摘要
旨在揭示内质网应激(ERS)标记基因Grp78是否参与鹅肥肝中炎症/免疫相关基因的表达调控。本试验选择健康的、体重相近的70日龄朗德鹅30只(未分公母),随机分为对照组(n=15)和填饲组(n=15)。利用荧光定量PCR测定不同填饲阶段(填饲7、14、19 d)不同组织中Grp78的相对表达量(n=4),以明晰Grp78与鹅肥肝形成的关联;在鹅原代肝细胞中过表达Grp78,并通过RNA-seq分析Grp78所影响的信号通路和基因(n=3);利用荧光定量PCR检测鹅肥肝中与炎症/免疫相关的基因表达量(n=4),以明晰这些基因与Grp78的关联。结果表明,肝和腹脂中Grp78的表达量受填饲影响(肝中下降,而腹脂中上升)。Grp78基因的过表达试验表明,ERS除了可影响已知的代谢病和细胞凋亡通路外,还影响信号转导、免疫等通路。在免疫/炎症方面,"Toll-like receptor signaling pathway"、"TNFαsignaling pathway"、"NF-kappa B signaling pathway"等通路最为突出。此外,免疫/炎症相关基因Tnfsf10、Map3k7cl在鹅原代细胞中的表达受Grp78过表达影响,在鹅肥肝中受填饲影响。总之,本研究揭示了Grp78/ERS新的生物学功能,即对细胞的免疫/炎症状态进行调控,并借此参与鹅肥肝的形成。
The aim of this study was to investigate whether endoplasmic reticulum stress(ERS)marker gene Grp 78 was involved in regulation on the expression of inflammation/immunity-related genes in goose fatty liver.Thirty healthy 70-day-old Landaise geese(male or female)with similar body weight were randomly divided into the control group(n=15)and the overfeeding group(n=15).The relative expression of Grp 78 in different tissues at different overfeeding periods(7,14,19 d)was determined by fluorescent quantitative PCR to clarify the relationship between Grp 78 and geese fatty liver formation;Grp 78 was overexpressed in goose primary hepatocytes,and the signaling pathways and genes affected by Grp 78 were analyzed by RNA-seq(n=3);The expression of genes related to inflammation/immunity was determined in goose fatty liver by fluorescent quantitative PCR(n=4)to clarify the association of these genes with Grp 78.The results showed that the expression of Grp 78 in the liver and abdominal fat was affected by overfeeding(decreased in liver and increased in abdominal fat).The result of overexpression of Grp 78 gene suggested that ERS not only participated in the known metabolic diseases and cell apoptosis pathways,but also had an effect on signal transduction and immune pathways.The Toll-like receptor signaling pathway,TNFαsignaling pathway and NF-kappa B signaling pathway were the most prominent ones in regard to immunity/inflammation.In addition,the expression of immune/inflammation related genes,Tnfsf 10 and Map 3k7cl,was affected by Grp 78 overexpression in the goose primary hepatocytes and was affected by overfeeding in goose fatty liver.In summary,this study revealed a new biological function of Grp 78/ERS,that is,to regulate cellular immune/inflammatory state and participate in the formation of goose fatty liver.
作者
刘同君
赵盼
赵敏孟
刘龙
崔恒宓
龚道清
耿拓宇
LIU Tongjun;ZHAO Pan;ZHAO Minmeng;LIU Long;CUI Hengmi;GONG Daoqing;GENG Tuoyu(College of Animal Science and Technology,Yangzhou University,Yangzhou 225009,China)
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2019年第4期727-737,共11页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家自然科学基金面上项目(31472086)
江苏高校优势学科建设工程资助项目(PAPD)