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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(50、100、200、400及800nmol·L?1)诱导刺激不同时间,观察细胞凋亡的变化。通过Hochest/PI、AnnexinV-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;Westernblotting测定细胞色素c(Cyt-c)、Bcl-2、Bax、T-AKT及p-AKT的蛋白含量。结果显示:地塞米松(50～800nmol·L?1)作用48h后,可诱导β细胞凋亡;100nmol·L?1地塞米松处理72h后,caspase-3含量明显增多;不同浓度地塞米松处理48h,随着浓度的增加Cyt-c表达增多,Bax蛋白含量无明显变化,而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。
To observe the effect of dexamethasone on the apoptosis of MIN6 islet β cells and the expression of apoptosis-related protein and phosphorylation of AKT in mouse MIN6 cells, the mouse MIN6 islet β cells were selected as the study object, with different concentrations of dexamethasone (50,100 , 200,400 and 800 nmol·L -1, respectively). The changes of apoptosis were observed at different times. The rate of apoptosis was detected by Hochest / PI and Annexin V-FITC apoptosis detection kit. The caspase-3 content was detected by caspase-3 kit. The expressions of Cyt-c, Bcl-2, Bax and T -AKT and p-AKT protein content. The results showed that the apoptosis of β-cell was induced by dexamethasone (50 ~ 800nmol·L-1) for 48h, and the caspase-3 content increased significantly after treated with 100nmol·L-1 dexamethasone for 72h. Different concentrations of dexamethasone After treatment for 48h, the expression of Cyt-c increased with the increase of concentration, while the content of Bax did not change significantly, while the content of Bcl-2 protein decreased. There was no significant change in T-AKT expression, while p-AKT decreased with dexamethasone concentration. Therefore, dexamethasone can induce apoptosis of MIN6 islet β cells in mice, and its mechanism may be related to inhibition of AKT phosphorylation and down-regulation of Bcl-2 expression.