香烟烟雾凝集物诱导人支气管上皮细胞IL-8表达的机制
摘要
吸烟是呼吸系统疾病的重要危险因素。香烟烟雾刺激正常人呼吸道上皮细胞,可诱导白细胞介素-8(Interleukin-8, IL-8)的表达增加。IL-8基因顺式调控元件内有包括核转录因子-κB(nuclear factor-κB, NF-κB)和CCAAT/增强子结合蛋白-β(CCAAT/enhancer binding protein-β, C/EBPβ)在内的多个核因子结合位点。但有关NF-κB和C/EBPβ在香烟烟雾诱导IL-8基因转录活化的细胞内信号传导途径中的作用尚不清楚。
目的
该研究用香烟烟雾凝集物(cigarette smoke extracts, CSE)刺激人支气管上皮BEAS-2B细胞,观察CSE对BEAS-2B细胞的损害作用和对细胞炎症因子IL-8表达的影响。利用IL-8促进子上的NF-κB, C/EBPβ结合位点突变细胞株(IL-8mNF-κB-BEAS-2B cells,IL-8mC/EBPβ-BEAS-2B cells),细胞中被激活的NF-κB, C/EBPβ不能与IL-8促进子结合。通过比较CSE刺激后,NF-κB, C/EBPβ结合位点突变BEAS-2B细胞与野生型BEAS-2B细胞(IL-8 WT BEAS-2B cells)中与IL-8促进子相连的荧光素酶(luciferase,fLCF)基因表达量的变化,观察NF-κB和C/EBPβ转录因子在CSE引起的人支气管上皮细胞IL-8表达变化中的作用。为进一步探讨吸烟导致呼吸系统炎症发生机制提供实验依据。
实验方法
1 CSE溶液的配制
CSE原液的制备参照Su Y的方法并改良,CSE原液用无血清RPMI-1640培养液稀释,配制好的CSE于30min之内用于实验。
2 MTT比色法测CSE溶液对BEAS-2B细胞的半数抑制浓度
用0%、2.5%、5.0%、7.5%、10.0%、12.5%的CSE溶液刺激BEAS-2B细胞,测定不同浓度的CSE对BEAS-2B细胞的抑制作用,并得出CSE溶液的半数抑制浓度。
3 CSE处理正常BEAS-2B细胞及IL-8基因mRNA表达水平检测
根据MTT测定的结果,分别以浓度0%、2.5%、5.0%、7.5%、10.0%的CSE作用正常BEAS-2B细胞,通过荧光定量PCR,分析不同浓度CSE处理后细胞IL-8 mRNA水平的变化。
4 CSE处理3种突变型BEAS-2B细胞及其fLCF mRNA表达水平检测
根据CSE处理正常BEAS-2B细胞后IL-8基因表达的变化,用7.5%CSE作用3种突变型BEAS-2B细胞,通过荧光定量PCR,分析CSE处理后,细胞fLCF mRNA水平的变化。
5荧光素酶活性测定
以试剂盒中报告基因细胞裂解液为空白对照,测定7.5%CSE作用的正常BEAS-2B细胞和3种突变型BEAS-2B细胞的荧光素酶活性。
结果
1 BEAS-2B细胞的形态学观察
CSE刺激支气管上皮细胞来源的BEAS-2B细胞,可导致BEAS-2B细胞形态改变。CSE处理后,细胞胞浆疏松,漂浮细胞增多。随着CSE浓度的增加,细胞受损情况也越严重。
2 7.5%CSE处理3种突变型BEAS-2B细胞,细胞绿色荧光的变化
3种突变型BEAS-2B细胞受到CSE作用后,荧光显微镜下观察到绿色荧光细胞减少,细胞回缩变圆。
3 MTT检测结果
CSE对BEAS-2B细胞的生长有显著的抑制作用,F=25.06,P<0.001。且随着药物浓度的增加,CSE对细胞的抑制作用增强,呈剂量-效应关系。CSE对BEAS-2B细胞的半数抑制浓度约为7.5%。
4 BEAS-2B细胞IL-8 mRNA在不同浓度CSE作用后的RT-PCR检测结果
CSE作用BEAS-2B细胞,可导致其炎症因子IL-8表达增加,单因素方差分析表达差异有统计学意义(F=1808.1,P<0.001)。低浓度(2.5%、5.0%和7.5%)的CSE刺激增加IL-8 mRNA的表达(P<0.001),且具有浓度依赖性增高；但是10.0%CSE处理组大量细胞死亡,其IL-8 mRNA表达量与无CSE处理组间差异无统计学意义(P>0.05)。
5 7.5%CSE作用3种突变型BEAS-2B细胞fLCF mRNA的RT-PCR检测结果
7.5%CSE作用后,IL-8mNF-κB-BEAS-2B细胞和IL-8mC/EBPβ-BEAS-2B细胞的fLCF mRNA表达量都较IL-8 WT BEAS-2B细胞低,且IL-8 WT BEAS-2B细胞fLCF mRNA表达量是IL-8mNF-κB- BEAS-2B细胞fLCF mRNA表达量的1.32倍,是IL-8mC/EBPβ- BEAS-2B细胞的fLCF mRNA表达量的1.54倍。
6荧光素酶活性测定
相同浓度和作用时间的CSE刺激,IL-8mNF-κB- BEAS-2B细胞和IL-8mC/EBPβ- BEAS-2B细胞的荧光素酶活性都较IL-8WT BEAS-2B细胞低,且差异有统计学意义(P<0.001)。
结论
NF-κB转录因子参与CSE诱导的IL-8基因转录活化的细胞内信号传导过程。该研究首次证实C/EBPβ转录因子参与CSE诱导的IL-8基因转录活化。提不NF-κB和C/EBPβ转录因子通过参与炎症因子IL-8表达,促进吸烟导致的呼吸系统炎症反应。
Cigarette smoke mediated oxidative stress and inflammatory events in the bronchial epithelium are important processes in the pathogenesis of smoking related pulmonary diseases. Lots of harmful substances generated by smoking including free radicals and oxides can trigger pro-inflammatory cytokines, which are increased in the lungs of smokers. The airway epithelium is the primary target for any inhaled environmental agents and plays a critical role in the release of pro-inflammatory mediators. Previous in vivo findings have supported that cigarette smoke can induce pro-inflammatory cytokine (Interleukin-8, IL-8) release in smokers and in rodent lungs. However, the precise molecular mechanisms as how cigarette smoke generates signals for pro-inflammatory cytokine release, particularly in bronchial epithelium are not yet clearly understood.
Objective
Here we examined the acute effect of aqueous cigarette smoke extracts (CSE) in cultured normal human bronchial epithelial cell line (BEAS-2B), and detected the changes of the transcription of Interleukin-8. IL-8 is a multifunctional cytokine that has significant neutrophil chemoattractant and activating properties. It was reported that NF-κB and C/EBPβinvolve in IL-8 promoter transactivation as well as in its generation. Herein, we used three kinds of mutant BEAS-2B cells to determine effects of NF-κB and C/EBPβon the transcription of Interleukin-8 in cigarette smoke extract treated BEAS-2B cells. These three kinds of cells were transfected by luciferase plasmid and EGFP plasmid. Luciferase constructs under the control of the IL-8 promoter (IL-8 WT BEAS-2B cells), as well as mutants in the NF-κB site (IL-8mNFκB-BEAS-2B cells) or in the C/EBPβsite (IL-8mC/EBPβ-BEAS-2B cells).
Methods
1 Preparation of cigarette smoke extracts solution
CSE was prepared by using an improved modification of the method described by Su Y, and diluted by RPMI1640 medium without FBS.
2 Using MTT colorimetric methods to get IC50
BEAS-2B cells were treated by 0%,2.5%,5.0%,7.5%,10.0%,12.5%. CSE for 24h and added in MTT to get the inhibitions of BEAS-2B cells under the pression of different CSE concentrations. And then we got the Inhibitory Concentration 50(IC50).
3 CSE of different concentrations treated BEAS-2B cells, and we detected the transcription of IL-8 mRNA by fluorescent quantitative PCR
Based on the results of the MTT colorimetric methods, BEAS-2B cells were treated with 0%,2.5%,5.0%,7.5%,10.0%CSE for 24h for each experimental condition, total RNA was extracted and 1μg of the resulting RNA was then reverse transcribed to cDNA. And fluorescent quantitative PCR assay was used to detect the transcription of IL-8 mRNA.
4 7.5%CSE stimulated three kinds of mutant BEAS-2B cells, and we detected the transcription of fLCF mRNA by fluorescent quantitative PCR
Three kinds of mutant BEAS-2B cells were treated by the 7.5%CSE for 24h for each experimental condition, total RNA was extracted and 1μg of the resulting RNA was then reverse transcribed to cDNA. And fluorescent quantitative PCR assay was used to detect the transcription of fLCF mRNA.
5 Luciferase assays
7.5%CSE treated normal BEAS-2B cells and three kinds of mutant BEAS-2B cells for 24h. And then we measured the luciferase activity.
Results
1 Morphologic change of the BEAS-2B cells treated with CSE
After the stimulated by CSE, the shapes of BEAS-2B cells became irregular. There were more floating cells compared with cells without CSE. We observed a CSE dose dependant increase in cellular damage. And the cell damage became more seriously with the increase of CSE.
2 Changes of green fluorescent protein (EGFP) in three kinds of mutant BEAS-2B cells before and after CSE treated
CSE had an obvious cyto-inhibition. Cells with green fluorescent protein (EGFP) became fewer and round observed under the fluorescence microscope.
3 Results of the MTT colorimetric method
CSE had an obvious cyto-inhibition (F=25.06, P<0.001). With the increase of the concentration of CSE, cyto-inhibition became higher. There was a dose-effect relationship between CSE and the cyto-inhibition. And Inhibitory Concentration 50 of CSE was 7.5%.
4 Transcription of IL-8 mRNA in BEAS-2B cells treated with different concentrations of CSE detected by fluorescent quantitative PCR
BEAS-2B cells were stimulated with 2.5%,5.0%,7.5%and 10.0%CSE for 24h. IL-8 mRNA levels were up-regulated by CSE (2.5%,5.0%and 7.5%), (P<0.001)and had a CSE dose dependent increase. However CSE at a higher concentration (10.0%) made lots of cells dead and did not increase IL-8 mRNA level (P>0.05).
5 Transcription of fLCF mRNA in three kinds of mutant BEAS-2B cells treated with 7.5%CSE detected by fluorescent quantitative PCR
The transcriptions of fLCF mRNA of IL-8mNF-κB-and IL-8mC/EBPβ-BEAS-2B cells were lower than that of IL-8 WT BEAS-2B cells. The transcription of fLCF mRNA of IL-8 WT BEAS-2B cells was 1.32 times than that of IL-8mNF-κB-BEAS-2B cells and 1.54 times than that of IL-8mC/EBPβ-BEAS-2B cells.
6 Luciferase assays
7.5%CSE treated BEAS-2B cells and three kinds of mutant BEAS-2B cells for 24h. We found that the luciferase activities in IL-8mNF-κB-BEAS-2B cells and in IL-8mC/EBPp-BEAS-2B cells were lower than that in IL-8 WT BEAS-2B cells. And the discrepancy had a statistical significance (P< 0.05).
Conclusions
NF-κB regulated the release of Interleukin-8 in BEAS-2B cells treated by cigarette smoke extract. And this study showed for the first time that C/EBPβtranscription factor involved in the precise molecular mechanisms of the release of Interleukin-8 in BEAS-2B cells treated by cigarette smoke extract.
目的
该研究用香烟烟雾凝集物(cigarette smoke extracts, CSE)刺激人支气管上皮BEAS-2B细胞,观察CSE对BEAS-2B细胞的损害作用和对细胞炎症因子IL-8表达的影响。利用IL-8促进子上的NF-κB, C/EBPβ结合位点突变细胞株(IL-8mNF-κB-BEAS-2B cells,IL-8mC/EBPβ-BEAS-2B cells),细胞中被激活的NF-κB, C/EBPβ不能与IL-8促进子结合。通过比较CSE刺激后,NF-κB, C/EBPβ结合位点突变BEAS-2B细胞与野生型BEAS-2B细胞(IL-8 WT BEAS-2B cells)中与IL-8促进子相连的荧光素酶(luciferase,fLCF)基因表达量的变化,观察NF-κB和C/EBPβ转录因子在CSE引起的人支气管上皮细胞IL-8表达变化中的作用。为进一步探讨吸烟导致呼吸系统炎症发生机制提供实验依据。
实验方法
1 CSE溶液的配制
CSE原液的制备参照Su Y的方法并改良,CSE原液用无血清RPMI-1640培养液稀释,配制好的CSE于30min之内用于实验。
2 MTT比色法测CSE溶液对BEAS-2B细胞的半数抑制浓度
用0%、2.5%、5.0%、7.5%、10.0%、12.5%的CSE溶液刺激BEAS-2B细胞,测定不同浓度的CSE对BEAS-2B细胞的抑制作用,并得出CSE溶液的半数抑制浓度。
3 CSE处理正常BEAS-2B细胞及IL-8基因mRNA表达水平检测
根据MTT测定的结果,分别以浓度0%、2.5%、5.0%、7.5%、10.0%的CSE作用正常BEAS-2B细胞,通过荧光定量PCR,分析不同浓度CSE处理后细胞IL-8 mRNA水平的变化。
4 CSE处理3种突变型BEAS-2B细胞及其fLCF mRNA表达水平检测
根据CSE处理正常BEAS-2B细胞后IL-8基因表达的变化,用7.5%CSE作用3种突变型BEAS-2B细胞,通过荧光定量PCR,分析CSE处理后,细胞fLCF mRNA水平的变化。
5荧光素酶活性测定
以试剂盒中报告基因细胞裂解液为空白对照,测定7.5%CSE作用的正常BEAS-2B细胞和3种突变型BEAS-2B细胞的荧光素酶活性。
结果
1 BEAS-2B细胞的形态学观察
CSE刺激支气管上皮细胞来源的BEAS-2B细胞,可导致BEAS-2B细胞形态改变。CSE处理后,细胞胞浆疏松,漂浮细胞增多。随着CSE浓度的增加,细胞受损情况也越严重。
2 7.5%CSE处理3种突变型BEAS-2B细胞,细胞绿色荧光的变化
3种突变型BEAS-2B细胞受到CSE作用后,荧光显微镜下观察到绿色荧光细胞减少,细胞回缩变圆。
3 MTT检测结果
CSE对BEAS-2B细胞的生长有显著的抑制作用,F=25.06,P<0.001。且随着药物浓度的增加,CSE对细胞的抑制作用增强,呈剂量-效应关系。CSE对BEAS-2B细胞的半数抑制浓度约为7.5%。
4 BEAS-2B细胞IL-8 mRNA在不同浓度CSE作用后的RT-PCR检测结果
CSE作用BEAS-2B细胞,可导致其炎症因子IL-8表达增加,单因素方差分析表达差异有统计学意义(F=1808.1,P<0.001)。低浓度(2.5%、5.0%和7.5%)的CSE刺激增加IL-8 mRNA的表达(P<0.001),且具有浓度依赖性增高；但是10.0%CSE处理组大量细胞死亡,其IL-8 mRNA表达量与无CSE处理组间差异无统计学意义(P>0.05)。
5 7.5%CSE作用3种突变型BEAS-2B细胞fLCF mRNA的RT-PCR检测结果
7.5%CSE作用后,IL-8mNF-κB-BEAS-2B细胞和IL-8mC/EBPβ-BEAS-2B细胞的fLCF mRNA表达量都较IL-8 WT BEAS-2B细胞低,且IL-8 WT BEAS-2B细胞fLCF mRNA表达量是IL-8mNF-κB- BEAS-2B细胞fLCF mRNA表达量的1.32倍,是IL-8mC/EBPβ- BEAS-2B细胞的fLCF mRNA表达量的1.54倍。
6荧光素酶活性测定
相同浓度和作用时间的CSE刺激,IL-8mNF-κB- BEAS-2B细胞和IL-8mC/EBPβ- BEAS-2B细胞的荧光素酶活性都较IL-8WT BEAS-2B细胞低,且差异有统计学意义(P<0.001)。
结论
NF-κB转录因子参与CSE诱导的IL-8基因转录活化的细胞内信号传导过程。该研究首次证实C/EBPβ转录因子参与CSE诱导的IL-8基因转录活化。提不NF-κB和C/EBPβ转录因子通过参与炎症因子IL-8表达,促进吸烟导致的呼吸系统炎症反应。
Cigarette smoke mediated oxidative stress and inflammatory events in the bronchial epithelium are important processes in the pathogenesis of smoking related pulmonary diseases. Lots of harmful substances generated by smoking including free radicals and oxides can trigger pro-inflammatory cytokines, which are increased in the lungs of smokers. The airway epithelium is the primary target for any inhaled environmental agents and plays a critical role in the release of pro-inflammatory mediators. Previous in vivo findings have supported that cigarette smoke can induce pro-inflammatory cytokine (Interleukin-8, IL-8) release in smokers and in rodent lungs. However, the precise molecular mechanisms as how cigarette smoke generates signals for pro-inflammatory cytokine release, particularly in bronchial epithelium are not yet clearly understood.
Objective
Here we examined the acute effect of aqueous cigarette smoke extracts (CSE) in cultured normal human bronchial epithelial cell line (BEAS-2B), and detected the changes of the transcription of Interleukin-8. IL-8 is a multifunctional cytokine that has significant neutrophil chemoattractant and activating properties. It was reported that NF-κB and C/EBPβinvolve in IL-8 promoter transactivation as well as in its generation. Herein, we used three kinds of mutant BEAS-2B cells to determine effects of NF-κB and C/EBPβon the transcription of Interleukin-8 in cigarette smoke extract treated BEAS-2B cells. These three kinds of cells were transfected by luciferase plasmid and EGFP plasmid. Luciferase constructs under the control of the IL-8 promoter (IL-8 WT BEAS-2B cells), as well as mutants in the NF-κB site (IL-8mNFκB-BEAS-2B cells) or in the C/EBPβsite (IL-8mC/EBPβ-BEAS-2B cells).
Methods
1 Preparation of cigarette smoke extracts solution
CSE was prepared by using an improved modification of the method described by Su Y, and diluted by RPMI1640 medium without FBS.
2 Using MTT colorimetric methods to get IC50
BEAS-2B cells were treated by 0%,2.5%,5.0%,7.5%,10.0%,12.5%. CSE for 24h and added in MTT to get the inhibitions of BEAS-2B cells under the pression of different CSE concentrations. And then we got the Inhibitory Concentration 50(IC50).
3 CSE of different concentrations treated BEAS-2B cells, and we detected the transcription of IL-8 mRNA by fluorescent quantitative PCR
Based on the results of the MTT colorimetric methods, BEAS-2B cells were treated with 0%,2.5%,5.0%,7.5%,10.0%CSE for 24h for each experimental condition, total RNA was extracted and 1μg of the resulting RNA was then reverse transcribed to cDNA. And fluorescent quantitative PCR assay was used to detect the transcription of IL-8 mRNA.
4 7.5%CSE stimulated three kinds of mutant BEAS-2B cells, and we detected the transcription of fLCF mRNA by fluorescent quantitative PCR
Three kinds of mutant BEAS-2B cells were treated by the 7.5%CSE for 24h for each experimental condition, total RNA was extracted and 1μg of the resulting RNA was then reverse transcribed to cDNA. And fluorescent quantitative PCR assay was used to detect the transcription of fLCF mRNA.
5 Luciferase assays
7.5%CSE treated normal BEAS-2B cells and three kinds of mutant BEAS-2B cells for 24h. And then we measured the luciferase activity.
Results
1 Morphologic change of the BEAS-2B cells treated with CSE
After the stimulated by CSE, the shapes of BEAS-2B cells became irregular. There were more floating cells compared with cells without CSE. We observed a CSE dose dependant increase in cellular damage. And the cell damage became more seriously with the increase of CSE.
2 Changes of green fluorescent protein (EGFP) in three kinds of mutant BEAS-2B cells before and after CSE treated
CSE had an obvious cyto-inhibition. Cells with green fluorescent protein (EGFP) became fewer and round observed under the fluorescence microscope.
3 Results of the MTT colorimetric method
CSE had an obvious cyto-inhibition (F=25.06, P<0.001). With the increase of the concentration of CSE, cyto-inhibition became higher. There was a dose-effect relationship between CSE and the cyto-inhibition. And Inhibitory Concentration 50 of CSE was 7.5%.
4 Transcription of IL-8 mRNA in BEAS-2B cells treated with different concentrations of CSE detected by fluorescent quantitative PCR
BEAS-2B cells were stimulated with 2.5%,5.0%,7.5%and 10.0%CSE for 24h. IL-8 mRNA levels were up-regulated by CSE (2.5%,5.0%and 7.5%), (P<0.001)and had a CSE dose dependent increase. However CSE at a higher concentration (10.0%) made lots of cells dead and did not increase IL-8 mRNA level (P>0.05).
5 Transcription of fLCF mRNA in three kinds of mutant BEAS-2B cells treated with 7.5%CSE detected by fluorescent quantitative PCR
The transcriptions of fLCF mRNA of IL-8mNF-κB-and IL-8mC/EBPβ-BEAS-2B cells were lower than that of IL-8 WT BEAS-2B cells. The transcription of fLCF mRNA of IL-8 WT BEAS-2B cells was 1.32 times than that of IL-8mNF-κB-BEAS-2B cells and 1.54 times than that of IL-8mC/EBPβ-BEAS-2B cells.
6 Luciferase assays
7.5%CSE treated BEAS-2B cells and three kinds of mutant BEAS-2B cells for 24h. We found that the luciferase activities in IL-8mNF-κB-BEAS-2B cells and in IL-8mC/EBPp-BEAS-2B cells were lower than that in IL-8 WT BEAS-2B cells. And the discrepancy had a statistical significance (P< 0.05).
Conclusions
NF-κB regulated the release of Interleukin-8 in BEAS-2B cells treated by cigarette smoke extract. And this study showed for the first time that C/EBPβtranscription factor involved in the precise molecular mechanisms of the release of Interleukin-8 in BEAS-2B cells treated by cigarette smoke extract.
引文
[1]Rennard SI. Cigarette smoke in research [J]. Am J Respir CellMolBiol,2004,31(5):479-480
[2]Church DF, Pryor WA. Free-radical chemistry of cigarette smoke and its toxicological implications [J]. Environ Health Perspect,1985,64:111-126
[3]Cloutier A, T Ear, Blais-Charron, et al. Differential involvement of NF-κB and MAP kinase pathways in the generation of inflammatory cytokines by human neutrophils [J]. Leukocyte Biol,2007,81:567-577
[4]Moretto N, Facchinetti F, Southworth T, et al. alpha, beta-Unsaturated aldehydes contained in cigarette smoke elicit IL-8 release in pulmonary cells through mitogen-activated protein kinases [J]. Am J Physiol Lung Cell Mol Physiol,2009,296(5):L839-48
[5]Shapiro SD, Ingenito EP. The pathogenesis of chronic obstructive pulmonary disease: advances in the past 100 years [J]. Am J Respir Cell Mol Biol,2005,32:367-37
[6]杨丹蕾,徐永健,张珍祥.香烟烟雾提取物对人外周血单个核细胞IL-8 mRNA表达的影响[J].中华微生物学和免疫学杂志,2003,23(7)：557-559
[7]Su Y, Han W, Giraldo C, et al. Effect of cigarette smoke extract on nitric oxide synthase in pulmonary artery endothelial cells [J]. Am J Respir Cell Mol Biol,1998,19:819-25
[8]Dussault A.A., M.Pouliot. Rapid and simple comparison of messenger RNA levels using real-time PCR [J]. Biol Proced Online,2006,8:1-10
[9]Ermann-Lavoie C, Rao CV, Akoum A, et al. Chorionic gonadotrop in down-regulates the expression of the decoy inhibitory interleukin 1 receptor type Ⅱ in human endbmetrial epithelial cells [J]. Endocrinology,2007,148(11):5377
[10]Ear T, McDonald PP. Cytokine generation, promoter activation, and oxidant-independent NF-κB activation in a transfectable human neutrophilic cellular model [J]. BMC Immunol, 2008,9:14
[11]Sasco AJ, Secretan MB, Straif K et al. Tobacco smoking and cancer:a brief review of recent epidemiological evidence [J]. Lung Cancer,2004,45(2):S3-9
[12]Hecht SS. Cigarette smoking and lung cancer and approaches to prevention [J]. Lancet Oncol,2002,3:461-469
[13]Se-Ran Y, Jessica Wright, Mark Bauter, et al. Sirtuin regulates cigarette smoke-induced proinflammatory mediator release via RelA/p65 NFκB in macrophages in vitro and in rat lungs in vivo:implications for chronic inflammation and aging [J]. Am J Physiol Lung Cell Mol Physiol,2007,292:L567-L576
[14]McDonald P.P, A. Bald, M.A. Cassatella, et al. Activation of the NF-κB Pathway by inflammatory stimuli in human neutrophils [J]. Blood,1997,89:3421-3433
[15]Cloutier A, T. Ear, E Borissevitch, et al. Inflammatory cytokine expression is independent of th ec-Jun N-terminal kinase/AP-1 signalin gcascade in human neutrophils [J]. Immunol, 2003,171:3751-3761
[16]Hamilton LM, Davies DE, Wilson SJ, et al. The bronchial epithelium in asthma-much more than a passive barrier [J]. Monaldi Arch Chest Dis,2001,56:48-54
[17]Aruna Kode, Se-Ran Y, Irfan Rahman, et al. Differential effects of cigarette smoke on oxidative stress and proinflammatory cytokine release in primary human airway epithelial cells and in a variety of transformed alveolar epithelial cells [J]. Respiratory Research,2006, 7:132
[18]Pomerantz JL, Baltimore D. Two pathways to NF-kappaB [J]. Mol Cell,2002,10:693-95
[19]Baldin AS Jr. The NF-κB and I-κB proteins:New discoveries and insights [J]. Annu Rev Immunol,1996,14:649
[20]徐祥,梁华平.核因子-KB的结构和功能研究进展[J].细胞与分子遗传学杂志,2002,18：92-95
[21]Rahman I, Gilmour PS, Jimenez LA, et al. Ergothioneine inhibits oxidative stress-and TNF-a-induced NF-κB activation and interleukin-8 release in alveolar epithelial cells [J]. Biochem Biophys Res Commun,2003,302:860-864
[22]Kent L, Smyth L, Clayton C, et al. Cigarette smoke extract induced cytokine and chemokine gene expression changes in COPD macrophages [J]. Cytokine,2008,42(2):205-16
[23]SeRan Yang, Asiya S. Chida, Mark R. Bauter, et al. Cigarette smoke induces proinflammatory cytokine release by activation of NF-B and posttranslational modifications of histone deacetylase in macrophages [J]. Am J Physiol Lung Cell Mol Physiol,2006,291: L46-L57
[24]Ramji D.P., P.Foka. CCAAT/enhancer-binding proteins:structure, function and regulation [J]. J Biol Chem,2002,365:561-575
[25]Twamley-Stein G, E.Kowenz-Leutz, S.Ansieau, et al. Regulation of C/EBP β/NF-M activity by kinase oncogenes [J]. Curr Top Microbiol Immunol,1996,211:129-136
[26]Piwien Pilipuk G, Galigniana MD, Schwartz J. Subnuclear localization of C/EBP beta is regulated by growth hormone and dependent on MAPK [J]. J Biol Chem, 2003,278(37):35668-77
[27]Hongwei G, Shue G,LeLeiko NS, et al. C/EBP y has a stimulatory role on the IL-6 and IL-8 promoters [J]. JBC,2002,12:203-209
[28]Venza 1, Cucinotta M, Visalli M, et al. Pseudomonas aeruginosa induces interleukin-8 (IL-8) gene expression in human conjunctiva through the recruitment of both RelA and CCAAT/enhancer-binding protein beta to the IL-8 promoter [J]. J Biol Chem,2009, 284(7):4191-9
[29]Zhang J, Wu XY, Yu FS. Inflammatory responses of corneal epithelial cells to Pseudomonas aeruginosa infection [J]. Curr Eye Res,2005,30(7):527-34
[30]Wu Q, Lu Z, Verghese MW, et al. Airway epithelial cell tolerance to Pseudomonas aeruginosa [J]. Respir Res,2005,1(6):26
[31]Caristi S, Piraino G,Cucinotta M, et al. Prostaglandin E2 induces interleukin-8 gene transcription by activating C/EBP homologous protein in human T lymphocytes [J]. J Biol Chem,2005,280(15):14433-42
[32]Venza I, Cucinotta M, Caristi S, et al. Transcriptional regulation of IL-8 by Staphylococcus aureus in human conjunctival cells involves activation of AP-1 [J]. Invest Ophthalmol Vis Sci,2007,48(1):270-6
[33]Olive BG, Johnston SL, Baraket M, et al. Increased proinflammatory responses from asthmatic human airway smooth muscle cells in response to rhinovirus infection [J]. Respir Res,2006,7:71
[1]Sasco AJ, Secretan MB, Straif K. Tobacco smoking and cancer:a brief review of recent epidemiological evidence [J]. Lung Cancer,2004,45(2):S3-9
[2]Hecht SS. Cigarette smoking and lung cancer:chemical mechanisms and approaches to prevention [J]. Lancet Oncol,2008,3(8):461-9
[3]李爱兰,黄悦勤,王燕玲,等.我国青少年学生吸烟行为及其影响因素的初步分析[J].中国公共卫生,2001,17(1)：75-77
[4]杨德华.韩松.李敏,等.医学生吸烟行为及其影响因素的研究[J].中国公共卫生,2000,16(5)：437-438
[5]肖锋.减害将推动吸烟新一轮时尚[J].东方烟草报,2004,7(23)
[6]Borgerding M, Klus H. Analysis of complex mixtures-cigarette smoke [J]. Exp Toxicol Pathol,2005,57:43-73
[7]Husgafvel-Pursiainen K. Genotoxicity of environmental tobacco smoke A review [J]. Mutat Res,2004,567(2):427-445
[8]金闻博,戴亚.烟草化学[M].北京：清华大学出版社,1994.103-110
[9]Melkonian G, Cheung L, Marr R, et al. Mainstream and sidestream cigarette smoke inhibit growth and angiogenesis in the day 5 chick chorioallantoic membrane. Toxicol Sci, 2002,68(1):237-48
[10]蔡君兰,赵明月.卷烟主流烟气和侧流烟气中半挥发性成分的分析研究[J].中国烟草学报,2003,9(4)：1-11
[11]李汉超,王淑娴.烟草烟气化学及分析[M].郑州：河南科学技术出版社,1991.59-73
[12]Stedman RL. The chemical composition of tobacco and tobacco smoke [J]. Chem Rev,1968, 68:153
[13]Hoffmann D, Hoffmann I, EL-Bayoumy K. The less harmful cigarette:a controversial issue, a tribute to Ernst L. Wynder [J]. Chem Res Toxicol,2001,14:767-790
[14]Lofroth G, Zebuhr Y. Polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) in mainstream and sidestream cigarette smoke [J]. Bull Environ Contam Toxicol, 1992,48:789-791
[15]闰克玉.卷烟烟气化学[M].郑州：郑州大学出版社,2002.26-3]
[16]王瑞新.烟草化学[M].北京：中国农业出版社,2003.14-18
[17]谢慧玲.不同卷烟主流烟气的细胞毒性及致突变性研究[D].[硕士学位论文].湖南：湖南农业大学,2008
[18]杜咏梅,肖协忠,王允白.烟气焦油与卷烟安全性[J].中国烟草科学,2002,(2)：31-34
[19]Hammond D, Collishaw NE, Callad C. Secret Science Tobacco industry research on smoking behaviour and cigarette toxicity [J]. Lancet,2006,367:781-787
[20]Van Hove CL, Moerloose K, Mase T, et al. Cigarette smoke enhances Th-2 drive airway inflammation and delays inhalational tolerance [J]. Respir Res,2008,9(1):42
[21]Oltmanns U, Chung KF, Walters M, et al. Cigarette smoke induces IL-8, but inhibits eotaxin and RANTES release from airway smooth muscle [J]. Respir Res,2005,6(1):74-83
[22]Luppi F, Aarbiou J, Wetering SV, et al. Effect of cigarette smoke condensate on proliferation and wound closure of bronchial epithelial cells in vitro:role of glutathione [J]. Respir Res, 2005,6(1):140-151
[23]许丽,张珍祥,徐永健.香烟烟雾提取物对气道上皮细胞增殖的影响及其机制[J].中华结核和呼吸杂志,2005,28,2：128-130
[24]Imai K, Mercer BA, Schulman LL, et al. Correlation of lung surface area to apoptosis and proliferation in human emphysema [J]. Eur Respir J,2005,25:250-258
[25]陈剑波,陈平,蔡珊,等.烟雾暴露慢性阻塞性肺疾病大鼠肺内细胞凋亡和增殖的变化[J].中华结核和呼吸杂志,2007,30(9)：709-710
[26]钱革.我国部分地区成人面部皮肤老化相关因素及香烟烟雾提取物与尼古丁致皮肤老化作用和机制研究[D].[博士学位论文].北京：中国协和医科大学,2007
[27]Emster VL, Grady D, Miike R, et al. Facial wrinkling in men and women, by smoking status [J]. AnT J Public Health,1995,85:78-82
[28]Krug M, Wunsche A, Blum A. Addiction to tobacco and the consequences for the skin [J]. Hautarzt,2004,55:301-315
[29]Arredondo J,Hall L L, Ndoye A, et al. Central role of fibroblast alpha3 nicotinic acetylcholine receptor in mediating cutaneous effects of nicotine [J]. Lab Invest,2003, 83:207-225
[30]Northrop-Clewes CA, Thumham DI. Monitoring micronutrients in cigarette smokers [J]. Clin Chim Acta,2007,377:14-38
[31]Lee SD, Lee DS, Chun YG, et al. Cigarette smoke extract induces endothelin-1 via protein kinase C in pulmonary artery endothelial cells [J]. Am J Physiol Lung Cell Mol Physiol, 2001,281(2):L403-L411
[32]Wright JL, Farmer SC, Churg A. A neutrophil elastase inhibitor reduces cigarette smoke induced remodeling of lung vessels [J]. Eur Respir J,2003,22(1):77-81
[33]Santos S, Peinado VI, Ramirez J, et al. Characterization of pulmonary vascular remodeling in smokers and patients with mild COPD [J]. Eur Respir J,2002,19(4):632-638
[34]Levi F, Lucchini F, Negri E. et al. Pancreatic cancer mortality in Europe:the leveling of an epidemic [J]. Pancreas,2003,27:139-142
[35]Wittel UA, Pandey KK, Andria M, et al. Chronic pancreatic inflammation induced by environmental tobacco smoke inhalation in rats [J]. Am J Gastroentero,.2006,101:148-159
[36]Christencen SB. Influence of cigarette smoking on goiter formation [J]. Clin Endocrinol Metab,1984,58:615
[37]Fung H. Postpartum thyroid dysfunction in Mid Glamor-gan [J]. BMJ,1988,296:241-248
[38]Chanoine JP. Smoking during pregnancy:a significant cause of neonatal thyroid enlargement [J]. Br J Obstet Gynaecol,1999,98(1):65-71
[39]杨毅,赵德峰,袁涛,等.环境香烟烟雾对大鼠碘代谢及甲状腺超微结构影响的研究[J].中国公共卫生学报,1997,16(5)：289-290
[40]Bai Q, Xu QQ, Jiang H, et al. Prevalence and risk factors of erectile dysfunction in three cities of China:a communitybased study [J]. Asian J Androl,2004,6(4):343-348
[41]周杏林,邹明杰,吴怡兴,等.吸烟高镉对生精功能的影响[J].中华实验外科杂志,1994,11(4)：203-204
[42]陈晓东,吴玲.维生素C、维生素E拮抗香烟烟雾提取物致小鼠精子畸形效应[J].包头医学院学报,2009,25(2)：136-137
[43]Pasqualotto FF, Sobreiro BP, Hallak J, et al. Cigarette smoking is related to a decrease in semen volume in a population of fertile men [J]. BJU Int,2006,97(2):324-326
[44]Atsushi S, Reiko M, Noriko M, et al. Inhibitory effects of cigarette smoke extract on neural crest migration occur through suppression of R-spondinl expression via aryl hydrocarbon receptor [J]. Naunyn-Schmied Arch Pharmacol,2009,380:569-576
[45]Practice Committee of the American Society for Reproductive Medicine. Smoking and infertility [S]. Fertil Steril,2004,82:S62
[46]Billaud N, Lemarie P. Negative effects of maternal smoking during the course of pregnancy [J]. Arch Pediatr,2001,8:875-877
[47]Hammond D, Collishaw NE, Callad C. Secret Science Tobacco industry research on smoking behaviour and cigarette toxicity [J]. Lancet,2006,367:781-787
[48]Kasai A, Hiramatsu N, Hayakawa K, et al. High levels of dioxin-like potential in cigarette smoke evidenced by in vitro and in vivo biosensing [J]. Cancer Res,2006,66:7143-7148
[49]Mulder I, Hoogenveen RT, Lankisch PG, et al. Smoking cessation would substantially reduce the future incidence of pancreatic cancer in the European Union [J]. Eur J Gastroenterol Hepatol,2002,14:1343-1353
[50]Xu, Talbott EO, Zborowski JV, et al. Cigarette smoking and the risk of adult leukemia: results from the Three Mile Island cohort study [J]. Arch Environ Occup Health,2007, 62(3):131-7
[51]Fernberg T, Odenbro A, Bellocco R, et al. Tobacco use, body mass index, and the risk of leukemia and multiple myeloma:a nationwide cohort study in Sweden [J]. Cancer Res,2007, 67(12):5983-6
[52]Andersson A, Ankerst J, Lindgren A, et al. Hyperhan ocysteinemia and changed plasna thiol redox status in chronic clostructive pulmonary disease [J]. Clin Chem Lab Med,2001, 39:229-233
[53]许建英,庞敏,杜永成,等.香烟烟雾对巨噬细胞的氧化损伤及盐酸氨溴索抗氧化作用的研究[J].中国药物与临床,2006,6(7)：520-522
[54]Jiao ZX, Ao QL, Xiong M. Cigarette smoke extract inhibits the proliferation of alveolar epithelial cells and induces apoptosis [J]. Acta Physiologica Sinica,2006,58(3):244-254
[55]Kirkby KA, Adin CA. Products of heme oxygenase and their potential therapeutic application [J]. Am J Physiol Renal Physiol,2006,290:563-571
[56]Haustein KO, Krause J, Haustein H, et al. Effects of cigarette smoking or nicotine replacement on cardiovascular risk factors and parameters of haemorheology [J]. Intem Med, 2002,252:130-139
[57]孙学敏,张献清,严向阳,等.香烟烟雾提取物对人红细胞膜的损害作用[J].第四军医大学学报,2007,28(18)：1670-1673
[58]Rahman I, Mac Nee W. Oxidative stress and regulation of glutathione in lung inflammation [J]. Eur Respir,.2000,16:534-554
[59]曲仪庆,肖伟,黄绍光,等.香烟提取物和腺病毒E1A基因对肺泡上皮细胞ICAM-1表达的影响[J].中国免疫学杂志,2007,23：886-889
[60]陈芳,陈平,谢丽华,等.香烟烟雾提取物对人内皮细胞细胞间黏附分子-1和白介素-8表达以及与多形核中性粒细胞黏附的影响[J].实用心脑肺血管病杂志,2006,14(12)：942-946
[61]Yin L, Wei G, Deping Zh. Effects of cigarette smoke extract on A549 cells and human lung fibroblasts treated with transforming growth factor-β1 in a coculture system [J]. Clin Exp Med,2009,10:249-251
[62]张晓荣,徐培茹,多力坤.A549核因子-κB和肿瘤坏死因子在香烟烟雾提取物刺激及阿奇霉素干预下的变化[D].[硕士学位论文].新疆：新疆医科大学,2005
[63]Lin JL, Xu YJ, Zhang ZX, et al. Effect of cigarette smoke extract on the role of protein kinase C in the proliferation of passively sensitized human airway smooth muscle cells [J]. Huazhong Univer Sci Technol [Med Sci],2005,25(3):269-273
[64]Kashyap R, Floreania AA, Heires AJ, at al. Protein kinase C-alpha mediates cigarette smoke, extract and complement factor 5a-stimulated interleukin-8 release in human bronchial epithelial cells [J]. Investig Med,2002,50(1):46-53
[65]McKay A, Komai KM, MacLeod K, et al. Interleukin-18 levels in induced sputum are reduced in asthmatic and normal smokers [J]. Clin Exp Allergy,2004,34:904-910
[66]Wong WS, Koh DS. Advances in immunopharmacology of asthma [J]. Biochem Pharmacol, 2000,59:1323-1355
[67]Chakir H, Wang H, Lefebvre DE, et al. T-bet/GATA-3 ratio as a measure of the Th1/Th2 cytokine profile in mixed cell populations:predominant role of GATA-3 [J]. Immunol Methods,2003,278:157-169
[68]Corroyer S, Maitre B, Cazals V, et al. Altered regulation of G1 cyclins in oxidant-induced growth arrest of lung alveolar epithelial cells [J]. J Biol Chem,1996,271(41):25117-25125
[69]Fields WR, Leonard RM, Odom PS, et al. Gene expression in normal human bronchial epithelial (NHBE) cells following in vitro exposure to cigarette smoke condensate [J]. Toxicol Sci,2005,86(1):84-91
[70]Albino AP, Huang X, Jorgensen ED, et al. Induction of DNA double-strand breaks in A549 and normal human pulmonary epithelial cells by cigarette smoke is mediated by free radicals [J]. Int J Oncol,2006,28(6):1491-1505
[71]Huang X, Okafuji M, Traganos F, et al. Assessment of histone H2AX phosphorylation induced by DNA topoisomerase Ⅰ and Ⅱ inhibitors topotecan and mitoxantrone and by the DNA cross-linking agent cisplatin [J]. Cytometry,2004,58(2):99-110
[72]Takai H, Naka K, Suzuki H, et al. Chk2-deficient mice exhibit radioresistance and defective p53-dediated transciption [J]. EMBO J,2002,21(19):5195-5205
[73]Wickenden JA, Clarke MC, Rossi AG, et al. Cigarette smoke prevents apoptosis through inhibition of caspase activation and induces necrosis [J]. Am J Respir Cell Mol Biol,2003, 29(5):562-570
[74]吕兰海,杨华,尤汉虎.卷烟烟雾及其主要有害成分诱发细胞基因突变的研究[J].环境与健康杂志,2004,21(5)：286-288
[75]Vayssier M, Camilli T, Aron Y, et al. Effects of tobacco smoke and benzo[a]pyrene on human endothelial cells and monocyte stress responses [J]. Am J Physiol Heart Circ Physiol, 2001,280(3):1293-1298
[76]孙咏梅,戴树桂,袭著革.香烟烟雾成分分析及其对DNA生物氧化能力研究[J].环境与健康杂志,2001,18(4)：203-207
[77]Hei Tk. Radom-induced neoplastic transformation of human bronchial epithelial cells [J]. Radi Oncol Inv,1996,3:398-404
[78]Suzuki K, Hei TK. Mutation induction in gamma-irradiated primary human bronchial epithelial cells and molecular analysis of the HPRT-mutants [J]. Mutat Res,1996,349:33-41
[79]李继红.香烟提取物对人体NK细胞和LAK细胞活性影响[J].中国公共卫生,2005,21(2)：179-18
[2]Church DF, Pryor WA. Free-radical chemistry of cigarette smoke and its toxicological implications [J]. Environ Health Perspect,1985,64:111-126
[3]Cloutier A, T Ear, Blais-Charron, et al. Differential involvement of NF-κB and MAP kinase pathways in the generation of inflammatory cytokines by human neutrophils [J]. Leukocyte Biol,2007,81:567-577
[4]Moretto N, Facchinetti F, Southworth T, et al. alpha, beta-Unsaturated aldehydes contained in cigarette smoke elicit IL-8 release in pulmonary cells through mitogen-activated protein kinases [J]. Am J Physiol Lung Cell Mol Physiol,2009,296(5):L839-48
[5]Shapiro SD, Ingenito EP. The pathogenesis of chronic obstructive pulmonary disease: advances in the past 100 years [J]. Am J Respir Cell Mol Biol,2005,32:367-37
[6]杨丹蕾,徐永健,张珍祥.香烟烟雾提取物对人外周血单个核细胞IL-8 mRNA表达的影响[J].中华微生物学和免疫学杂志,2003,23(7)：557-559
[7]Su Y, Han W, Giraldo C, et al. Effect of cigarette smoke extract on nitric oxide synthase in pulmonary artery endothelial cells [J]. Am J Respir Cell Mol Biol,1998,19:819-25
[8]Dussault A.A., M.Pouliot. Rapid and simple comparison of messenger RNA levels using real-time PCR [J]. Biol Proced Online,2006,8:1-10
[9]Ermann-Lavoie C, Rao CV, Akoum A, et al. Chorionic gonadotrop in down-regulates the expression of the decoy inhibitory interleukin 1 receptor type Ⅱ in human endbmetrial epithelial cells [J]. Endocrinology,2007,148(11):5377
[10]Ear T, McDonald PP. Cytokine generation, promoter activation, and oxidant-independent NF-κB activation in a transfectable human neutrophilic cellular model [J]. BMC Immunol, 2008,9:14
[11]Sasco AJ, Secretan MB, Straif K et al. Tobacco smoking and cancer:a brief review of recent epidemiological evidence [J]. Lung Cancer,2004,45(2):S3-9
[12]Hecht SS. Cigarette smoking and lung cancer and approaches to prevention [J]. Lancet Oncol,2002,3:461-469
[13]Se-Ran Y, Jessica Wright, Mark Bauter, et al. Sirtuin regulates cigarette smoke-induced proinflammatory mediator release via RelA/p65 NFκB in macrophages in vitro and in rat lungs in vivo:implications for chronic inflammation and aging [J]. Am J Physiol Lung Cell Mol Physiol,2007,292:L567-L576
[14]McDonald P.P, A. Bald, M.A. Cassatella, et al. Activation of the NF-κB Pathway by inflammatory stimuli in human neutrophils [J]. Blood,1997,89:3421-3433
[15]Cloutier A, T. Ear, E Borissevitch, et al. Inflammatory cytokine expression is independent of th ec-Jun N-terminal kinase/AP-1 signalin gcascade in human neutrophils [J]. Immunol, 2003,171:3751-3761
[16]Hamilton LM, Davies DE, Wilson SJ, et al. The bronchial epithelium in asthma-much more than a passive barrier [J]. Monaldi Arch Chest Dis,2001,56:48-54
[17]Aruna Kode, Se-Ran Y, Irfan Rahman, et al. Differential effects of cigarette smoke on oxidative stress and proinflammatory cytokine release in primary human airway epithelial cells and in a variety of transformed alveolar epithelial cells [J]. Respiratory Research,2006, 7:132
[18]Pomerantz JL, Baltimore D. Two pathways to NF-kappaB [J]. Mol Cell,2002,10:693-95
[19]Baldin AS Jr. The NF-κB and I-κB proteins:New discoveries and insights [J]. Annu Rev Immunol,1996,14:649
[20]徐祥,梁华平.核因子-KB的结构和功能研究进展[J].细胞与分子遗传学杂志,2002,18：92-95
[21]Rahman I, Gilmour PS, Jimenez LA, et al. Ergothioneine inhibits oxidative stress-and TNF-a-induced NF-κB activation and interleukin-8 release in alveolar epithelial cells [J]. Biochem Biophys Res Commun,2003,302:860-864
[22]Kent L, Smyth L, Clayton C, et al. Cigarette smoke extract induced cytokine and chemokine gene expression changes in COPD macrophages [J]. Cytokine,2008,42(2):205-16
[23]SeRan Yang, Asiya S. Chida, Mark R. Bauter, et al. Cigarette smoke induces proinflammatory cytokine release by activation of NF-B and posttranslational modifications of histone deacetylase in macrophages [J]. Am J Physiol Lung Cell Mol Physiol,2006,291: L46-L57
[24]Ramji D.P., P.Foka. CCAAT/enhancer-binding proteins:structure, function and regulation [J]. J Biol Chem,2002,365:561-575
[25]Twamley-Stein G, E.Kowenz-Leutz, S.Ansieau, et al. Regulation of C/EBP β/NF-M activity by kinase oncogenes [J]. Curr Top Microbiol Immunol,1996,211:129-136
[26]Piwien Pilipuk G, Galigniana MD, Schwartz J. Subnuclear localization of C/EBP beta is regulated by growth hormone and dependent on MAPK [J]. J Biol Chem, 2003,278(37):35668-77
[27]Hongwei G, Shue G,LeLeiko NS, et al. C/EBP y has a stimulatory role on the IL-6 and IL-8 promoters [J]. JBC,2002,12:203-209
[28]Venza 1, Cucinotta M, Visalli M, et al. Pseudomonas aeruginosa induces interleukin-8 (IL-8) gene expression in human conjunctiva through the recruitment of both RelA and CCAAT/enhancer-binding protein beta to the IL-8 promoter [J]. J Biol Chem,2009, 284(7):4191-9
[29]Zhang J, Wu XY, Yu FS. Inflammatory responses of corneal epithelial cells to Pseudomonas aeruginosa infection [J]. Curr Eye Res,2005,30(7):527-34
[30]Wu Q, Lu Z, Verghese MW, et al. Airway epithelial cell tolerance to Pseudomonas aeruginosa [J]. Respir Res,2005,1(6):26
[31]Caristi S, Piraino G,Cucinotta M, et al. Prostaglandin E2 induces interleukin-8 gene transcription by activating C/EBP homologous protein in human T lymphocytes [J]. J Biol Chem,2005,280(15):14433-42
[32]Venza I, Cucinotta M, Caristi S, et al. Transcriptional regulation of IL-8 by Staphylococcus aureus in human conjunctival cells involves activation of AP-1 [J]. Invest Ophthalmol Vis Sci,2007,48(1):270-6
[33]Olive BG, Johnston SL, Baraket M, et al. Increased proinflammatory responses from asthmatic human airway smooth muscle cells in response to rhinovirus infection [J]. Respir Res,2006,7:71
[1]Sasco AJ, Secretan MB, Straif K. Tobacco smoking and cancer:a brief review of recent epidemiological evidence [J]. Lung Cancer,2004,45(2):S3-9
[2]Hecht SS. Cigarette smoking and lung cancer:chemical mechanisms and approaches to prevention [J]. Lancet Oncol,2008,3(8):461-9
[3]李爱兰,黄悦勤,王燕玲,等.我国青少年学生吸烟行为及其影响因素的初步分析[J].中国公共卫生,2001,17(1)：75-77
[4]杨德华.韩松.李敏,等.医学生吸烟行为及其影响因素的研究[J].中国公共卫生,2000,16(5)：437-438
[5]肖锋.减害将推动吸烟新一轮时尚[J].东方烟草报,2004,7(23)
[6]Borgerding M, Klus H. Analysis of complex mixtures-cigarette smoke [J]. Exp Toxicol Pathol,2005,57:43-73
[7]Husgafvel-Pursiainen K. Genotoxicity of environmental tobacco smoke A review [J]. Mutat Res,2004,567(2):427-445
[8]金闻博,戴亚.烟草化学[M].北京：清华大学出版社,1994.103-110
[9]Melkonian G, Cheung L, Marr R, et al. Mainstream and sidestream cigarette smoke inhibit growth and angiogenesis in the day 5 chick chorioallantoic membrane. Toxicol Sci, 2002,68(1):237-48
[10]蔡君兰,赵明月.卷烟主流烟气和侧流烟气中半挥发性成分的分析研究[J].中国烟草学报,2003,9(4)：1-11
[11]李汉超,王淑娴.烟草烟气化学及分析[M].郑州：河南科学技术出版社,1991.59-73
[12]Stedman RL. The chemical composition of tobacco and tobacco smoke [J]. Chem Rev,1968, 68:153
[13]Hoffmann D, Hoffmann I, EL-Bayoumy K. The less harmful cigarette:a controversial issue, a tribute to Ernst L. Wynder [J]. Chem Res Toxicol,2001,14:767-790
[14]Lofroth G, Zebuhr Y. Polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) in mainstream and sidestream cigarette smoke [J]. Bull Environ Contam Toxicol, 1992,48:789-791
[15]闰克玉.卷烟烟气化学[M].郑州：郑州大学出版社,2002.26-3]
[16]王瑞新.烟草化学[M].北京：中国农业出版社,2003.14-18
[17]谢慧玲.不同卷烟主流烟气的细胞毒性及致突变性研究[D].[硕士学位论文].湖南：湖南农业大学,2008
[18]杜咏梅,肖协忠,王允白.烟气焦油与卷烟安全性[J].中国烟草科学,2002,(2)：31-34
[19]Hammond D, Collishaw NE, Callad C. Secret Science Tobacco industry research on smoking behaviour and cigarette toxicity [J]. Lancet,2006,367:781-787
[20]Van Hove CL, Moerloose K, Mase T, et al. Cigarette smoke enhances Th-2 drive airway inflammation and delays inhalational tolerance [J]. Respir Res,2008,9(1):42
[21]Oltmanns U, Chung KF, Walters M, et al. Cigarette smoke induces IL-8, but inhibits eotaxin and RANTES release from airway smooth muscle [J]. Respir Res,2005,6(1):74-83
[22]Luppi F, Aarbiou J, Wetering SV, et al. Effect of cigarette smoke condensate on proliferation and wound closure of bronchial epithelial cells in vitro:role of glutathione [J]. Respir Res, 2005,6(1):140-151
[23]许丽,张珍祥,徐永健.香烟烟雾提取物对气道上皮细胞增殖的影响及其机制[J].中华结核和呼吸杂志,2005,28,2：128-130
[24]Imai K, Mercer BA, Schulman LL, et al. Correlation of lung surface area to apoptosis and proliferation in human emphysema [J]. Eur Respir J,2005,25:250-258
[25]陈剑波,陈平,蔡珊,等.烟雾暴露慢性阻塞性肺疾病大鼠肺内细胞凋亡和增殖的变化[J].中华结核和呼吸杂志,2007,30(9)：709-710
[26]钱革.我国部分地区成人面部皮肤老化相关因素及香烟烟雾提取物与尼古丁致皮肤老化作用和机制研究[D].[博士学位论文].北京：中国协和医科大学,2007
[27]Emster VL, Grady D, Miike R, et al. Facial wrinkling in men and women, by smoking status [J]. AnT J Public Health,1995,85:78-82
[28]Krug M, Wunsche A, Blum A. Addiction to tobacco and the consequences for the skin [J]. Hautarzt,2004,55:301-315
[29]Arredondo J,Hall L L, Ndoye A, et al. Central role of fibroblast alpha3 nicotinic acetylcholine receptor in mediating cutaneous effects of nicotine [J]. Lab Invest,2003, 83:207-225
[30]Northrop-Clewes CA, Thumham DI. Monitoring micronutrients in cigarette smokers [J]. Clin Chim Acta,2007,377:14-38
[31]Lee SD, Lee DS, Chun YG, et al. Cigarette smoke extract induces endothelin-1 via protein kinase C in pulmonary artery endothelial cells [J]. Am J Physiol Lung Cell Mol Physiol, 2001,281(2):L403-L411
[32]Wright JL, Farmer SC, Churg A. A neutrophil elastase inhibitor reduces cigarette smoke induced remodeling of lung vessels [J]. Eur Respir J,2003,22(1):77-81
[33]Santos S, Peinado VI, Ramirez J, et al. Characterization of pulmonary vascular remodeling in smokers and patients with mild COPD [J]. Eur Respir J,2002,19(4):632-638
[34]Levi F, Lucchini F, Negri E. et al. Pancreatic cancer mortality in Europe:the leveling of an epidemic [J]. Pancreas,2003,27:139-142
[35]Wittel UA, Pandey KK, Andria M, et al. Chronic pancreatic inflammation induced by environmental tobacco smoke inhalation in rats [J]. Am J Gastroentero,.2006,101:148-159
[36]Christencen SB. Influence of cigarette smoking on goiter formation [J]. Clin Endocrinol Metab,1984,58:615
[37]Fung H. Postpartum thyroid dysfunction in Mid Glamor-gan [J]. BMJ,1988,296:241-248
[38]Chanoine JP. Smoking during pregnancy:a significant cause of neonatal thyroid enlargement [J]. Br J Obstet Gynaecol,1999,98(1):65-71
[39]杨毅,赵德峰,袁涛,等.环境香烟烟雾对大鼠碘代谢及甲状腺超微结构影响的研究[J].中国公共卫生学报,1997,16(5)：289-290
[40]Bai Q, Xu QQ, Jiang H, et al. Prevalence and risk factors of erectile dysfunction in three cities of China:a communitybased study [J]. Asian J Androl,2004,6(4):343-348
[41]周杏林,邹明杰,吴怡兴,等.吸烟高镉对生精功能的影响[J].中华实验外科杂志,1994,11(4)：203-204
[42]陈晓东,吴玲.维生素C、维生素E拮抗香烟烟雾提取物致小鼠精子畸形效应[J].包头医学院学报,2009,25(2)：136-137
[43]Pasqualotto FF, Sobreiro BP, Hallak J, et al. Cigarette smoking is related to a decrease in semen volume in a population of fertile men [J]. BJU Int,2006,97(2):324-326
[44]Atsushi S, Reiko M, Noriko M, et al. Inhibitory effects of cigarette smoke extract on neural crest migration occur through suppression of R-spondinl expression via aryl hydrocarbon receptor [J]. Naunyn-Schmied Arch Pharmacol,2009,380:569-576
[45]Practice Committee of the American Society for Reproductive Medicine. Smoking and infertility [S]. Fertil Steril,2004,82:S62
[46]Billaud N, Lemarie P. Negative effects of maternal smoking during the course of pregnancy [J]. Arch Pediatr,2001,8:875-877
[47]Hammond D, Collishaw NE, Callad C. Secret Science Tobacco industry research on smoking behaviour and cigarette toxicity [J]. Lancet,2006,367:781-787
[48]Kasai A, Hiramatsu N, Hayakawa K, et al. High levels of dioxin-like potential in cigarette smoke evidenced by in vitro and in vivo biosensing [J]. Cancer Res,2006,66:7143-7148
[49]Mulder I, Hoogenveen RT, Lankisch PG, et al. Smoking cessation would substantially reduce the future incidence of pancreatic cancer in the European Union [J]. Eur J Gastroenterol Hepatol,2002,14:1343-1353
[50]Xu, Talbott EO, Zborowski JV, et al. Cigarette smoking and the risk of adult leukemia: results from the Three Mile Island cohort study [J]. Arch Environ Occup Health,2007, 62(3):131-7
[51]Fernberg T, Odenbro A, Bellocco R, et al. Tobacco use, body mass index, and the risk of leukemia and multiple myeloma:a nationwide cohort study in Sweden [J]. Cancer Res,2007, 67(12):5983-6
[52]Andersson A, Ankerst J, Lindgren A, et al. Hyperhan ocysteinemia and changed plasna thiol redox status in chronic clostructive pulmonary disease [J]. Clin Chem Lab Med,2001, 39:229-233
[53]许建英,庞敏,杜永成,等.香烟烟雾对巨噬细胞的氧化损伤及盐酸氨溴索抗氧化作用的研究[J].中国药物与临床,2006,6(7)：520-522
[54]Jiao ZX, Ao QL, Xiong M. Cigarette smoke extract inhibits the proliferation of alveolar epithelial cells and induces apoptosis [J]. Acta Physiologica Sinica,2006,58(3):244-254
[55]Kirkby KA, Adin CA. Products of heme oxygenase and their potential therapeutic application [J]. Am J Physiol Renal Physiol,2006,290:563-571
[56]Haustein KO, Krause J, Haustein H, et al. Effects of cigarette smoking or nicotine replacement on cardiovascular risk factors and parameters of haemorheology [J]. Intem Med, 2002,252:130-139
[57]孙学敏,张献清,严向阳,等.香烟烟雾提取物对人红细胞膜的损害作用[J].第四军医大学学报,2007,28(18)：1670-1673
[58]Rahman I, Mac Nee W. Oxidative stress and regulation of glutathione in lung inflammation [J]. Eur Respir,.2000,16:534-554
[59]曲仪庆,肖伟,黄绍光,等.香烟提取物和腺病毒E1A基因对肺泡上皮细胞ICAM-1表达的影响[J].中国免疫学杂志,2007,23：886-889
[60]陈芳,陈平,谢丽华,等.香烟烟雾提取物对人内皮细胞细胞间黏附分子-1和白介素-8表达以及与多形核中性粒细胞黏附的影响[J].实用心脑肺血管病杂志,2006,14(12)：942-946
[61]Yin L, Wei G, Deping Zh. Effects of cigarette smoke extract on A549 cells and human lung fibroblasts treated with transforming growth factor-β1 in a coculture system [J]. Clin Exp Med,2009,10:249-251
[62]张晓荣,徐培茹,多力坤.A549核因子-κB和肿瘤坏死因子在香烟烟雾提取物刺激及阿奇霉素干预下的变化[D].[硕士学位论文].新疆：新疆医科大学,2005
[63]Lin JL, Xu YJ, Zhang ZX, et al. Effect of cigarette smoke extract on the role of protein kinase C in the proliferation of passively sensitized human airway smooth muscle cells [J]. Huazhong Univer Sci Technol [Med Sci],2005,25(3):269-273
[64]Kashyap R, Floreania AA, Heires AJ, at al. Protein kinase C-alpha mediates cigarette smoke, extract and complement factor 5a-stimulated interleukin-8 release in human bronchial epithelial cells [J]. Investig Med,2002,50(1):46-53
[65]McKay A, Komai KM, MacLeod K, et al. Interleukin-18 levels in induced sputum are reduced in asthmatic and normal smokers [J]. Clin Exp Allergy,2004,34:904-910
[66]Wong WS, Koh DS. Advances in immunopharmacology of asthma [J]. Biochem Pharmacol, 2000,59:1323-1355
[67]Chakir H, Wang H, Lefebvre DE, et al. T-bet/GATA-3 ratio as a measure of the Th1/Th2 cytokine profile in mixed cell populations:predominant role of GATA-3 [J]. Immunol Methods,2003,278:157-169
[68]Corroyer S, Maitre B, Cazals V, et al. Altered regulation of G1 cyclins in oxidant-induced growth arrest of lung alveolar epithelial cells [J]. J Biol Chem,1996,271(41):25117-25125
[69]Fields WR, Leonard RM, Odom PS, et al. Gene expression in normal human bronchial epithelial (NHBE) cells following in vitro exposure to cigarette smoke condensate [J]. Toxicol Sci,2005,86(1):84-91
[70]Albino AP, Huang X, Jorgensen ED, et al. Induction of DNA double-strand breaks in A549 and normal human pulmonary epithelial cells by cigarette smoke is mediated by free radicals [J]. Int J Oncol,2006,28(6):1491-1505
[71]Huang X, Okafuji M, Traganos F, et al. Assessment of histone H2AX phosphorylation induced by DNA topoisomerase Ⅰ and Ⅱ inhibitors topotecan and mitoxantrone and by the DNA cross-linking agent cisplatin [J]. Cytometry,2004,58(2):99-110
[72]Takai H, Naka K, Suzuki H, et al. Chk2-deficient mice exhibit radioresistance and defective p53-dediated transciption [J]. EMBO J,2002,21(19):5195-5205
[73]Wickenden JA, Clarke MC, Rossi AG, et al. Cigarette smoke prevents apoptosis through inhibition of caspase activation and induces necrosis [J]. Am J Respir Cell Mol Biol,2003, 29(5):562-570
[74]吕兰海,杨华,尤汉虎.卷烟烟雾及其主要有害成分诱发细胞基因突变的研究[J].环境与健康杂志,2004,21(5)：286-288
[75]Vayssier M, Camilli T, Aron Y, et al. Effects of tobacco smoke and benzo[a]pyrene on human endothelial cells and monocyte stress responses [J]. Am J Physiol Heart Circ Physiol, 2001,280(3):1293-1298
[76]孙咏梅,戴树桂,袭著革.香烟烟雾成分分析及其对DNA生物氧化能力研究[J].环境与健康杂志,2001,18(4)：203-207
[77]Hei Tk. Radom-induced neoplastic transformation of human bronchial epithelial cells [J]. Radi Oncol Inv,1996,3:398-404
[78]Suzuki K, Hei TK. Mutation induction in gamma-irradiated primary human bronchial epithelial cells and molecular analysis of the HPRT-mutants [J]. Mutat Res,1996,349:33-41
[79]李继红.香烟提取物对人体NK细胞和LAK细胞活性影响[J].中国公共卫生,2005,21(2)：179-18