黑龙江立克次体感染对小鼠脾脏免疫相关基因转录的影响

Effect of Rickettsia heilongjiangensis infection on the transcription of immune-related genes in the spleen of mice

目的分析黑龙江立克次体感染小鼠脾脏免疫相关基因的转录变化,筛选影响黑龙江立克次体感染致病进程的关键靶标。方法取20只雌性C3H/HeN小鼠,随机分为未感染组(0 hpi)、感染3 h组(3 hpi)、感染3 d组(3 dpi)及感染6 d组(6 dpi),每组5只,采用尾静脉注射法感染黑龙江立克次体,在4个时相点解剖小鼠取脾,提取总RNA进行测序,筛选出特异性转录基因并进行GO注释、R语言富集及KEGG可视化功能分析。结果小鼠感染黑龙江立克次体后,脾脏有219930个mRNA发生了特异性转录,通过GO注释及富集筛选出显著性变化的3类基因:(1)35个天然免疫相关基因,其中参与γ干扰素(IFN-γ)信号通路的irg1、gbp10基因转录上调最明显(P<0.05);(2)31个获得性免疫相关基因,其中参与细胞凋亡通路的gzmb基因转录上调最明显(P<0.05);(3)41个炎症反应相关基因,其中参与细胞因子与细胞因子受体信号通路的ccl4基因转录上调最明显(P<0.05)。结论宿主IFN-γ信号通路中的irg1、gbp10基因,细胞凋亡通路中的gzmb基因,细胞因子及细胞因子受体信号通路中的ccl4基因影响黑龙江立克次体感染的致病进程。

Objective To screen key targets modulating the progression of Rickettsia heilongjiang infection using a mouse infection model by profiling the transcription expression dynamics of immune-related genes.Methods A total of 20 female C3H/HeN mice were randomized into 4 groups(5 mice/group):0 hpi(hours post-infection),3 hpi,3 dpi(days post-infection),and 6 dpi.Animals were infected with 5×10^(6) CFU Rickettsia heilongjiangensis intravenously.We harvested and extracted total RNA from spleens of animals at the indicated time points.After sequencing the total RNA,we further analyzed the sequencing results by GO annotation,R language enrichment,and KEGG visualization function analysis.Results A total of 19930 mRNAs were specifically transcribed in the spleen post-infection.Using GO annotation and enrichment,we identified three types of immune-related genes with significant changes:first,35 innate immune-related genes,among which the transcription of irg1 and gbp10 genes involved in the IFN-γsignaling pathway rises most significantly(P<0.05);second,31 acquired immune-related genes,among which the transcription of gzmb gene involved in the apoptosis pathway rises most significantly(P<0.05);third,41 inflammatory responses related genes,among which the transcription of ccl4 gene involved in cytokine and cytokine receptor signaling pathway rises most significantly(P<0.05).Conclusions The irg1 and gbp10 genes involved in the IFN-γsignaling pathway,the gzmb gene involved in the apoptosis pathway,and the ccl4 gene involved in the cytokine and cytokine receptor signaling pathway affect the pathogenesis of Rickettsia heilongjiangensis infection.