CAJ | 학술논문

目的利用基因芯片技术检测大鼠孕期苯并[a]芘暴露后仔鼠肝细胞基因表达谱的变化,探讨对仔鼠肝细胞毒作用机制中具有核心调控作用的基因和信号通路.方法选取体重220～250 g SPF级雌、雄性SD大鼠各32只,以随机数字法1∶1比例合笼,将受孕雌鼠分为低、中、高3个染毒组和1个溶剂对照组,每组8只.在孕期第3 ～17天连续灌胃,苯并[a]芘染毒浓度分别为0.75、1.50、3.00 mg/kg,构建大鼠孕期苯并[a]芘的暴露模型.孕鼠正常分娩后取新生仔鼠肝脏,使用RatRef-12芯片检测各组肝细胞全基因组表达情况,并进行重复实验以评价芯片检测的稳定性.结果每组采用单纯随机法选取来自3只孕鼠的仔鼠肝组织,经过基因芯片检测发现新生仔鼠肝脏有1232个基因发生变化；并发现相关的基因出现了3种较典型的表达趋势(P＜0.05),分别为随着苯并[a]芘染毒剂量的增加,基因出现表达上调、下调和波动性变化.所涉及的差异具有统计学意义的信号通路26条(P＜0.05),主要集中于生长发育信号通路、毒物代谢信号通路和炎症信号通路.在差异基因构建的整体网络中,以CYP2C13、GSTO1基因为核心的代谢通路,以Rela基因为核心的凋亡通路及以MAPK8和Plcg1基因为核心的生长发育通路是孕期苯并[a]芘染毒导致仔鼠肝细胞毒性的关键核心通路.结论孕期苯并[a]芘暴露对子代肝细胞基因表达产生了一定的影响,发现了一些具有核心调控作用的基因及相关信号通路,为探讨苯并[a]芘暴露对子代肝细胞的影响及其毒作用机制提供了一定的依据.
목적 이용기인심편기술검측대서잉기분병[a]비폭로후자서간세포기인표체보적변화,탐토대자서간세포독작용궤제중구유핵심조공작용적기인화신호통로.방법 선취체중220～250 g SPF급자、웅성SD대서각32지,이수궤수자법1∶1비례합롱,장수잉자서분위저、중、고3개염독조화1개용제대조조,매조8지.재잉기제3 ～17천련속관위,분병[a]비염독농도분별위0.75、1.50、3.00 mg/kg,구건대서잉기분병[a]비적폭로모형.잉서정상분면후취신생자서간장,사용RatRef-12심편검측각조간세포전기인조표체정황,병진행중복실험이평개심편검측적은정성.결과 매조채용단순수궤법선취래자3지잉서적자서간조직,경과기인심편검측발현신생자서간장유1232개기인발생변화；병발현상관적기인출현료3충교전형적표체추세(P＜0.05),분별위수착분병[a]비염독제량적증가,기인출현표체상조、하조화파동성변화.소섭급적차이구유통계학의의적신호통로26조(P＜0.05),주요집중우생장발육신호통로、독물대사신호통로화염증신호통로.재차이기인구건적정체망락중,이CYP2C13、GSTO1기인위핵심적대사통로,이Rela기인위핵심적조망통로급이MAPK8화Plcg1기인위핵심적생장발육통로시잉기분병[a]비염독도치자서간세포독성적관건핵심통로.결론 잉기분병[a]비폭로대자대간세포기인표체산생료일정적영향,발현료일사구유핵심조공작용적기인급상관신호통로,위탐토분병[a]비폭로대자대간세포적영향급기독작용궤제제공료일정적의거.
Objective To study the gene expression patterns in livers of infant rats after Benzo[a]pyrene (BaP) exposure during pregnancy and explore the important gene and signaling pathways in the toxic mechanism of BaP.Methods Thirty-two pregnant SD rats were randomly divided into four groups:vehicle control (corn oil) and treatment groups (0.75,1.50 and 3.00 mg/kg BaP in corn oil).BaP solutions were given by gastric infusion from the 3rd to the 17th day of pregnancy.After delivery the offspring's liver were taken to detect the gene expression by RatRef-12 gene chip.The stability of gene chip was tested by repeated experiments.Results After prenatal BaP exposure 1232 genes with different expression variations in hepatocytes of offsprings were identified.Three expression patterns of genes related to the dose of prenatal BaP exposure were identified with significant difference (P ＜ 0.05).As the dose of prenantal BaP exprosure increased,the gene expression patterns were downregulated,upregulated,and fluctuated.Twenty-six signaling pathways with differently expressed genes mainly focused on:growth and development,toxicant metabolism and inflammation (P ＜ 0.05).The data from gene network analysis demonstrated that CYP2C13,GSTO1,Rela,MAPK8 and Plcg1 were the key genes in the gene network.Conclusion Gene expression patterns of offsprings' hepatocytes were influenced by prenatal BaP exposure.Some key genes and signal pathways were also found.The study provides an important clue for the toxicity and mechanisms of the prenatal BaP exposure on the growth and development of offspring.