【摘要】 目的观察三萜皂苷混合物ardisiacrispin(A+B)对人白血病HL-60细胞增殖的抑制作用,并对其可能的作用机制进行探讨。方法应用MTT法测定增殖抑制作用,采用流式细胞术(FACS)分析阻断细胞周期及诱导凋亡作用,采用PI荧光染色观察凋亡小体的存在,以Western blot分析凋亡相关蛋白表达变化。结果ardisiacrispin(A+B)可明显抑制HL-60细胞的增殖,具有浓度依赖性,作用48h的IC50值为4.2mg·L-1。ardisiacrispin(A+B)可阻断HL-60细胞于S期,在1.0～5.0mg·L-1范围内具有浓度依赖性,3.5mg.L-1ardisiacrispin(A+B)作用6、24和48h时,给药组中S期细胞比例分别为对照组的1.26、1.46和1.71倍。ardisia-crispin(A+B)可诱导HL-60细胞凋亡,在1.0～7.5mg.L-1浓度范围内具有浓度依赖性;此外,ardisiacrispin(A+B)可浓度依赖性地诱导PARP蛋白的裂解,2.5mg.L-1ardisi-acrispin(A+B)作用24h使裂解比例达到27.1%。结论Ardisiacrispin(A+B)可通过阻断细胞于S期、诱导凋亡来抑制HL-60细胞的增殖。更多还原

【Abstract】 Aim To study the inhibitory effect of ardisiacrispin(A+B) on human promyeloleukemic HL-60 cells and the underlying mechanism.Methods Cell growth inhibition in vitro was evaluated with MTT as- say. Ardisiacrispin (A+B)-induced cell cycle block and apoptosis were investigated using flow cytometry assay. Apoptitic bodies were detected by PI staining. Western blot was applied to determine modification of certain protein related to apoptosis. Results Ardisiacrispin (A+B) displayed dose dependent growth inhibition ability on HL-60 cells with the IC50 value of 4.2 mg·L-1 for 48 h. Ardisiacrispin(A+B) dose-dependently arrested HL-60 cells at the S phase of the cell cycle in the range of 1.0～5.0 mg·L-1,cells at S phase in treated group was 1.26、1.46 and 1.71 times of control group for 6、24 and 48 h of incubauion,respectively. Moreover, ardisiacrispin (A+B) induced dose-dependent apoptosis of HL-60 cells in the concentration of 1.0～7.5 mg·L-1.Furthermore,the PARP cleavage was detectable in a dose-dependent manner,2.5 mg·L-1 ardisiacrispin (A+B) generated a 27.1% of lysis fraction for 24 h of incubation. Conclusion Ardisiacrispin(A+B) exhibits prominent abilities to inhibit the proliferation of HL-60 cells by blocking the cell cycle at S phase and inducing apoptosis.更多还原