Th1/Th2型免疫反应相关基因在巨噬细胞RAW 264.7应对细粒棘球绦虫原头蚴刺激时的表达谱研究

本研究旨在解析巨噬细胞RAW264.7在应对细粒棘球绦虫原头蚴刺激时其Th1、Th2型免疫反应相关基因的差异表达规律,为进一步揭示巨噬细胞抗细粒棘球绦虫原头蚴免疫调控机制奠定理论基础.将细粒棘球绦虫原头蚴和巨噬细胞RAW264.7共培养6、24、72 h,收集RAW264.7细胞,提取总RNA,构建cDNA文库,利用R...

Study on the Expression Profile of Th1/Th2 Immune Response-related Genes in Macrophage RAW264.7 in Response to the Stimulation of Echinococcus granulosus Protoscoleces

This study aimed to analyze the differential expression of Th1 and Th2 type immune response-related genes in macrophages RAW264.7 in response to Echinococcus granulosus protoscoleces stimulation,this study lays a theoretical foundation for further revealing the immune regulation mechanism of macrophages against protoscoleces.The protoscoleces and macrophage RAW264.7 were cultured for 6,24 and 72 hours,total RNA was extracted from RAW264.7 cells,and cDNA library was constructed.The results showed that when the protoscoleces(PSC)were cultured with macrophages RAW264.7 for 6,24,and 72 hours,compared with the PBS control group,there were 848,3745 and 7009 genes in PSC treatment group showed significant difference,in which 415,1159 and 2237 genes were up-regulated and 433,2586 and 4772 genes were down-regulated respectively.The results of differential expression analysis of Th1 and Th2 immune response-related genes showed that when macrophages RAW264.7 were co-cultured with protoscoleces for 6 hours,a total of 31 Th1 and Th2 type immune response-related genes showed significant changes in expression,including 15 genes significantly up-regulated,16 significantly down-regulated.When macrophages RAW264.7 were cultured with protoscoleces for 24 hours,there were 112 of Th1 and Th2 type immune response-related genes were significantly changed,of which 34 were up-regulated and 78 down-regulated.When macrophages RAW264.7 were cultured with protoscoleces for 72 hours,the expression of 212 Th1 and Th2 type immune response related genes changed significantly,among which 50 genes were up-regulated and 162 genes were down-regulated.These differentially expressed genes include the Leucine-rich repeat protein LRRCs,the G protein coupled receptor GPCRs,the C type Lectin receptor CLRs,the Scavenger receptor SRS,and some downstream molecules of PRR.At the same time,the results showed that the expression of Th1-related genes such as Tnfrsf1a,Ifnar1 Tnfrsf12a and so on was significantly up-regulated when the protoscoleces were cultured with macrophages RAW264.7 for 6 and 24 hours.However,when the protoscoleces were cultured with macrophages RAW264.7 for 72 hours,the expression of Th2-related genes such as IL4 and IL6 was significantly up-regulated.At the same time,some differentially expressed genes were selected at random and verified by qRT-PCR.The results showed that the expression trend was consistent with that of RNA-seq.To sum up,this study systematically analyzed the different expression profiles of genes related to macrophages RAW264.7 responding to protoscoleces stimulation in different time periods,the genes associated with macrophage RAW264.7 response to protoscoleces stimulation and innate immunity were screened,this study provides a basis for further understanding the role of these differentially expressed genes in response to protoscoleces stimulation by macrophages RAW264.7 and their regulatory mechanisms.