慢性砷暴露对人支气管上皮细胞超微结构的影响

发布时间：2018-12-16 09:37

【摘要】：目的研究长期亚砷酸钠诱导人支气管上皮细胞(HBE)恶性转化过程中线粒体及其他细胞器超微结构的改变,为砷致癌的机制研究提供动态细胞形态学改变基础资料。方法用1.0μmol/L亚砷酸钠连续染毒HBE细胞至43代,分别取正常和亚砷酸钠暴露1、8、15、22、29、36、43代HBE细胞,用流式细胞仪和生化法分别检测不同组别细胞内活性氧(ROS)、丙二醛(MDA)含量和超氧化物歧化酶(SOD)活力的变化,用透射电镜观察亚砷酸钠暴露15、29、43代和传代对照的HBE细胞内各细胞器超微结构的改变,用real-time PCR检测不同组别细胞内线粒体DNA(mtDNA)拷贝数的变化。结果与正常HBE细胞(0代)比较,亚砷酸钠染毒组22代及以前的ROS、MDA及SOD水平均呈先升高后下降的趋势;染毒29代及以后的ROS、MDA水平逐渐下降,SOD水平逐渐升高。透射电镜观察,与传代对照组细胞比较,染毒15代细胞的线粒体开始出现肿胀,嵴模糊,内质网扩张,且细胞mtDNA拷贝数增加;染毒29代及以后线粒体肿胀明显,嵴断裂,呈明显空泡状态,且有膜增殖、盘绕现象,mtDNA拷贝数先降低后升高,且均高于传代对照,差异有统计学意义(P0.05)。结论 1.0μmol/L亚砷酸钠在诱导HBE细胞恶性转化的过程中发生了细胞内氧化-抗氧化状态失衡,且随着染毒代数的增加,细胞线粒体DNA拷贝数增加明显,超微结构观察有细胞内线粒体肿胀、内质网扩张及膜增殖等变化。
[Abstract]:Objective to study the ultrastructural changes of mitochondria and other organelles during the malignant transformation of human bronchial epithelial cells (HBE) induced by sodium arsenite for a long time, and to provide basic data for the study of the mechanism of arsenic carcinogenesis. Methods HBE cells were continuously exposed to 1.0 渭 mol/L sodium arsenite to 43 passages, and normal and sodium arsenite exposed to 1 ~ 8152222 ~ 2936 ~ (43) HBE cells respectively. Flow cytometry and biochemical methods were used to detect reactive oxygen species (ROS),) in different groups of cells. The changes of malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in HBE cells were observed by transmission electron microscope. The changes of mitochondrial DNA (mtDNA) copy number in different groups of cells were detected by real-time PCR. Results compared with normal HBE cells (0 passage), the ROS,MDA and SOD levels in sodium arsenite treated group increased first and then decreased, ROS,MDA level decreased gradually and SOD level gradually increased after exposure to sodium arsenite. Compared with the control group, the mitochondria of the cells were swollen, cristae blurred, endoplasmic reticulum dilated, and the copy number of mtDNA increased. After 29 generations of exposure, mitochondria were swollen, cristae were broken and vacuolar, and there were membrane proliferation, coiling phenomenon, mtDNA copy number decreased first, then increased, and were higher than those of the control group, the difference was statistically significant (P0.05). Conclusion during the malignant transformation of HBE cells induced by 1.0 渭 mol/L sodium arsenite, the intracellular oxidation-antioxidation state imbalance occurred, and the DNA copy number of mitochondria increased significantly with the increase of exposure algebra. Ultrastructural changes of mitochondria swelling, endoplasmic reticulum dilatation and membrane proliferation were observed.
【作者单位】：苏州大学医学部公共卫生学院卫生毒理学教研室;
【基金】：国家自然科学基金(81573173) 高等学校博士学科点专项科研基金-博导类(20123201110012)
【分类号】：R114

【相似文献】