红花多糖对人非小细胞肺癌A549细胞增殖和凋亡的影响

摘要

目的 探讨不同浓度的红花多糖(safflower polysaccharide,SPS)对人非小细胞肺癌A549细胞增殖和凋亡的影响.方法 根据台盼蓝染色法绘制细胞生长曲线及计算细胞活力;用不同浓度(0.04,0.08,0.16,0.32,0.64,1.28,2.56mg/mL)的红花多糖处理A549细胞24、48、72h后在倒置显微镜下观察细胞形态学改变;四甲基偶氮唑盐(MTT)法检测不同浓度(0.04,0.08,0.16,0.32,0.64,1.28,2.56mg/mL)的红花多糖对人非小细胞肺癌细胞体外生长的抑制作用,AnnexinV-FITC/PI荧光双标流式细胞术检测细胞凋亡率.结果 台盼蓝染色后发现在第2-6天细胞呈对数生长,在第3-5天细胞活力最好;不同浓度SPS处理24,48,72h后,各时间点均以0.64mg/mL的SPS抑制率最高;不同浓度SPS处理48h后,倒置显微镜下A549细胞表现为皱缩、变圆等细胞凋亡性;各浓度组A549细胞的凋亡率增加呈剂量依赖性,而1.28mg/mL组除外.结论 红花多糖能明显抑制人非小细胞肺癌A549细胞的增殖,诱导A549细胞的凋亡,在SPS浓度为0.64mg/mL最为显著.%Objective To investigate the potential anti-apoptotic effect of safflower polysaccharide on A549 cells of human non-small cell lung cancer. Methods Trypan blue staining was used to count the alive cells in order to draw a cellular growth curve and calculate cell viability. A549 cells were treated with different concentrations (0. 04, 0. 08, 0. 16, 0. 32, 0. 64, 1. 28 and 2. 56 mg/mL) of SPS for 24, 48 and 72 hours. Inverted Micro-scope was used to observe the morphological changes of A549 cells. Different doses of safflower polysaccharide (0. 04, 0. 08, 0. 16, 0. 32, 0. 64, 1. 28 and 2. 56 mg/mL) were added to A549 cells. The MTT assay was performed to re-veal the inhibitory effect on cell proliferation. Flow cytometry using annexin V-FITC/PI staining was employed to measure cell apoptosis. Results The Trypan blue staining showed the 2nd to 6th days was logarithmic growth phase and cell viability was the best from the 3rd to 5th days. After being treated by SPS for 24, 48 and 72 hours, the high-est inhibition rate was 0. 64mg/mL. After being treated by SPS for 48h, evident morphological changes including membrane shrinking, rounding shaping and budding to form apoptotic characters were observed. SPS treatment inhibi-ted the proliferation in a dose-dependent manner and the rate of cell apoptosis was increased also in a dose-dependent manner except 1. 28mg/mL. Conclusion SPS could inhibit the proliferation of A549 and enhance apoptosis of A549 especially in 0. 64mg/mL.