Lee Jiann-Gwu, Wang Yan-Shiu, Chou Chin-Cheng
School of Veterinary Medicine, National Taiwan University, No. 1, Sec. 4, Roosevelt Rd., Taipei 106, Taiwan.
School of Veterinary Medicine, National Taiwan University, No. 1, Sec. 4, Roosevelt Rd., Taipei 106, Taiwan.
Toxicol In Vitro. 2014 Jun;28(4):562-70. doi: 10.1016/j.tiv.2014.01.005. Epub 2014 Jan 18.
This study aimed to evaluate the acrylamide (ACR)-induced apoptotic effects on rat primary astrocytes and three human astrocytoma-derived cell lines (U-1240 MG, U-87 MG, and U-251 MG). As determined through the MTT assay, treatment with 1 and 2 mM ACR for 24-72 h resulted in decreased cell viability in all cells. Decreases in cell viability could be blocked in all cells with the exception of U-251 MG cells by Z-DEVD FMK. ACR-induced dose-dependent apoptotic effects were also demonstrated by increases in the sub-G1 phase cell population in all cells. The decreased expressions of pro-caspase 3, 8, and 9 and the interruption of the mitochondrial membrane potential were observed in all cells. Exposure to 2 mM ACR for 48 h resulted in increased Bax/Bcl-2 ratios in primary astrocytes and U-87 MG cells, whereas the overexpression of Bcl-2 was observed in U-1240 MG and U-251 MG cells. The ACR-induced increases in the levels of p53 and pp53 in primary astrocytes could be attenuated by caffeine. These results suggest the existence of a common apoptotic pathway among all cell types and that U-87 MG cells may be a suitable substitute in vitro model for primary astrocytes in future studies on ACR-induced neurotoxicity.
本研究旨在评估丙烯酰胺（ACR）对大鼠原代星形胶质细胞和三种人星形细胞瘤衍生细胞系（U - 1240 MG、U - 87 MG和U - 251 MG）的凋亡诱导作用。通过MTT法测定，用1 mM和2 mM ACR处理24 - 72小时导致所有细胞的细胞活力下降。除U - 251 MG细胞外，Z - DEVD FMK可阻断所有细胞的细胞活力下降。所有细胞中G1期以下细胞群体的增加也证明了ACR诱导的剂量依赖性凋亡作用。在所有细胞中均观察到前半胱天冬酶3、8和9的表达降低以及线粒体膜电位的中断。暴露于2 mM ACR 48小时导致原代星形胶质细胞和U - 87 MG细胞中Bax/Bcl - 2比值增加，而在U - 1240 MG和U - 251 MG细胞中观察到Bcl - 2过表达。咖啡因可减弱ACR诱导的原代星形胶质细胞中p53和pp53水平的升高。这些结果表明所有细胞类型中存在共同的凋亡途径，并且在未来关于ACR诱导的神经毒性研究中，U - 87 MG细胞可能是原代星形胶质细胞合适的体外替代模型。
