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香烟烟雾暴露对小鼠下呼吸道菌群的影响
作者:陈玲 李康杰 黄尧 陈子龙 张锐 李革 
单位:400016 重庆 重庆医科大学实验教学管理中心(陈玲、李革) 公共卫生与管理学院(李康杰、张锐) 第一临床学院(黄尧、陈子龙) 
关键词:吸烟 烟草 呼吸道疾病 微生物群 下呼吸道 
分类号:R563.9
出版年,卷(期):页码:2019,44(9):739-744
摘要:

 [摘要]  目的  探究香烟烟雾暴露对小鼠下呼吸道菌群的影响。方法  将40只清洁级雄性昆明系小鼠按体重随机区组法分为染毒组、对照组两个体内实验组,每组20只;另外设1个体外实验组(n=20)。染毒组小鼠给予每天香烟烟雾染毒2 h,每次14支香烟,对照组和体外实验组小鼠不做处理,90 d后断颈处死小鼠。采用Mueller-Hinton Broth(MHB)逐级稀释法配制不同浓度梯度的香烟烟雾溶液(CSS),加入体外实验小鼠的肺部灌洗液,观察小鼠下呼吸道菌群的生长情况。采用HE染色观察染毒组与对照组小鼠肺组织病理学变化;采用16S rRNA测序法检测小鼠下呼吸道菌群结构。结果  CSS对体外小鼠下呼吸道菌群的最低抑制浓度约为14.43 μg/ml。当培养基中CSS浓度14.43 μg/ml 时,抑菌率均为0,细菌生长旺盛;当培养基中CSS浓度14.43 μg/ml时,抑菌率均高达90%以上。HE染色示染毒组小鼠肺组织发生明显的炎性改变。染毒组与对照组小鼠下呼吸道菌群相同的操作分类单元(OTUs)856种,染毒组特有OTUs 601种,对照组特有OTUs 422种。从细菌门水平上分析,染毒组小鼠下呼吸道中的优势菌门为变形菌和硬壁菌,其相对丰度分别为61.64%±8.21%24.00%±8.23%,而对照组分别为62.96%±4.32%23.51%±3.39%,两组比较差异无统计学意义(P0.05)。从细菌属水平上分析,染毒组与对照组的优势菌属均为盐单胞菌,其相对丰度分别为20.01%±7.15%21.98%±4.00%,两组比较差异无统计学意义(P0.05)结论  香烟烟雾暴露可能通过改变下呼吸道菌群结构引起宿主下呼吸道免疫炎症反应,从而导致或者促进呼吸系统疾病的发生。

 [Abstracts]  Objective  To investigate the effect of tobacco smoke exposure on the lower respiratory tract flora in mice. Methods  Forty Kun-Ming male mice were randomly divided into smoking group and non-smoking group (20 each) according to their weight: In addition, in vitro group was set (n=20). Mice in smoking group were exposed to tobacco smoke 2 h/d for 90 days, and in non-smoking group and in vitro group were not treated. All mice were sacrificed 90 days later. Different concentrations of gradient cigarette smoke solutions (CSS) were configured by Mueller-Hinton Broth (MHB) step-by-step dilution method. The growth of lower respiratory tract flora was observed in mice of in vitro group by adding lung lavage. The pathological changes of lung tissues of mice in smoking group and non-smoking group were observed by hematoxylin-eosin staining. The development of lower respiratory tract flora in mice was detected by 16S rRNA sequencing. Results  The minimum inhibitory concentration of CSS on the lower respiratory tract flora in mice of in vitro group was about 14.43 μg/ml. When CSS concentration was <14.43 μg/ ml, the growth of bacteria was strong. When CSS concentration was 14.43 μg/ml, the antibacterial rate was over 90%. HE staining showed obvious inflammatory changes occurred in lung tissue of mice in smoking group. There were 856 species with same OTUs in lower respiratory tract flora of both smoking group and control group, 601 species with special OTUs in smoking group and 422 species with special OTUs in non-smoking group. On the phylum level, the dominant bacteria in smoking group were Proteobacteria and Firmicutes, the relative abundance in smoking group were 61.64%±8.21% and 24.00%±8.23%, respectively, and in non-smoking group were 62.96%±4.32% and 23.51%±3.39%, respectively. There were no significant differences between the smoking group and control group (P0.05). On the genus level, the dominant bacteria in both smoking group and non-smoking group was Halomonas, with the relative abundances of 20.01%±7.15% and 21.98%±4.00%, respectively. There was no significant difference between the smoking group and control group (P0.05). Conclusion  Cigarette smoke exposure may cause inflammation in the lower respiratory tract of host by altering the structure of the lower respiratory tract flora, leading to or promoting the occurrence of respiratory diseases.

基金项目:
重庆医科大学实验教学管理中心“MTS”项目(LTMCMTS201901)
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