续断皂苷抗白血病作用的机制探讨  

Study on the Mechanism of Akebia Saponin D for Leukemia

作　　者：周鹏[1] 马亮[1] 周珺[1] 国风[1] 

机构地区：[1]苏州大学附属第一医院中心实验室,江苏苏州215006

出　　处：《中国中西医结合杂志》2012年第1期84-88,共5页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：江苏省卫生厅面上项目(No.H200877);国家自然科学基金面上项目(No.81070405;No.81172433);教育部回国基金(No.K4122911)

摘　　要：目的探讨续断皂苷(akebia saponinD,ASD)抗白血病细胞及其作用机制。方法用不同剂量(10、30、50、100μg/mL)皂苷处理人急性白血病细胞株U937细胞和人早幼粒白血病细胞株HL-60细胞,采用MTT比色法观察ASD对白血病细胞的作用。以Annexin-Ⅴ染色法检测细胞凋亡并以PCR法检测凋亡相关基因的表达。采用Westernblot法检测P53蛋白的变化。采用Griess分光光度法检测样品中一氧化氮(NO)的代谢产物亚硝酸盐的含量。结果与未处理组相比,50μg/mLASD能明显抑制U937细胞和HL-60细胞的生长并呈剂量依赖性,同时伴随bcl-2mRNA表达的明显下调。Western blot检测结果显示,P53蛋白的表达水平随ASD作用而升高。另外,ASD可以促使U937细胞和HL-60细胞中NO的含量显著提高(P<0.05)。结论 ASD对U937细胞和HL-60细胞具有增殖抑制和诱导凋亡的作用,其机制与bcl-2基因的下调,p53蛋白上调,以及促进白血病细胞内NO含量提高相关。Objective To investigate the mechanism of akebia saponin D （ASD） for fight against leukemia cells. Methods The human acute leukemia cell strain U937 and human acute promyelocytic leukemia cell strain HL-60 were treated with ASD at different doses （10, 30, 50, and 100 tjg/mL）. The proliferation inhibition of ASD was observed using MTT assay. Apoptosis was detected by Annexin- staining. The expressions of cor- related genes were detected by PCR. Changes of p53 were detected using Western blot. The nitrite content, as the metabolite of nitric oxide （NO） was detected by Griess spectrophotometry. Results 50 iJg/mL ASD could ob- viously inhibit the growth of U937 and HL-60 cells in a dose-dependent manner, accompanied with the down-reg- ulation of bcl-2 mRNA expression. Results of Western blot showed that the P53 expression increased along with the dose of ASD. Besides, ASD could elevate the content of NO in U937 and HL-60 cells （P 〈0.05）. Conclusions ASD could inhibit the proliferation and induce the apoptosis of U937 and HL-60 cells. The mechanism might be correlated with down-regulating the bcl-2 expression, up-regulating the p53 expression, and increasing the NO content in U937 and HL-60 cells.

关 键 词：续断皂苷 白血病细胞 凋亡 P53 一氧化氮 

分 类 号：R285[医药卫生—中药学]