Abella Vanessa, Santoro Anna, Scotece Morena, Conde Javier, López-López Verónica, Lazzaro Veronica, Gómez-Reino Juan Jesus, Meli Rosaria, Gualillo Oreste
SERGAS, Research Laboratory 9, NEIRID Lab (Neuroendocrine Interactions in Rheumatology and Inflammatory Diseases), Institute of Medical Research (IDIS), Santiago University Clinical Hospital, 15706 Santiago de Compostela, Spain; Departamento de Bioloxía Celular e Molecular, Facultade de Ciencias, Universidade da Coruña (UDC), Campus de A Coruña, 15071 A Coruña, Spain.
University of Naples Federico II, Dept. of Pharmacy, 80138 Naples, Italy.
Toxicol Lett. 2015 Apr 2;234(1):13-9. doi: 10.1016/j.toxlet.2015.02.001. Epub 2015 Feb 7.
Environmental pollutants are known to have adverse effects on human health. However, the link between chemical exposure and osteoarthritis remains little investigated. This study sought to assess in vitro the effect of several non-dioxin-like polychlorinated biphenyls (NDL-PCBs) on chondrocytes viability and apoptosis induction. Murine chondrogenic ATDC-5 cell line and human T/C-28a2 immortalized chondrocytes were exposed to NDL-PCBs 101, 153 and 180. Cell viability was examined using MTT assay. Necrosis was evaluated by LDH assay. Expression of apoptotic related proteins, such as caspase-3, Bcl-2 and Bax was assessed by Western blot analysis. Finally, oxidative stress was evaluated by malondialdehyde (MDA) assay and the Oxidative Stress Index. In vitro exposure to NDL-PCBs caused strong reduction of cell viability in a concentration-dependent manner. Data from LDH assay showed cellular necrosis induction. Caspase-3 activation, as well as, altered Bcl2/Bax ratio and p38 MAP-kinase phosphorylation also suggested apoptosis induction. Finally, MDA levels and Oxidative Stress Index revealed that PCBs drive chondrocyte death via increase of oxidative stress. The viability of murine and human chondrocytes was reduced in presence of PCBs. The activity of PCBs on cell viability is likely to be mediated by complex alterations involving regulation mechanisms of apoptosis, necrosis and oxidative stress.
已知环境污染物会对人类健康产生不利影响。然而，化学物质暴露与骨关节炎之间的联系仍鲜有人研究。本研究旨在体外评估几种非二噁英类多氯联苯（NDL-PCBs）对软骨细胞活力和凋亡诱导的影响。将小鼠软骨生成ATDC-5细胞系和人T/C-28a2永生化软骨细胞暴露于NDL-PCBs 101、153和180。使用MTT法检测细胞活力。通过乳酸脱氢酶（LDH）法评估细胞坏死情况。通过蛋白质免疫印迹分析评估凋亡相关蛋白如半胱天冬酶-3、Bcl-2和Bax的表达。最后，通过丙二醛（MDA）测定和氧化应激指数评估氧化应激。体外暴露于NDL-PCBs以浓度依赖性方式导致细胞活力显著降低。LDH测定数据显示诱导了细胞坏死。半胱天冬酶-3的激活以及Bcl2/Bax比值的改变和p38丝裂原活化蛋白激酶的磷酸化也表明诱导了凋亡。最后，MDA水平和氧化应激指数表明多氯联苯通过增加氧化应激导致软骨细胞死亡。在多氯联苯存在的情况下，小鼠和人类软骨细胞的活力降低。多氯联苯对细胞活力的作用可能是由涉及凋亡、坏死和氧化应激调节机制的复杂改变介导的。
