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本研究探讨葛根有效成分对恶性白血病可能的凋亡诱导作用及其分子机制。采用葛根总黄酮(flavonoids of puerarin,PR)处理人早幼粒细胞白血病(APL)细胞株NB4,用MTT法检测细胞增殖抑制率;FITC-Annexin V/PI双染法检测细胞凋亡率;实时定量PCR检测pml/rarα、bcl-2、survivin基因表达;Western blot检测JNK、p38MAPK、FasL及caspase相关酶的变化。结果发现,PR能明显抑制NB4细胞增殖,并诱导细胞凋亡。随着PR浓度的增加,pml/rarα、bcl-2及survivin基因在mRNA水平表达下调,JNK、FasL、caspase3及caspase8蛋白表达增加,与PR浓度呈正相关;PR联合三氧化二砷(arsenic trioxide,ATO)处理后上述作用更为明显。结论:PR诱导NB4细胞凋亡的机制可能与JNK相关信号分子的活化有关;PR联合ATO具有协同诱导NB4细胞凋亡的作用。
This study was aimed to investigate the possible apoptosis-inducing effect of puerariae officinalis on malignant leukemia and its molecular mechanism. The cell line NB4 of human promyelocytic leukemia (APL) was treated with flavonoids of puerarin (PR), the cell proliferation inhibition rate was detected by MTT assay, and the apoptosis rate was detected by FITC-Annexin V / PI double staining. The expressions of pml / rarα, bcl-2 and survivin were detected by quantitative PCR. The changes of JNK, p38MAPK, FasL and caspase related enzymes were detected by Western blot. The results showed that PR can significantly inhibit the proliferation of NB4 cells and induce apoptosis. The mRNA expression of pml / rarα, bcl-2 and survivin were down-regulated with the increase of PR concentration. The expression of JNK, FasL, caspase3 and caspase8 were increased and positively correlated with PR concentration. PR combined with arsenic trioxide (ATO) After the above effect is more obvious. CONCLUSION: The mechanism of PR-induced NB4 cell apoptosis may be related to the activation of JNK-related signaling molecules. PR combined with ATO may induce the apoptosis of NB4 cells.