二烯丙基二硫上调Beclin1 介导白血病K562 细胞自噬性死亡的研究

目的：通过二烯丙基二硫诱导白血病K562 细胞发生自噬性死亡，探讨其作用机制。方法：40 mg/LDADS 作用K562 细胞12 小时后，透射电镜观察K562 细胞超微结构，MDC 染色荧光显微镜观察自噬泡及流式细胞仪定量检测自噬率，RT-PCR 检测Beclin1mRNA 的表达水平。结果：DADS 作用后的K562 细胞后，透射电镜可观察到胞质内出现大量自噬体；MDC染色荧光显微镜观 察显示，K562 细胞胞浆中的自噬泡明显增多，而空白组与溶媒组胞浆中的自噬泡很少；流式细胞术定量测定空白对照组、溶媒对 照组、DADS药物组自噬率分别为(7.27± 5.60)%、(7.10± 5.13)%、(27.39± 6.51)%（P<0.05）；空白对照组为0.658± 0.007，溶媒对 照组为0.671± 0.012，两者的Beclin1mRNA 的表达强度无明显差异（P>0.05），DADS 药物组为0.911± 0.008，高于对照组（P<0. 05）。结论：二烯丙基二硫可诱导白血病k562 细胞发生自噬性死亡，其机制可能与Beclin1 的上调有关。

Diallyl Disulfide Raised Beclin1 Mediated Autophagy in Leukemia K562 Cell

Objective: To investigate the possibility of autophagic death of K562 cell induced by diallyl disulfide (DADS) and its possible mechanisms.Methods:K562 cells was treated by 40mg/L DADS for 12 hours. Its ultrastructure was observed by transmission election microscopy (TEM); Using monodansylcadaverin (MDC) staining, the autophagic vacuoles (AV) was observed by fluorescence microscope and the rates of autophagy was analyzed by flow cytometry; RT-PCR was employed to detect the expression levels of Beclin1mRNA.Results: After treatment with DADS, large amount of autophagosomes were observed in cytoplasm of K562 cell by TEM. Compared with blank reference group and the vehicle group, the autophagic vacuoles determined by MDC staining fluorescence microscope were increased significantly in DADS medication group. And the autophagy rate of blank reference group, vehicle reference group and DADS medication group, measured by means of flow cytometer, were (7.27± 5.60)%,(7.10± 5.13)%, (27.39± 6.51)%(P<0. 05) respectively. Higher Beclin1mRNA expression level (0.911± 0.008) was found in DADS medication group compared with blank reference group (0.658± 0.007) and vehicle reference group (0.671± 0.012), while no significant difference was observed between those contorl groups.Conclusion:DADS can induce autophagic death of leukemia K562 cell, the autophagic death may be related to the up-regulation of Beclin1 mRNA expression.