慢性砷暴露对人支气管上皮细胞超微结构的影响  

Chronic arsenic exposure on the ultratructure of human bronchial epithelial cells

作　　者：王小娟[1] 杨旭[1] 王大朋[1] 许熙国[1] 马园[1] 朱震[1] 邓晗依 李春春[1] 安艳 

机构地区：[1]苏州大学医学部公共卫生学院卫生毒理学教研室,江苏苏州215123

出　　处：《环境与健康杂志》2017年第8期662-665,共4页Journal of Environment and Health

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)

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(P<0.05)。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。Objective To observe the mitochondria and other organelles superfine institutions changes in the process of malignant transformation of human bronchial epithelial （HBE） cells induced by sodium arsenite. Methods HBE cells were continuously treated by 1.0 μmol/L sodium arsenite for 43 passages. 1, 8, 15, 22, 29, 36, 43 passages of HBE cells were selected as the research objects, the flow cytometry was used to detect the levels of reactive oxygen species （ROS）. MDA content was detected with thibabituric acid （TBA） test. The vitality unite of superoxide dismutase （SOD） was measured with total superoxide dismutase assay kit. The organelles uhrastructure in the 15, 29, 43 passages of HBE cells and the control group were observed by transmission electron microscopy （TEM）. The mtDNA copy numbers of the above groups were determined by real-time PCR. Results Compared with the control group, the levels of ROS, MDA and SOD increased and then decreased with the increase of passages before passage 22; After passage 29, however, the levels of ROS and MDA were declined to a much lower level gradually, SOD activity increased gradually. Compared with the control group, observation by transmission electron microscope revealed that there was a little swelling of mitochondria and indistinct cristae, the mtDNA copy number was increased before passage 15; After 29 passage, there were mitochondria swelling, fractured cristae, vacuolization, and membrane proliferation and coil, mtDNA copy number were reduced first and then rose, and also both higher than the control group. Conclusion There＇ is a disturbance of oxidation antioxidation mechanism during the malignant transformation of HBE cells induced by sodium arsenite, and with the increase of infected passages, mtDNA copy numbers change obviously, and the changes in the mitochondria are apparent, including swelling mitochondfia, endoplasmic reticulum expansion, membrane proliferation and so on.

关 键 词：亚砷酸钠 人支气管上皮细胞 线粒体 超微结构 

分 类 号：R994.6[医药卫生—毒理学]