论文部分内容阅读
目的:探讨肿瘤坏死因子相关凋亡诱导配体(TRAIL)联合小檗碱诱导Molt-4细胞凋亡作用及其对NF-κB/P65表达的影响。方法:采用四甲基偶氮唑(MTT)比色法测定TRAIL单独和联合应用小檗碱时对Molt-4细胞的生长抑制率;用流式细胞术和光镜细胞形态观察来检测凋亡;应用免疫印迹法检测单独和联合用药组细胞凋亡相关蛋白酶caspase3,caspase8及NF-κB/P65表达。结果:(1)MTT结果发现小檗碱可增加TRAIL对Molt-4细胞的生长抑制率,且呈时间和剂量依赖性(P<0.05)。(2)流式细胞术可以检测到凋亡,光镜细胞形态观察可见凋亡特异性形态改变。(3)Western blot结果显示(a)单独用TRAIL组及TRAIL联合小檗碱用药组caspase3,caspase8活化程度随TRAIL质量浓度依次增加,联合用药组活化作用更强。(b)单独用TRAIL组NF-κB/P65表达随剂量增加而增加。联合小檗碱用药组NF-κB/P65表达与单独用TRAIL组相比则明显受抑制。结论:(1)小檗碱可协同TRAIL诱导Molt-4细胞凋亡。(2)小檗碱协同TRAIL诱导Molt-4细胞凋亡的分子机制涉及抑制NF-κB/P65表达和caspase3,caspase8剪切活化。
Objective: To investigate the apoptosis of Molt-4 cells induced by tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and berberine and its effect on the expression of NF-κB / P65. Methods: MTT assay was used to determine the inhibitory rate of berberine alone or in combination with TRAIL on the growth of Molt-4 cells. Apoptosis was detected by flow cytometry and morphological observation with light microscopy. Western blotting was used to detect the expression of caspase3, caspase8 and NF-κB / P65 in single and combined groups. Results: (1) MTT results showed that berberine can increase the growth inhibition rate of Molt-4 cells by TRAIL in a time and dose-dependent manner (P <0.05). (2) Apoptosis could be detected by flow cytometry. The morphological changes of apoptotic cells were observed under light microscope. (3) Western blot results showed that: (a) The activation of caspase 3 and caspase 8 in TRAIL combined with berberine alone increased with the concentration of TRAIL, and the combination of TRAIL and berberine had stronger activation. (b) NF-κB / P65 expression increased with dose in TRAIL alone. The expression of NF-κB / P65 in combination berberine group was significantly inhibited compared with TRAIL group alone. Conclusion: (1) Berberine can cooperate with TRAIL to induce Molt-4 cell apoptosis. (2) The molecular mechanism of berberine in TRAIL-induced apoptosis in Molt-4 cells was involved in the inhibition of NF-κB / P65 expression and caspase3 and caspase8 cleavage activation.