摘要
采用C57BL6小鼠作为模型,SO2（7mg/m3）动式吸入染毒28d,每天染毒6h;SO2熏气开始后的第1~5d,腹腔注射Ba P（40mg/kg b.w.）,一天一次.吸入染毒结束后,采用分光光度计法检测小鼠肺线粒体膜电位（MMP）以及细胞色素C氧化酶（COX）的活性;采用荧光定量PCR技术,检测肺细胞中细胞色素C氧化酶亚基CO4以及核转录因子PGC1-α、NRF1、mtTFA的mRNA水平;并采用Western blot技术检测上述三种线粒体调控基因的蛋白表达.结果发现,在SO2和BaP复合暴露后,小鼠肺线粒体膜电位下降,氧化磷酸化复合体COX活性和亚基CO4的mRNA表达水平显著降低,调控基因NRF1和mtTFA的mRNA和蛋白水平也显著下调.提示小鼠在SO2和BaP复合暴露后,可能通过降低肺中NRF1表达,影响其对mtTFA的调控,进一步抑制相关基因组的转录和翻译,最终可能导致小鼠肺线粒体的氧化磷酸化功能受损,线粒体膜电位下降进而引发肺部疾病.
C57BL6 male mice were exposed to SO2（7mg/m3） for 28 days, 6hours per day. In the first 5days of SO2 inhalation, mice were injected intraperitoneally with Ba P（40mg/kg b. w.）, which was dissolved in oil. The mitochondrial membrane potential（MMP） and activity of cytochrome C oxidase（COX） were detected by spectrophotometer. The m RNA levels of one subunit of COX（CO4） and three mitochondrial transcript factors（PGC1-α, NRF1, mt TFA） were analyzed by real-time RT-PCR after exposure. And the protein levels of the above three mitochondrial transcript factors were detected by Western blot. The results demonstrated that the co-exposure of SO2 and Ba P statistically reduced the MMP and activity of COX. The m RNA expression of CO4 was decreased after co-exposure of SO2 and Ba P, combined with the depressions of NRF1 and mt TFA m RNA and protein levels. It is indicated that conjunction of SO2 and Ba P regulated the expression of NRF1, which then inhibited the transcription and translation of related genomes and resulted in mitochondria damage in mouse lung.
出处
《中国环境科学》
EI
CAS
CSCD
北大核心
2015年第11期3496-3501,共6页
China Environmental Science
基金
国家自然科学基金(21007036)
山西省自然科学基金(2012011036-6)
山西省高校优秀青年学术带头人支持计划(20120303)
