摘要
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 saponins of panax ginseng （TSPG） on the expression of erythropoietin receptor （EPOR） in K562 cells. Methods： The effect of TSPG on EPOR protein positive ratio in K562 cells was detected by using flow cytometry; the expression of EpoR protein in K562 cells was detected by immunocytochemistry; and EPOR protein in whole cell extracts, cytosol fractions and extractions of total cellular membrane protein of K562 cells were detected by Western Blot analysis. Results： EpoR protein positive ratio in K562 cell membrane was markedly decreased with the increase of the concentration of TSPG. K562 cell membrane in the control group was darkly stained, cytoplast weakly and the ratio of nucleus/cytoplasm was high; after being stimulated by 200 μg/ml TSPG for 72 h, K562 cell membrane was weakly stained, and cytoplast was abundant and darkly stained; and the ratio of nucleus/cytoplasm was markedly decreased. The expression of EPOR protein in whole cell extracts of K562 cells had no significant change with the increase of TSPG concentration. Compared with the control group, the expression of EpoR protein in total cellular membrane protein extracted from K562 cells induced by 200 μg/ml TSPG was weakened. The expression of EPOR protein in cytosol fraction of K562 cells induced by TSPG（100, 200, 300 and 400 mg/L） for 72 h increased with the increase of TSPG concentration. Conclusion： TSPG can change the location of EPOR in K562 cell, which provides new evidence for mechanism by which TSPG induces K562 cells to differentiate into erythroid lineage cells.
出处
《解剖学杂志》
CAS
CSCD
北大核心
2008年第1期22-24,共3页
Chinese Journal of Anatomy
基金
国家自然科学基金(30472253)
重庆市教委科学基金(KJ050303)
重庆医科大学创新基金
重庆医科大学博士启动基金
