Study of gene level for effect of smoking on alveolar macrophage function and lung metastasis of tumor.

在以往的研究中，吸烟对肺泡巨噬细胞(AM)免疫功能基因水平和肿瘤细胞肺转移的影响尚不清楚。本研究旨在探讨吸烟对小鼠肺泡巨噬细胞免疫功能基因水平及肿瘤细胞肺转移的影响。雌性C57BL/6小鼠在汉堡II型吸烟机上连续暴露10天，每天吸烟20支(Coresta Monitor No.2)。10天后处死小鼠。通过支气管肺泡灌洗(BAL)获得AM。混合淋巴细胞反应(MLR)检测AM的抗原提呈活性。用二氯荧光素(DCFH)和氢乙胺(DCFH)流式细胞仪分析AM中的活性氧产生情况。用流式细胞仪检测AM表面抗原(Class II和B7-1)的表达。逆转录聚合酶链式反应(RT-PCR)检测AM中IL-1β和肿瘤坏死因子-DNAm RNA的表达，8-OHdG.T-α法检测AM基因的损伤情况用Lewis肺癌细胞(LL/2)检测肿瘤细胞的肺转移情况。吸烟小鼠AM的抗原提呈活性显著低于非吸烟小鼠(p&lt；0.01)。吸烟小鼠AM的O2~-产生显著增加(P<0.01)，而H_2O_2的产生两组无明显差异。吸烟小鼠肺组织DNA 8-OHdG水平升高。吸烟小鼠AM中II类和B7-1类抗原阳性细胞较非吸烟小鼠显著减少(P<0.01)。吸烟小鼠肺泡巨噬细胞IL-1β和肿瘤坏死因子-α基因表达明显低于非吸烟小鼠。与未吸烟小鼠相比，吸烟小鼠可促进LL/2肿瘤细胞向肺转移。上述结果提示，吸烟小鼠肺泡巨噬细胞的抗原提呈活性、II类抗原和B7-1类抗原的表达均受到损伤，肺组织DNA基因也可能受到损伤，而H_2O_2的作用不明显。较少

In previous studies, the effects of smoking on the gene level of alveolar macrophages(AM) immunological functions and lung metastasis of tumor cells are not well defined. In the present study, we investigated the effects of smoking on the gene level of alveolar macrophages immunological functions and lung metastasis of tumor cells in mice. Female C57BL/6 mice were exposed to 20 cigarettes (Coresta Monitor No.2) / day during 10 days using Hamburg II smoking machine. After 10 days, mice were sacrificed. The AM were obtained by brochoalveolar lavage (BAL). The antigen presenting activity of AM was assayed by mixed lymphocyte reaction (MLR). The reactive oxygen species productions in AM were analyzed by flow cytometry using hydroethidine and dichlorofluorescen (DCFH). The expression of surface antigens (Class II and B7-1) of AM were analyzed by flow cytometry. The expressions of IL- 1 β and TNF- α mRNA of gene in AM were analyzed by RT-PCR.The damages of DNA genes were analyzed by 8-OHdG.T … More he lung metastases of tumor cells were assayed by using Lewis lung carcinoma cells(LL/2). The antigen presenting activity of AM was significantly (p<0.01) decreased in smoked mice compared with non-smoked mice. The O_2^- production of AM was significantly (p<0.01) increased in smoked mice, whereas the H_2O_2 production of AM was not difference in both groups. The 8-OHdG levels of DNA in lung tissue were increased in smoked mice. The Class II and B7-1 antigen positive cells in AM were significantly(p<0.01) decreased in smoked mice compared with non-smoked mice. IL-1 β and TNF- α mRNA genes expressions in AM were decrease in smoked mice as compared to non-smoked mice. The metastases to lung of LL/2 tumor cells were promoted in smoked mice compared with non-smoked mice. These results suggest that the antigen presenting activity, expressions of Class II and B7-1 antigens of the AM may be impaired and DNA gene in lung tissue also may be damaged by O_2^- but not H_2O_2 from AM in smoked mice. Less