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目的筛选并验证日本血吸虫感染后,肝脏枯否细胞(Kupffer cells,KCs)内差异表达micro RNAs(mi RNAs)及基因。方法实验小鼠随机分为对照组、感染日本血吸虫组。用密度梯度离心和流式分选相结合的方法,分离肝脏F4/80~(hi)CD11b~(lo)枯否细胞。取感染后6周样本进行mi RNA芯片和转录组测序分析,并利用实时荧光定量PCR方法进行验证。结果与对照组相比,感染组肝脏F4/80~(hi)CD11b~(lo)枯否细胞数量急剧减少,与此同时感染组肝脏逐渐出现F4/80loCD11bhi非实质细胞。采用感染后6周宿主肝脏F4/80~(hi)CD11b~(lo)枯否细胞样本进行mi RNA芯片和转录组测序分析,与对照组相比,感染组肝脏枯否细胞差异表达的mi RNAs有106条,差异表达基因为1 083个,其中包含多个趋化因子。结论日本血吸虫感染导致宿主肝脏F4/80~(hi)CD11b~(lo)枯否细胞数量显著减少,并鉴定一批差异表达的mi RNAs和基因。
Objective To screen and validate the differential expression of miRNAs and genes in Kupffer cells (KCs) after Schistosoma japonicum infection. Methods Experimental mice were randomly divided into control group and infected with Schistosoma japonicum group. The hepatic F4 / 80 ~ (hi) CD11b ~ (io) Kupffer cells were isolated by a combination of density gradient centrifugation and flow cytometry. Six weeks after infection, the samples were sequenced and analyzed by real-time PCR. Results Compared with the control group, the number of F4 / 80 ~ (h) CD11b ~ (lo) Kupffer cells in the infected group decreased sharply, while the F4 / 80loCD11bhi non-parenchymal cells gradually appeared in the infected group. The miRNAs and transcriptome of F4 / 80 ~ (h) CD11b ~ (lo) Kupffer cells in host liver were sequenced at 6 weeks after infection. Compared with the control group, miRNAs differentially expressed in liver Kupffer cells There are 106, the number of differentially expressed genes is 1 083, which contains multiple chemokines. Conclusion The infection of Schistosoma japonicum causes a significant decrease in the number of F4 / 80 ~ (h) CD11b ~ (lo) Kupffer cells in the liver and identifies a number of differentially expressed miRNAs and genes.