桂皮醛通过Caco-2细胞体外吸收模型对白血病K562细胞株的作用

Effects of Cinnamic aldehyde on the leukemia cell line K562 using Caco-2 cells in vitro absorption model

目的研究桂皮醛通过Caco-2细胞体外吸收模型作用于白血病K562细胞株并使K562细胞向髓系、红系分化的情况。方法桂皮醛经Transwell转运池Caco-2细胞模型,确定无细胞毒质量浓度为0,50,100,200,400,600,800,1000μg·mL^(-1),高效液相色谱法检测透过模型的成分。噻唑蓝(MTT)法检测桂皮醛作用K562细胞72 h的无细胞毒的浓度范围。桂皮醛标准品(50,75μg·mL^(-1))直接培养白血病细胞K562共72 h,流式细胞仪检测K562细胞的表面抗原CD235a、CD36、CD41、CD61、CD13、CD33和CD14的比例。结果桂皮醛无细胞毒质量浓度为200μg·mL^(-1),透过模型的成分为桂皮醛。50,75μg·mL^(-1)桂皮醛作用K562细胞在以下不同时间的抑制率:24 h为(25.29±0.97)%,(36.60±0.18)%;48 h为(48.23±0.63)%,(57.15±0.58)%;72 h为(58.23±0.63)%,(57.15±0.58)%,和对照组相比,抑制效果明显(P<0.05)。50,75μg·mL^(-1)桂皮醛作用K562细胞72 h后,髓系分化表型CD13、CD33、CD36在K562细胞上表达量分别为(0.33±0.21)%,(32.89±0.19)%,(7.73±0.57)%;(0.72±0.43)%,(38.80±0.03)%,(10.90±0.82)%,和对照组比较,表达均明显增高(P<0.05)。红系分化表型CD235a的表达量为(52.38±0.65)%,(57.48±0.70)%,和对照组比较,表达均明显增高(P<0.05)。巨核系分化表型CD41、CD61的表达率无明显变化(P>0.05)。结论桂皮醛能透过Caco-2细胞体外吸收模型且能使K562细胞向髓系、红系分化。

Objective To study on the effects of Cinnamic aldehyde on leukemia cell line K562 by Caco-2 cells in vitro absorption model.Methods The effective components of cinnamon（ 0,50,100,200,400,600,800,1000 μg·mL^-1） were determined by Caco-2 cell model of Transwell,and the concentration was determined by HPLC. No cytotoxic concentration range of Cinnamic aldehyde acting on K562 cells for 72 h is detected by MTT assay. After 72 h incubation of Cinnamic aldehyde standard（ 50,75 μg·mL^-1） and leukemia K562 cells,the cells surface antigens including CD235 a,CD36,CD41,CD61,CD13,CD33 and CD14 were determined by Flow cytometry. Results The active ingredient of cinnamon is extracted by transwell transport pool of Caco-2 cell model and no cytotoxic concentration is 200 μg · mL^-1. The cinnamicaldehyde is the component which goes through the model by HPLC. The 24 h inhibition rates（ IRs） of Cinnamic aldehyde on K562 cells are（ 25. 29 ± 0. 97） % and（ 36. 60 ± 0. 18） % at the concentrations of 50 and 75 μg·mL^-1,respectively; IRs for 48 h are（ 48. 23 ± 0. 63） % and（ 57. 15 ± 0. 58） %; IRs for 72 h are（ 58. 23 ± 0. 63） % and（ 57. 15 ± 0. 58） %. Compared with the control group,the inhibitory activity is obvious（ P〈0. 05）. After incubation 72 h, the expressions of myeloid differentiation phenotypes including CD13, CD33, CD36 on K562 cells are（ 0. 33 ± 0. 21） %,（ 32. 89 ± 0. 19） %,（ 7. 73 ± 0. 57） % and（ 0. 72 ± 0. 43） %,（ 38. 80 ± 0. 03） %,（ 10. 90 ± 0. 82） % at the concentrations of 50 and 75 μg·mL^-1,respectively. Compared with the control group,the inhibition increased（ P〈0. 05）. The phenotypic expressions of erythroid differentiation are（ 52. 38 ± 0. 65） %,（ 57. 48 ± 0. 70） %. Compared with the control group,the inhibition increased（ P〈0. 05）. Megakaryocyte differentiated phenotype CD41,CD61 expression has no significant change（ P〉0. 05）. Conclusion The Cinnamic aldehyde can go through the Caco-2 in vitro absorption model and enables the K562 cells to differentiate into myeloid and erythroid.