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（alveolar macrophages， AM）免疫功能基因水平及肿瘤细胞肺转移的影响尚未明确。在本研究中，我们研究了吸烟对小鼠肺泡巨噬细胞基因水平、免疫功能和肿瘤细胞肺转移的影响。雌性C57BL/6小鼠使用Hamburg II型吸烟机每天吸20支香烟（Coresta Monitor 2号），持续10天。10天后，处死小鼠。AM采用支气管肺泡灌洗法（BAL）获得。采用混合淋巴细胞反应（MLR）检测AM的抗原提呈活性。用氢乙胺和二氯荧光（DCFH）流式细胞术分析AM中活性氧的产生。流式细胞术分析AM表面ⅱ类抗原和B7-1抗原的表达。RT-PCR分析AM中IL- 1 β和TNF- α基因mRNA的表达。8-OHdG分析DNA基因的损伤情况。采用Lewis肺癌细胞（LL/2）检测肿瘤细胞的肺转移情况。与未吸烟小鼠相比，吸烟小鼠AM抗原提呈活性显著（p<0.01）降低。吸烟小鼠AM的O_2^-生成显著（p<0.01）增加，而两组AM的H_2O_2生成无显著差异。吸烟小鼠肺组织中DNA的8-OHdG水平升高。与未吸烟小鼠相比，吸烟小鼠AM中ⅱ类和B7-1抗原阳性细胞明显减少（p<0.01）。与未吸烟小鼠相比，吸烟小鼠AM中IL-1 β和TNF- α mRNA基因表达降低。与未吸烟小鼠相比，吸烟小鼠的LL/2肿瘤细胞向肺转移的速度加快。这些结果表明，吸烟小鼠AM的O_2^-（而非H_2O_2）可能损害AM的抗原提呈活性、ⅱ类和B7-1抗原的表达以及肺组织DNA基因。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