喹乙醇诱导HepG2 细胞自噬的实验研究

目的： 研究喹乙醇对HepG2细胞自噬的影响。方法：分别用不同浓度 (0、200、400、800 μg/mL)喹乙醇染毒HepG2细胞24 h后，进行单丹磺酰尸胺 (monodansylcadaverine，MDC)染色，分别通过荧光显微镜和流式细胞仪检测细胞自噬发生情况。另外，分别用不同浓度喹乙醇 (0、200、400、800 μg/mL)染毒HepG2细胞24 h和400 μg/mL喹乙醇染毒HepG2细胞不同时间 (0、1、3、6、12、24 h)后，采用Western blot方法检测自噬相关蛋白LC3-Ⅱ和Beclin 1的表达。结果：随着喹乙醇染毒剂量增加，MDC阳性细胞的荧光强度及细胞内MDC荧光颗粒数目明显增加。流式结果显示，与对照组相比，200、400和800 μg/mL染毒组MDC阳性细胞百分比均显著增加 (P<0.05或P<0.01)。随着喹乙醇染毒浓度和时间的增加，LC3-Ⅱ和Beclin 1表达量均明显增加，其中400 μg/mL 喹乙醇染毒细胞24 h组与对照组相比，LC3-Ⅱ和Beclin 1的表达量均明显上调 (P<0.01)。结论：喹乙醇诱导了HepG2细胞的自噬反应。

Autophagic process induced by olaquindox in HepG2 cells

OBJECTIVE: To investigate the effect of olaquindox on autophagy in human hepatoma G2 (HepG2) cells. METHODS：The HepG2 cells were treated with different concentrations (0，200，400，800 μg/mL) of olaquindox for 24 h. Then autophagy was analyzed by fluorescence microscope and flow cytometer following staining with monodansylcadaverine (MDC). HepG2 cells were treated with different concentrations (0，200，400，800 μg/mL) of olaquindox for 24 h or with 400 μg/mL olaquindox for different periods (0，1，3，6，12，24 h). Then the expressions of LC3-Ⅱ and Beclin 1 were determined by Western blot. RESULTS：Olaquindox-treated cells exhibited higher fluorescent density and more MDC-labeled particles in HepG2 cells compared with the control group. The percentage of MDC-positive cells was obviously increased after treatment with different concentrations (200，400，800 μg/mL) of olaquindox for 24 h. (P<0.05 or P<0.01). The expressions of LC3-Ⅱ and Beclin 1 increased with different concentrations and different time points of olaquindox treatment. Moreover，compared with the control group，the expression of LC3-Ⅱ and Beclin 1 were increased significantly with treatment of 400 μg/mL olaquindox for 24 h (P<0.01). CONCLUSION： Olaquindox enhanced the autophagic process in HepG2 cells.