摘要
目的寻找与大气污染致病相关的相关基因,为阐明大气污染致病的生物学机制提供科学依据。方法采用mRNA斑点杂交鉴定技术,克隆经采暖季≥75μg/m3 PM2.5与非采暖季<75μg/m3 PM2.5染毒的WI-38人胚肺细胞,分析其间的基因表达差异。放免法测定炎性因子TNF-α、IL-2、IL-6和IL-8。结果与对照组比较,PM2.5>100μg/mL染毒24h后,WI-38人胚肺细胞细胞因子TNF-α、IL-6和IL-8明显升高,IL-2明显减低(P<0.05)。不同浓度PM2.5处理的WI-38人胚肺细胞间差异表达的基因片段,可见48份基因样本在350bp处出现清晰条带;该48份基因样本经斑点杂交后,Tester cDNA杂交的48个斑点中,41份可见黑褐色斑点,而同样样本与Driver cDNA杂交的该48份样本均未见明显显色。结论 PM2.5能诱导WI-38人胚肺细胞产生炎性损伤,≥75μg/m3 PM2.5染毒的WI-38人胚肺细胞存在明显的基因损伤。
Objective To investigate the pathogen-related genes of atmospheric polluting disease so as to clarify the biology mechanism and provide the scientific basis.Methods By using the technique of dot blot hybridization,we analyzed genes＇differential expression with cloning by exposure to ≥75μg/m3 PM2.5 in heating season and 75μg/m3 PM2.5 in un-heating season in WI-38 human embryo lung cells.The levels of cytokines TNF-α,IL-2,IL-6 and IL-8 were determined by radio immunity assay.Results After 24 h of treatment,compared with control group,more than100μg/mL PM2.5significantly increased TNF-α,IL-6 and IL-8levels,and decreased IL-2 in WI-38 human embryo lung cells（P〈0.05）.The clear stripe was found in 350 bp in 48 gene samples with segment with differential expression of genes exposed to different concentrations of PM2.5 in WI-38 human embryo lung cells.Through the dot blot hybridization,black brown spots were found in41 samples in Tester cDNA hybridization,and no similar spots were found in all of the same samples in Driver cDNA hybridization.Conclusion PM2.5 exposure may induce the inflammatory damage of WI-38 human embryo lung cells.Obvious genetic damage was observed in those cells exposed to ≥75μg/m3 PM2.5 in heating season.
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2015年第6期836-839,共4页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
2013年国家级大学生创新训练项目(No.201310698066)~~
