摘要
生长发育过程中鸡胚不同组织的抗氧化酶活性是不断变化的,在整个鸡胚孵化期间超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)对于抵抗活性氧起着至关重要的作用。为探究调控鸡胚肝组织抗氧化相关的关键基因及其作用机制,文章通过检测鸡胚12~18 d肝脏组织中的SOD和GSH-Px的酶活变化,选取具有极高和极低抗氧化值的个体进行后期的转录组测序,结果表明:12~18 d肝脏组织中的SOD和GSH-Px的酶活均呈现先降低后增高的趋势,同时12、16 d鸡胚肝脏组织SOD酶活和GSH-Px酶活差异达到显著水平(P<0.01),基于此,对12、16 d的肝脏组织进行转录组测序,筛选肝脏组织中差异表达的基因,并对其进行了富集分析筛选关键作用通路。转录组共发现显著差异表达基因216个(其中上调基因81个,下调基因135个),结合基因功能、GO富集分析和KEGG富集分析鉴定出GSTA3、GSTA、TTPA、MGST3、HMOX2、GSTA1、ODC1、MAOA和GST可能是影响鸡胚发育过程中抗氧化酶活性变化的关键候选基因。另外,随机挑选5个显著差异基因进行实时荧光定量聚合酶链式反应(polymerase chain reaction,PCR)测定,鉴定结果与转录组测序结果一致,从而验证了测序数据的准确性。该研究鉴定的新的鸡胚抗氧化相关基因为进一步探究鸡胚肝脏的抗氧化机制奠定了一定的理论基础。
During growth and development,antioxidant enzyme activities in different tissues of chicken embryos are constantly changing.Of these,superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)play a vital role in resisting reactive oxygen species(ROS),during the development of the embryo.To detect the key genes related to the regulation of antioxidation in chicken embryo liver tissues,the enzyme activity of SOD and GSH-Px in liver tissue from 12 d to 18 d of chicken embryos was detected and the individuals with extremely high and low antioxidant values were chosen for subsequent transcriptome sequencing.The results showed that from 12 d to 18 d the activities of SOD and GSH-Px in liver tissues decreased first and then increased,while the activity of SOD and GSH-Px at 12 d and 16 d had extremely significant differences in antioxidant value(P<0.01).Based on these phenotype results,the RNA-seq was used to detect the differentially expressed genes in the liver tissues of chicken embryos and some key pathways for enrichment analysis were explored.The results showed that there are 216 significantly differentially expressed genes(81 up-and 135 down-regulated),GO enrichment analysis and KEGG enrichment analysis showed that GSTA3,GSTA,TTPA,MGST3,HMOX2,GSTA1,ODC1,MAOA,GST may be the key candidate genes influencing the antioxidant enzyme activity associated with chicken embryo development.Additionally,five significantly differentially expressed genes were randomly selected from the sequencing results for reverse transcription-polymerase chain reaction(RT-PCR)to verify the accuracy of the sequencing data,and the RT-PCR results were consistent with the results of RNA-seq.All results may have a good effect on exploring the antioxidant mechanism in liver tissues of chicken embryos.
作者
李春晖
何程实
吴建鑫
韦张其
杨少华
刘国庆
LI Chunhui;HE Chengshi;WU Jianxin;WEI Zhangqi;YANG Shaohua;LIU Guoqing(School of Food and Biological Engineering, Hefei University of Technology, Hefei 230601, China)
出处
《合肥工业大学学报（自然科学版）》
CAS
北大核心
2021年第10期1430-1436,共7页
Journal of Hefei University of Technology：Natural Science
基金
安徽省科技攻关计划资助项目(1501031092)。
关键词
鸡胚
肝脏组织
转录组测序
差异表达基因
抗氧化
chicken embryo
liver tissue
transcriptome sequencing
differentially expressed genes
antioxidation
