CAJ | 학술논문

目的：DON致小鼠骨骼畸形的分子基础。方法：取孕期小鼠30只，随机分成3个不同浓度DON染毒组、溶剂对照组和空白对照组，每组6只，将DON染毒组孕鼠于孕期第7～10天连续腹腔注射DON，染毒后第18天麻醉剖腹取出胎鼠骨骼组织，提取椎骨骨骼组织总RNA，通过反转录法获取畸形骨骼组织和正常对照组骨骼组织的cDNA，并对其分别进行荧光标记，采用全基因组芯片技术对小鼠胚胎畸形骨骼组织与正常对照组织的差异基因表达谱进行分析，采用基因本体学分析软件对差异表达基因进行功能分析， DAVID database数据库筛选差异表达基因涉及的信号通路，并利用荧光定量PCR(qPCR)技术对基因芯片结果进行验证。结果：在DON染毒组小鼠全基因组芯片上筛选出差异基因282个，其中下调148个，上调134个；在差异表达基因分析的基础上，发现差异表达基因涉及到的通路有153个。qPCR结果表明，上调基因Fbn1 Co19a2 Papss2 Pax1F和下调基因Runx2 Pthlh等6个差异表达基因的相对定量结果与基因芯片检验结果表达趋势一致。结论：应用基因表达谱芯片初步筛选出DON所致胎鼠畸形骨骼与正常胎鼠骨骼组织中的差异表达基因，并通过通路分析发现，上述差异表达基因涉及多条信号传导通路。筛选脱氧雪腐镰刀菌烯醇(DON)致小鼠胚胎畸形骨骼组织的差异表达基因及相关信号传导通路，从基因水平上阐明2014-05-11；修订日期：2014-01-06
목적：DON치소서골격기형적분자기출。방법：취잉기소서30지，수궤분성3개불동농도DON염독조、용제대조조화공백대조조，매조6지，장DON염독조잉서우잉기제7～10천련속복강주사DON，염독후제18천마취부복취출태서골격조직，제취추골골격조직총RNA，통과반전록법획취기형골격조직화정상대조조골격조직적cDNA，병대기분별진행형광표기，채용전기인조심편기술대소서배태기형골격조직여정상대조조직적차이기인표체보진행분석，채용기인본체학분석연건대차이표체기인진행공능분석， DAVID database수거고사선차이표체기인섭급적신호통로，병이용형광정량PCR(qPCR)기술대기인심편결과진행험증。결과：재DON염독조소서전기인조심편상사선출차이기인282개，기중하조148개，상조134개；재차이표체기인분석적기출상，발현차이표체기인섭급도적통로유153개。qPCR결과표명，상조기인Fbn1 Co19a2 Papss2 Pax1F화하조기인Runx2 Pthlh등6개차이표체기인적상대정량결과여기인심편검험결과표체추세일치。결론：응용기인표체보심편초보사선출DON소치태서기형골격여정상태서골격조직중적차이표체기인，병통과통로분석발현，상술차이표체기인섭급다조신호전도통로。사선탈양설부렴도균희순(DON)치소서배태기형골격조직적차이표체기인급상관신호전도통로，종기인수평상천명2014-05-11；수정일기：2014-01-06
OBJECTIVE:To screen the differential gene expression and related pathways of fetal skeletal malformations induced by deoxynivalenol(DON) in mice,and to explore the mechanism of deformities induced by DON at the molecular level.METHODS:30 pregnant mice were randomly divided into 3 experimental groups and 2 control groups,6 in each group. DON injection was performed from days 7 to 10 of pregnancy by intraperitoneal injection into pregnant mice. At GD 18,all mice were killed under isoflurane anesthesia,and vertebral bone tissue of fetuses were collected. Total RNA of vertebral bone tissues was extracted and cDNA was obtained by RT-PCR. Using whole genome microarray technology,the gene expression profiles and the pathways of the rat vertebral bone tissue were studied. Analysis of the function of differentially expressed genes was performed by using software of gene ontology. Screening signaling pathways of differentially expressed genes was done by DAVID database. The results were verified by fluorescence quantitative PCR(qPCR) technology. RESULTS:Microarray analysis showed that 282 genes,including 148 down-regulated and 134 up-regulated genes,were abnormally expressed in fetal vertebral bones after maternal DON exposure. 153 pathways were related to the differentially expressed genes. The relative quantitative results ofqPCR were consistent w ith t he r esults o f g ene c hip t est i nFbn1,Co19a2,Papss2,Pax1,Runx2 a ndPthlh g enes.CONCLUSION: There were differential gene expressions in deformed fetal skeleton between deoxynivalenol-treated rats and normal rats, involving multiple signaling pathways. The differentially expressed genes and signaling pathways may be related to the molecular mechanisms of deformities induced by DON.