王林曦 王燕萍 刘小莺 刘亮 王丽静 陈闻佳 刘晓红 刘礼斌. 地塞米松诱导小鼠MIN6胰岛β细胞凋亡及其对AKT磷酸化的影响J. 药学学报, 2009,44(11): 1216-1220.
引用本文: 王林曦 王燕萍 刘小莺 刘亮 王丽静 陈闻佳 刘晓红 刘礼斌. 地塞米松诱导小鼠MIN6胰岛β细胞凋亡及其对AKT磷酸化的影响J. 药学学报, 2009,44(11): 1216-1220.
WANG Lin-Xi, WANG Yan-Ping, LIU Xiao-Ying, LIU Liang, WANG Li-Jing, CHEN Wen-Jia, LIU Xiao-Gong, LIU Li-Bin. Dexamethasone-induced apoptosis of murine MIN6 pancreatic β-cells and its effect on AKT phosphorylationJ. 药学学报, 2009,44(11): 1216-1220.
Citation: WANG Lin-Xi, WANG Yan-Ping, LIU Xiao-Ying, LIU Liang, WANG Li-Jing, CHEN Wen-Jia, LIU Xiao-Gong, LIU Li-Bin. Dexamethasone-induced apoptosis of murine MIN6 pancreatic β-cells and its effect on AKT phosphorylationJ. 药学学报, 2009,44(11): 1216-1220.

地塞米松诱导小鼠MIN6胰岛β细胞凋亡及其对AKT磷酸化的影响

Dexamethasone-induced apoptosis of murine MIN6 pancreatic β-cells and its effect on AKT phosphorylation

  • 摘要:

    为观察地塞米松对小鼠MIN6胰岛β细胞凋亡及其对凋亡相关蛋白表达和AKT磷酸化的影响, 选用小鼠MIN6胰岛β细胞为研究对象, 用不同浓度地塞米松 (50100200400800 nmol·L−1) 诱导刺激不同时间, 观察细胞凋亡的变化。通过Hochest/PIAnnexin V-FITC细胞凋亡检测试剂盒观测细胞凋亡率; caspase-3检测试剂盒测定caspase-3含量; Western blotting测定细胞色素c (Cyt-c)Bcl-2BaxT-AKTp-AKT的蛋白含量。结果显示: 地塞米松 (50800 nmol·L−1) 作用48 h, 可诱导β细胞凋亡; 100 nmol·L−1地塞米松处理   72 h, caspase-3含量明显增多; 不同浓度地塞米松处理48 h, 随着浓度的增加Cyt-c表达增多, Bax蛋白含量无明显变化, Bcl-2蛋白含量有所降低。T-AKT表达无明显变化, p-AKT随地塞米松浓度增加表达减少。因此, 地塞米松能够引起小鼠MIN6胰岛β细胞凋亡, 其机制可能与抑制AKT磷酸化、下调Bcl-2表达有关。

     

    Abstract:

    This study is to investigate the effect of dexamethasone on cell apoptosis of murine MIN6    pancreatic β-cells, and to investigate the mechanism of dexamethasone-dependent cell apoptosis.  The cell apoptosis model was established by choosing the murine MIN6 pancreatic β-cells, which was cultured in vitro and induced by dexamethasone.  The morphology of the cell apoptosis was observed through fluorescence   microscopic analysis after Hochest/PI staining and flow cytometric assay after Annexin-V/PI staining.  The  expression of caspase-3 was detected with caspase-3 activity assay kit.  The expressions of Cyt-c, Bcl-2,     Bax, AKT and p-AKT were observed with Western blotting.  The results indicated that after exposure to  dexamethasone at a concentration ranging from 50800 nmol·L−1 for 48 h, the percentage of cell apoptosis   was significantly increased with the concentration over 100 nmol·L−1 of dexamethasone; after exposure to  dexamethasone (100 nmol·L−1) for 72 h, the activity of caspase-3 increased significantly; after exposure to  dexamethasone at a concentration ranging from 50800 nmol·L−1 for 48 h, the expression of Cyt-c increased, Bcl-2 and AKT phosphorylation decreased while Bax and T-AKT remained unchanged.  It could be concluded that the effect of dexamethasone on murine MIN6 pancreatic β-cells apoptosis is significant.  The mechanism of dexamethasone-dependent cell apoptosis is probably related to down regulation of the Bcl-2 expression and    reduction of AKT phosphorylation.

     

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