微囊藻毒素MC-LR对原代肝细胞的影响

Study on the Effects of Microcystin-LR on Primary Cultured Hepatocyte

[目的 ]研究微囊藻毒素 (MC -LR)对原代培养的肝细胞增殖、去分化过程、细胞周期和凋亡的影响 ,探讨其促进肝细胞增殖的机制。 [方法 ]分离原代大鼠肝细胞 ,用 0、0 1、1 0或 10 0 μg/LMC -LR染毒 ,采用四唑盐比色试验(MTT)、图像分析技术和流式细胞术观察细胞生长情况。 [结果 ]微囊藻毒素可显著促进肝细胞生长和伸展 ,三个染毒组肝细胞生长促进率分别是对照组的 10 2 42 %、10 7 88%和 111 5 2 % ,染毒组肝细胞伸展提前 ,细胞平均面积显著增加 ,呈现出时间 剂量 效应关系 ;同时观察到典型的凋亡改变和G2 /M期阻滞 ,对照组和三个染毒组肝细胞凋亡率分别为6 47%、2 2 2 0 %、37 73%和 49 7%。 [结论 ]MC -LR通过促进细胞伸展诱导肝细胞去分化 ,使细胞进入增殖阶段 ,同时又能导致细胞阻滞 ,不能顺利进入下一个细胞周期的G0 期 。

To study the mechanism of the effects of microcystin-LR on hepatocytes proliferation,dedifferentiation,cell cycle and apoptosis. In this study,MTT test,image analysis and flow cytometry were applied to assess the cell proliferation,cell spreading,cell cycle and apoptosis of primary cultured hepatocytes exposed to 0,0.1,1.0 or 10.0 μg/L MC-LR. Cell proliferation and cell spreading were promoted in a time and dose dependent manner following exposure to microcystin-LR. The accelerate rates of cell proliferation in the three tested groups were 102.47%,107.88% and 111.52%,respectively,compared with that of control group. Cell spreading ahead of schedule was detected in the tested groups and the means of cell surface in the tested groups increased as well. Synchronously,typical apoptotic changes as well as synchronization at the G 2/M-phase were both detected in the tested groups,the apoptosis rates of control group and the tested groups were 6.47%,22.20%,37.73% and 49.7%,respectively. [Conclusion] The results suggested that microcystin could affect hepatocyte cycles,and induce cell dedifferentiation and proliferation as well as cycle block,which might lead to cell apoptosis.