丹皮酚对人黑素瘤A375细胞增殖和凋亡的影响  

Effects of paeonol on the proliferation and apoptosis of A375 human melanoma cells

作　　者：陶玥[1] 张孟丽[2] 马鹏程[2] 孙建方[2] 周武庆[2] 包军[1] 

机构地区：[1]南京市鼓楼医院皮肤科,210008 [2]中国医学科学院北京协和医学院皮肤病研究所

出　　处：《中华皮肤科杂志》2014年第6期393-396,共4页Chinese Journal of Dermatology

摘　　要：目的探讨丹皮酚对体外培养人黑素瘤A375细胞增殖和凋亡的影响及其作用机制。方法CCK8法检测0．5，1，2，4，8mmol／L丹皮酚作用24，48和72h后A375细胞增殖水平。用0、1．25、2．5和5mmol／L浓度丹皮酚作用24h后，以Annexin-V／PI法观察A375细胞凋亡的变化、分别测定caspase3，caspase8和caspase9的活性，并以Western印迹检测p53，NF-κB及相关蛋白水平的变化。结果与空白对照组比较，0．5，1，2，4，8mmol／L丹皮酚作用24，48和72h后均对A375细胞的增殖有抑制作用，且与时间、浓度呈依赖性。1．25、2．5、5mmol／L丹皮酚在作用24h时，A375细胞早期凋亡率由对照组的（3．11±0．53）％分别上升至（13．74±1．73）％、（25．95±0．57）％、（46．44±0．81）％，各组与对照组间差异均有统计学意义（P〈0．05或〈0．01）。2．5mmol／L和5mmol／L作用组的caspase3，caspase8，caspase9活性升高，且与对照组间比较，差异均有统计学意义（P〈0．05或〈0．01）。p53及Bax的蛋白水平随药物浓度的增加而升高，NF-κB、bcl-2、bcl-XL的蛋白水平随药物浓度增加而降低。结论丹皮酚对黑素瘤A375细胞具有抑制增殖和诱导凋亡的作用。可通过细胞内和细胞外两条途径发挥促凋亡作用，其机制可能与调节p53及NF-κB基因有关。Objective To study the effect of paeonol on the proliferation and apoptosis of A375 human melanoma cells and its mechanism. Methods Cell counting kit-8 （CCK8） was used to evaluate the proliferative activity of A375 cells treated with paeonol of 0.5,1,2,4,8 mmol/L for 24, 48 and 72 hours respectively. Subsequently, A375 cells were treated with paeonol of 1.25, 2.5 and 5 mmol/L for 24 hours followed by double staining with annexin g and propidium iodide for the detection of cell apoptosis, fluorometric assay for the estimation of caspase 3, caspase 8 and caspase 9 activity, and Western blot for the determination of the levels of p53, nuclear factor-KB proteins and some of their target proteins. The A375 cells receiving no treatment served as the blank control group. Statistical analysis was carried out by t test. Results Within the investigated concentration and time ranges, paeonol significantly inhibited the proliferative activity of A375 cells in a concentration- and time- dependent manner. Compared with the blank control group, a significant increase was observed in the early apoptosis rate in A375 cells treated with paeonol of 1.25, 2.5 and 5 mmol/L for 24 hours （13.74% ±1.73%, 25.95%± 0.57% and 46.44% ± 0.81% vs. 3.11% ± 0.53%, P〈 0.05 or 0.01）, as well as in the activity of caspase 3, 8 and 9 in A375 cells treated with paeonol of 2.5 and 5 mmol/L for 24 hours （P 〈 0.05 or 0.01 ）. After 24-hour treatment, the protein levels of p53 and Bax were elevated, but those of nuclear factor-κB, Bcl-2 and Bcl-XL were decreased in A375 cells with the increase of paeonol concentration. Conclusions Paeonol can inhibit the proliferation but induce the apoptosis of A375 cells, and the apoptosis-inducing effect may be realized through intrinsic and extrinsic pathways by modulating nuclear factor-κB and p53 genes.

关 键 词：黑色素瘤 实验性 丹皮酚 细胞凋亡 细胞增殖 

分 类 号：R273[医药卫生—中西医结合]