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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 the effects of cinnamaldehyde on the leukemia K562 cell line by in vitro absorption model of Caco-2 cells and to differentiate K562 cells into myeloid and erythroid lineage. Methods Cinnamic aldehyde was transplanted into Caco-2 cell by Transwell transporter. The cytotoxic concentration was determined as 0, 50, 100, 200, 400, 600, 800 and 1000 μg · mL -1. The components of the model were determined by HPLC. The cytotoxic concentration of cinnamic aldehyde on K562 cells for 72 h was determined by MTT assay. C57BL / 6 C57BL / 6 mice were cultured with K562 (50,75μg · mL -1) for 72 h. The proportion of surface antigens CD235a, CD36, CD41, CD61, CD13, CD33 and CD14 in K562 cells was determined by flow cytometry . Results Cinnamaldehyde non-cytotoxic concentration of 200μg · mL -1, the model through the composition of cinnamaldehyde. The inhibitory rates of K562 cells treated with 50 and 75 μg · mL -1 cinnamaldehyde were (25.29 ± 0.97)% and (36.60 ± 0.18)% at 24 h and (48.23 ± 0.63)% at 48 h, (57.15 ± 0.58)% and 72 h (58.23 ± 0.63)%, (57.15 ± 0.58)%, respectively. Compared with the control group, the inhibitory effect was significant (P <0.05). The expression of myeloid differentiation phenotype CD13, CD33 and CD36 on K562 cells were (0.33 ± 0.21)% and (32.89 ± 0.19)%, respectively at 50 and 75μg · mL -1 cinnamic aldehyde for 72 h. , (7.73 ± 0.57)%, (0.72 ± 0.43)%, (38.80 ± 0.03)% and (10.90 ± 0.82)%, respectively, which were significantly higher than those in the control group (P <0.05). The expression of CD235a in erythroid differentiation phenotype was (52.38 ± 0.65)%, (57.48 ± 0.70)%, which was significantly higher than that in control group (P <0.05). There was no significant difference in the expression rates of megakaryocytic differentiation phenotypes CD41 and CD61 (P> 0.05). Conclusion Cinnamaldehyde can penetrate the in vitro absorption model of Caco-2 cells and differentiate K562 cells into myeloid and erythroid cells.