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

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(P0.05,P0.01);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(P0.05);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(P0.05,P0.01)。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。

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

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-3 protein 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.