木香挥发油诱导人白血病细胞MV4-11凋亡及其作用机制的探讨  

Essential Oil from Aucklandiae Radix Induces Apoptosis and Potential Mechanism in MV4-11 Leukemia Cells

作　　者：杨雨婷[1] 何育霖[1] 何贝轩 熊亮[1] 曹治兴[1] 彭成[1] 

机构地区：[1]成都中医药大学药学院四川省中药资源系统研究与开发利用重点实验室--省部共建国家重点实验室培育基地,成都611137

出　　处：《中国实验方剂学杂志》2016年第19期95-99,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：四川省杰出青年项目(2015JQ0030Q)

摘　　要：目的:以人急性髓性白血病MV4-11细胞为研究对象,探讨木香挥发油对MV4-11细胞增殖与凋亡抑制作用及其作用机制。方法:以质量浓度分别为3,6,12,25,50,100 mg·L^(-1)的木香挥发油作用于人急性髓性白血病MV4-11细胞不同时间,另设空白组,采用噻唑蓝(MTT)法检测木香挥发油对其增殖抑制作用,Hoechst 33258染色法观察细胞凋亡的形态学变化,Annexin V-FITC,PI双染及PI单染法应用流式细胞术检测细胞凋亡和周期,蛋白质免疫印迹(Western blot)检测蛋白激酶B(AKT),半胱氨酸天冬氨酸蛋白酶3(Caspase-3)凋亡蛋白的表达。结果:木香挥发油对MV4-11细胞的半抑制浓度(IC50)为13.33 mg·L^(-1);木香挥发油作用于MV4-11细胞24 h后可使细胞核皱缩,染色质凝聚,形成明显的凋亡小体;与空白组比较,木香挥发油G0/G1期和G2/M期细胞明显减少,S期细胞明显增多(P<0.05,P<0.01);与空白组比较,12,25,50,100,150 mg·L^(-1)木香挥发油能诱导MV4-11细胞凋亡(P<0.05);与空白组比较,25,50,100,150 mg·L^(-1)木香挥发油可抑制AKT的磷酸化,并促进Caspase-3蛋白的降解(P<0.05,P<0.01)。结论:木香挥发油能抑制人白血病细胞MV4-11细胞的增殖,其作用机制可能与抑制AKT活性进而诱导细胞凋亡效应有关。Objective： To investigate the effect of essential oil from Aucklandiae Radix on proliferation and apoptosis inhibition of MV4-11 leukemia cells and discuss its action mechanism. Method： Human leukemia cell line MV4-11 was treated with the essential oil from Aucklandiae Radix（ 3,6,12,25,50,100 mg·L-1） for different time,and another blank group was set up. Cell viability was estimated by using MTT assay; the apoptosis morphology changes were observed by Hoechst 33258 staining; flow cytometry was used on Annexin V-FITC /propidium iodide staining to detect the cell cycle and the rate of apoptosis; Western blot was used to detect AKT and Caspase-3 protein expression. Result： The essential oil from Aucklandiae Radix had IC50 value of 13. 33 mg·L-1; various nuclear changes such as nuclear shrinkage,chromatin condensation and obvious apoptotic bodies were observed after MV4-11 cell line was treated with essential oil from Aucklandiae Radix for 24 h. As compared with the blank group,the cells in G0/ G1 phase were significantly reduced and the cells in S phase were significantly increased with the increase from essential oil from Aucklandiae Radix oil dosage（ P 〈 0. 05,P 〈 0. 01）. 12,25,50,100,150 mg·L-1 essential oil from Aucklandiae Radix induced apoptosis of MV4-11（ P 〈 0. 05）; 25,50,100,150 mg·L-1 essential oil from Aucklandiae Radix inhibited AKT phosphorylation and promoted Caspase-3protein degradation（ P 〈 0. 05,P 〈 0. 01）. Conclusion： The essential oil from Aucklandiae Radix can inhibit the proliferation of MV4-11 cells in vitro,and the mechanism may be correlated with inducing cell apoptosis by inhibiting AKT activity.

关 键 词：木香挥发油 凋亡 人白血病MV4-11细胞 增殖 蛋白激酶B 半胱氨酸天冬氨酸蛋白酶3 

分 类 号：R285.5[医药卫生—中药学]