日本血吸虫感染小鼠肝脏枯否细胞差异表达MicroRNA及基因的分析

Differently expressed micro RNAs and genes in hepatic Kupffer cells after Schistosoma japonicum infection

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 identifiy differently expressed micro RNAs（mi RNAs）and m RNAs in hepaticKupffer cells（KCs） after Sc^（hi）stosoma japonicum（S. japonicum） infection. Methods Mice were divided into control group andinfection group randomly. F4/80^（hi）CD11 b^（lo）KCs were isolated by gradient elution and f^（lo）w cytometry from normal and S.japonicum-infected mice at 6 weeks post infection. Then, mi RNA microarray and RNA-seq were performed. Real-timefluorescent quantification PCR was used to validate differential expression of mi RNAs and m RNAs. Results Compared tocontrol group, after infection with S. japonicum, the number of F4/80^（hi）CD11 b^（lo）KCs was decreased quickly. Meanw^（hi）le,F4/80^（lo）CD11b^（hi）non-parenchymal cells（NPCs） was generated in liver. The F4/80^（hi）CD11 b^（lo）KCs isolated from mice in controlgroup and the infection group at 6 weeks post infection were analyzed by mi RNA microarray and RNA-seq of RNA. The datashowed that 106 mi RNAs and 1 083 m RNAs including genes encoding various chemokines were differentially expressed afterinfection. Conclusions S. japonicum infection could result in the decrease of F4/80^（hi）CD11 b^（lo）KCs in liver. A large number ofmi RNAs and m RNAs that were differentially expressed in F4/80^（hi）CD11 b^（lo）KCs after S. japonicum infection were identified.