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[摘要]
目的 探讨人参多糖诱导白血病K562细胞凋亡的机制。方法 将对数生长期K562细胞分成对照组和人参多糖组,对照组细胞常规培养,人参多糖组细胞培养体系中加入人参多糖0.4 g/L。各组细胞培养48 h后,流式细胞术和Hoechst 33258染色法检测K562细胞凋亡情况;RT-PCR检测细胞p38、JNK基因表达;采用免疫荧光实验检测细胞中p-p38、p-JNK蛋白表达,测定Caspase-3的活性;Western blotting检测细胞中p38、JNK、p-p38、p-JNK、cleaved Caspase-3蛋白的变化。结果 与对照组比较,人参多糖组K562细胞凋亡率显著增加(P<0.05),出现明显的细胞核固缩现象,K562细胞p38 mRNA与JNK mRNA明显增多。免疫荧光检测显示,p-p38、p-JNK、cleaved Caspase-3表达明显增强且向胞核转移;Western blotting检测显示,人参多糖组K562细胞p38、JNK总蛋白无明显变化(P>0.05);p-p38、p-JNK、cleaved Caspase-3有增加的趋势(P<0.05)。结论 人参多糖能促进K562细胞凋亡,其作用可能是通过影响MAPK信号传导通路实现的。
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[Abstract]
Abstract: Objective To investigate the mechanisms of the ginseng polysaccharide (GPS) on the apoptosis of leukemia K562 cells. Methods The K562 cells at logarithmic growth phase were divided into control and GPS groups. The cells in the control group were normally treated and cells in GPS group were incubated with 0.4 g/L GPS. Flow cytometry and Hoechst 33258 staining were used to demonstrate the apoptotic changes in the two groups after incubation for 48 h. Gene expression of p38 and JNK were detected by RT-PCR. The immunofluorescence staining was used to detect the activation of Caspase-3 and expression of p-p38 and p-JNK protein. The changes of p38, JNK, p-p38, p-JNK, and cleaved Caspase-3 protein were detected by Western blotting. Results Compared with the control group, the apoptosis rate of K562 cells in GPS group was significantly increased (P < 0.05). After the treatment with GPS, chromatin condensation was observed when the cells were stained by Hochest 33258. The expression of p38 mRNA and JNK mRNA was obviously increased. The immunofluorescence staining results showed that the expression of p-p38, p-JNK, and cleaved Caspase-3 proteins was significantly increased in GPS group and obviously transferred to the nucleus. The Western blotting results showed that there was no significant change in total p38 and JNK protein (P < 0.05), but an increasing trend in p-p38, p-JNK, and cleaved Caspase-3 was observed (P < 0.05). Conclusion GPS could induce K562 cell apoptosis and the effect may be achieved through MAPK signal transduction pathway.
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[基金项目]
国家自然科学基金面上项目(81171929,31271368);重庆市教委基金项目(KJ110308)