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目的探讨半合成紫草萘醌化合物SYUNZ-4对U937细胞的诱导凋亡作用及机制。方法锥虫蓝拒染实验和MTT法分析SYUNZ-4对U937细胞的生长抑制作用;DNA电泳、荧光显微镜观察细胞形态及流式细胞术检测细胞凋亡;Western blot法分析凋亡相关蛋白表达的变化。结果SYUNZ-4抑制U937细胞生长并呈时间和浓度依赖性,48 h的半数抑制浓度(IC50)为3．62μg／ml;经SYUNZ-4处理的U937细胞可见DNA梯形条带、细胞核浓缩和凋亡小体;6μg／ml SYUNZ-4处理U937细胞6,12,24 h,细胞凋亡率分别为(12．2±6．5)％、(25．3±11．4)％和(56．2±6．5)％,对照组凋亡率为(2．1±0．8)％(P<0．05);SYUNZ-4可活化U937细胞中的caspase-3、caspase-8、caspase-9和PARP,上调Bax、P21、P27、Fas和FasL的表达,下调Bcl-xL水平,对Bcl-2、BID和P53表达无影响。结论SYUNZ-4在体外可诱导U937细胞凋亡。caspase家族蛋白及其相关信号分子参与了凋亡的调节。
Objective To investigate the apoptosis-inducing effect and mechanism of semi-synthetic comfrey herb SYUNZ-4 on U937 cells. Methods The trypan blue exclusion test and MTT assay were used to analyze the growth inhibitory effect of SYUNZ-4 on U937 cells. DNA was electrophoresed and observed under fluorescence microscope. Flow cytometry was used to detect apoptosis. Western blot was used to analyze the expression of apoptosis-related proteins. Variety. Results SYUNZ-4 inhibited the growth of U937 cells in a time- and concentration-dependent manner. The IC50 of 48 h was 3.62 μg/ml. DNA ladders, nucleus condensation, and detachment were observed in U937 cells treated with SYUNZ-4. Apoptosis rate was (12.2±6.5)%, (25.3±11.4)%, and (56) for cells treated with 6 μg/ml SYUNZ-4 at 6, 12, and 24 h respectively. (2 ± 6.5)%, the apoptosis rate of the control group was (2.1 ± 0.8)% (P <0.05); SYUNZ-4 could activate caspase-3, caspase-8 in U937 cells, Caspase-9 and PARP up-regulated the expression of Bax, P21, P27, Fas, and FasL and down-regulated the Bcl-xL levels, but had no effect on the expression of Bcl-2, BID, and P53. Conclusion SYUNZ-4 can induce apoptosis of U937 cells in vitro. The caspase family proteins and their related signaling molecules are involved in the regulation of apoptosis.