人参总皂苷对K562细胞红细胞生成素受体表达的影响

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目的:探讨人参总皂苷(TSPG)对人红白血病细胞株(K562细胞)红细胞生成素受体(EpoR)表达的影响。方法:采用流式细胞仪检测TSPG作用前后的K562细胞膜EpoR阳性率的变化;免疫细胞化学检测TSPG作用前后K562细胞EpoR表达的变化;以免疫印迹法检测TSPG处理前后K562细胞全细胞、细胞膜和细胞质中EpoR含量的变化。结果:K562细胞膜表达EpoR蛋白的阳性率随TSPG剂量的增加而显著下降;免疫细胞化学染色可见对照组K562细胞胞膜着色深,胞质浅淡,核/质比例大;经TSPG 200mg/L作用72h后,K562细胞胞膜着色明显变浅,胞质着色明显加深且丰富,核/质比例明显降低;不同剂量的TSPG作用K562细胞72h后,全细胞总蛋白中的EpoR的表达无明显差异,经200mg/L TSPG诱导后的K562细胞膜中EpoR蛋白表达下降,而TSPG(100、200、300、400mg/L)诱导K562细胞72h后,胞质EpoR蛋白表达增强,且随着剂量的加大更加明显。结论:TSPG能使K562细胞的EpoR位置重塑,为研究TSPG诱导K562细胞向红系细胞分化的作用机制提供了依据。 Objective: To investigate the effect of total ginsenoside (TSPG) on the expression of erythropoietin receptor (EpoR) in human erythroleukemia cell line (K562 cells). Methods: Flow cytometry was used to detect the changes of EpoR positive rate of K562 cells before and after treatment with TSPG. Immunocytochemistry was used to detect the changes of EpoR expression in K562 cells before and after treatment with TSPG. The whole cells, cell membrane and cytoplasm of K562 cells before and after treatment with TSPG were detected by Western blotting. Changes in EpoR content. RESULTS: The positive rate of EpoR protein expression in K562 cell membrane decreased significantly with the increase of TSPG dosage. Immunocytochemical staining showed that the control group K562 cell membrane was dark in color, pale in cytoplasm, and had a large nuclear/cytoplasmic ratio; it was treated with TSPG 200 mg/L. After 72 hours, the cell membrane of K562 cells became lighter, the cytoplasm coloration was deeper and richer, and the ratio of nuclear to cytoplasm was significantly decreased. After 72 hours of different doses of TSPG in K562 cells, there was no significant difference in the expression of EpoR in total cell proteins. The expression of EpoR protein in K562 cell membrane was decreased after 200mg/L TSPG induction, but the expression of EpoR protein in cytoplasm was enhanced after TSPG (100, 200, 300, 400mg/L) induced K562 cells for 72h, and it increased with increasing dose. obvious. CONCLUSION: TSPG can remodel EpoR in K562 cells and provide a basis for studying the mechanism of TSPG-induced differentiation of K562 cells into erythroid cells.
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