Chen Jingshu, Zhong Lei, Wu Jing, Ke Sui, Morpurgo Benjamin, Golovko Andrei, Ouyang Nengtai, Sun Yuxiang, Guo Shaodong, Tian Yanan
Texas A&M University.
Hunan Engineering Technology Research Center of Featured Aquatic Resources Utilization, Hunan Agriculture University.
J Vis Exp. 2018 Jun 25(136):57327. doi: 10.3791/57327.
Exposure to certain environmental chemicals in human and animals has been found to cause cellular damage of the pancreatic β cells which will lead to the development of type 2 diabetes mellitus (T2DM). Although the mechanisms for the chemical-induced β cell damage were unclear and likely to be complex, one recurring finding is that these chemicals induce oxidative stress leading to the generation of excessive reactive oxygen species (ROS) which induce damage to the β cell. To identify potential diabetogenic environmental chemicals, we isolated pancreatic islet cells from C57BL/6 mice and cultured islet cells in 96-well cell culture plates; then, the islet cells were dosed with chemicals and the ROS generation was detected by 2',7'-dichlorofluorescein (DCFH-DA) fluorescent dye. Using this method, we found that bisphenol A (BPA), Benzo[a]pyrene (BaP), and polychlorinated biphenyls (PCBs), could induce high levels of ROS, suggesting that they may potentially induce damage in islet cells. This method should be useful for screening diabetogenic xenobiotics. In addition, the cultured islet cells may also be adapted for in vitro analysis of chemical-induced toxicity in pancreatic cells.
已发现人类和动物接触某些环境化学物质会导致胰腺β细胞的细胞损伤，进而引发2型糖尿病（T2DM）。尽管化学物质诱导β细胞损伤的机制尚不清楚且可能很复杂，但一个反复出现的发现是，这些化学物质会诱导氧化应激，导致产生过量的活性氧（ROS），从而对β细胞造成损伤。为了识别潜在的致糖尿病环境化学物质，我们从C57BL/6小鼠中分离出胰岛细胞，并将其培养在96孔细胞培养板中；然后，用化学物质处理胰岛细胞，并使用2',7'-二氯荧光素（DCFH-DA）荧光染料检测ROS的产生。使用这种方法，我们发现双酚A（BPA）、苯并[a]芘（BaP）和多氯联苯（PCBs）可诱导高水平的ROS，表明它们可能会对胰岛细胞造成潜在损伤。这种方法应有助于筛选致糖尿病的外源性物质。此外，培养的胰岛细胞也可用于体外分析化学物质对胰腺细胞的毒性。
