Yuan Fengjiao, Dong Ping, Wang Xiang, Fu Xiao, Dai Mingjun, Zhang Weiyun
Jiangsu Key Laboratory of Molecular Medicine, Medical School and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, China.
Exp Toxicol Pathol. 2013 Nov;65(7-8):1011-8. doi: 10.1016/j.etp.2013.02.004. Epub 2013 Mar 19.
Cigarette smoke exposure is associated with increased risk of different disorders. Immunological dysfunction especially in macrophages is one of important reasons in the initiation, progression and exacerbation of smoke-related pulmonary illnesses. However, it is still obscure how cigarette smoke impacts the vitality and functions of macrophages. In the present study, we examined the effects of cigarette smoke extract (CSE) on mouse Ana-1 macrophages and tried to elucidate the involved mechanism. The results showed CSE induced cell apoptosis accompanied by increased releasing of lactate dehydrogenase (LDH), mitochondrial injury and oxidative stress. It also inhibited anti-apoptosis protein Bcl-2 expression and promoted pro-apoptosis protein Bax and Bad expressions. Moreover, low-dose CSE increased nuclear NF-κB levels of macrophages; on the contrary, high-dose CSE or long-time treatment decreased it. These observations were in correspondence with changes of intracellular ROS level and antioxidant enzymes' activity. Furthermore, pretreatment with 10μM of NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC) for 1h significantly enhanced macrophage apoptosis. Taken together, these data implied that mitochondrial dysfunction and oxidative stress played important roles in the injury of Ana-1 cells caused by CSE, which was related to NF-κB pathway; an anti-apoptotic program played a dominant role at low doses/short-term exposure to CSE, whereas a pro-apoptotic program was initiated at high doses/long-term exposure.
接触香烟烟雾与患不同疾病的风险增加有关。免疫功能障碍，尤其是巨噬细胞的免疫功能障碍，是与烟雾相关的肺部疾病发生、发展和恶化的重要原因之一。然而，香烟烟雾如何影响巨噬细胞的活力和功能仍不清楚。在本研究中，我们检测了香烟烟雾提取物（CSE）对小鼠Ana-1巨噬细胞的影响，并试图阐明其中涉及的机制。结果显示，CSE诱导细胞凋亡，同时伴有乳酸脱氢酶（LDH）释放增加、线粒体损伤和氧化应激。它还抑制抗凋亡蛋白Bcl-2的表达，促进促凋亡蛋白Bax和Bad的表达。此外，低剂量CSE可增加巨噬细胞核NF-κB水平；相反，高剂量CSE或长时间处理则会降低其水平。这些观察结果与细胞内活性氧水平和抗氧化酶活性的变化一致。此外，用10μM的NF-κB抑制剂吡咯烷二硫代氨基甲酸盐（PDTC）预处理1小时可显著增强巨噬细胞凋亡。综上所述，这些数据表明线粒体功能障碍和氧化应激在CSE引起的Ana-1细胞损伤中起重要作用，这与NF-κB途径有关；在低剂量/短期接触CSE时，抗凋亡程序起主导作用，而在高剂量/长期接触时则启动促凋亡程序。
