小柴胡汤对HL-60白血病细胞株体外分化诱导作用探究.docVIP

小柴胡汤对HL-60白血病细胞株体外分化诱导作用探究 　【摘要】 　　目的观察小柴胡汤对HL-60白血病细胞的体外分化诱导作用，并初步探讨其作用机制。方法通过药物作用后观察细胞形态学的变化，细胞吞噬功能及硝基蓝四氮唑（NBT）还原能力的改变；流式测定细胞周期改变和免疫组化检测抑癌基因P53的表达变化初步揭示其作用机制。结果小柴胡汤能使HL-60细胞显著向成熟粒系细胞分化而且吞噬功能、NBT还原能力出现增强的趋势，以中、高剂量组最为显著(Plt;0.05)；并使 HL-60的细胞周期阻滞于G0/G1期,S期细胞数减少，抑癌基因P53表达明显升高(Plt;0.05)。结论小柴胡汤可诱导HL-60细胞向粒系成熟分化；其作用机制可能与阻滞细胞于G0/G1期，并且上调抑癌基因P53的表达有关。 【关键词】 小柴胡汤 HL-60 分化诱导 　　 　　Abstract：ObjectiveTo observe the effects of HL-60 cell differentiation induced by Xiaocaihu decoction and to discuss its mechanism.MethodsThe cells differentiation was observed on some changes of cytomorphology, the cell swallow function and NBT reducing ability. According to the changes of cell cycle by flow cytometry and anti-oncogene P53 expression by immunity histochemistry, the mechanism was revealed preliminarily.ResultsCompared with the blank control group ,the cell differentiation was induced into mature granulocyte morphologically and HL-60 cell phagocytosis and NBT reduction ability were enhanced markedly in middle-dose and high-dose Xiaochaihu decoction groups(Plt;0.05). Compared with the blank control group, HL－60 cell cycle was blocked into G0/G1 phase and the cell amount in S phase was decreased in middle-dose and high-dose Xiaochaihu decoction groups and P53 expression was enhanced significantly (Plt;0.05) in high-does Xiaocaihu decoction group.ConclusionHL－60 cells can be induced differentiation into mature granulocyte by Xiaocaihu decoction and the mechanism may be related with blocking cells into G0/G1 phase and increasing P53 expression. 　　Key words：Xaochaihu decoction； HL-60； Differentiation induction 　　长期以来“一旦成为癌细胞永远就是癌细胞”的概念一直统治着肿瘤学领域，但随着科技的不断深入，目前肿瘤逆转研究已成为肿瘤生物学的一个热门课题，对传统中医中的代表方剂小柴胡汤现代研究发现其具有广泛和多方面的抗癌作用［1～4］。 　　为了科学全面揭示此方的抗癌机理以及开发出新的诱导分化剂，我们选择对小柴胡汤诱导HL-60肿瘤细胞分化及其作用机制进行初步探索，以不断完善这首著名方剂抗癌作用的研究和挖掘它诱导肿瘤细胞分化的潜力。 　　1 材料与仪器 　　1.1 细胞株人早幼粒白血病HL-60细胞株由中国医学科学院药物研究所赠送。 　　1.2 药品小柴胡汤水煎液，配方：柴胡12 g，黄芩4.5 g，半夏3.75 g，生姜4.5 g，人参4.5 g，大枣4 g，炙甘草4.5 g，均购于北京同仁堂成都分店；剂量按原方比例确定；药液煎煮、离心取上清液，经40～60℃