significantly decreased in simulated microgravity group compared with the ground group.There were also found that cell apoptosis rate in control,EPO,MG,MG＋EPO groups were 34.17±1.06%,25.71±1.59%,65.91±0.84%,70.25±0.17%（P0.01）.Conclusion：Simulated microgravity can down regulate the AKT and GATA-1 expression and induce cell apoptosis to inhibit erythroid differentiation of K562 induced by EPO.
出处
《微循环学杂志》
2012年第2期24-27,F0003,I0001,共6页
Chinese Journal of Microcirculation
基金
国防科工委专项课题分题(B3320062101)
