犬弓首蛔虫感染比格犬不同阶段肝circRNAs表达模式的分析

doi:10.11843/j.issn.0366-6964.2021.012.019

摘要/Abstract

摘要： 旨在探究终末宿主肝环状RNA （circRNAs）在犬弓首蛔虫诱导致病过程中的潜在调控作用。18只6~7周龄的比格犬被平均分为3组（感染后0.5 d组、感染后1 d组和感染后36 d组），每组包括感染和对照各3只，分别于感染后对应的时间点收集肝样品。提取感染和对照组肝的总RNA，利用高通量RNA测序，构建circRNA文库，对差异表达的circRNAs进行分析，并对差异表达circRNAs的亲本基因进行GO注释和KEGG富集分析。随机选取9个差异表达circRNAs进行实时荧光定量PCR验证。结果显示，与对照组相比，在感染后0.5、1和36 d分别有94、103和84个犬肝的差异表达circRNAs。Venn图显示，3个感染阶段没有共同表达的差异circRNAs；GO和KEGG分析显示，差异表达的circRNAs参与了宿主肝免疫与炎症的相关通路，其中差异表达的circRNA novel_circ_0016108、novel_circ_0016184和novel_circ_0027468与犬弓首蛔虫引起的天然免疫相关；novel_circ_0002212参与了犬弓首蛔虫引起的肝炎症反应；novel_circ_0002212和novel_circ_0019907在宿主肝抵抗犬弓首蛔虫侵袭的过程中发挥作用。RT-qPCR验证结果显示大多数差异表达的circRNAs与高通量测序结果一致。结果表明，肝circRNAs在犬弓首蛔虫感染终末宿主的过程中发挥着重要作用，这为进一步探究犬弓首蛔虫与宿主之间的相互作用提供了新的信息，并且也为促进疾病干预措施的制定提供参考。

关键词: 犬弓首蛔虫, 比格犬, 肝, RNA测序, 环状RNA

Abstract: This study was conducted to investigate the potential regulatory role of circRNAs in the definitive host liver in the pathogenesis induced by Toxocara canis. The six-to-seven-week-old Beagle puppies (n=18) were averagely divided into three stages (0.5 day post infection, 1 dpi, and 36 dpi), and their liver samples were collected at the corresponding time points after infection. Each stage included infected group and control group. Total RNA was extracted from the livers of infected and uninfected puppies, and the circRNA libraries were constructed and sequenced by high-throughput RNA sequencing. The differentially expressed (DE) circRNAs were analyzed, and GO and KEGG enrichment analysis were performed using the source genes of DE circRNAs. The nine DEcircRNAs were randomly selected for quantitative real-time PCR (RT-qPCR) verification. Compared to the control groups, 94, 103, and 84 DE circRNAs of dogs' livers were detected in the livers of infected dogs at the three infection stages, respectively. Venn plot analysis showed that there were no co-expressed DE circRNAs among the three stages of infection. GO and KEGG analysis showed that some DE circRNAs were related to immunity and inflammation in the host liver. Among these DE circRNAs, novel_circ_0016108, novel_circ_0016184 and novel_circ_0027468 were related to the natural immunity of host against T. canis infection; novel_circ_0002212 was involved in liver inflammation caused by T. canis; novel_circ_0002212 and novel_circ_0019907 play a role in the host liver's resistance to T. canis invasion. RT-qPCR validation showed that the expression trends of DE circRNAs obtained by both methods were consistent. These results suggest that circRNAs play an important role in the infection of definitive host (dogs) by T. canis. This study provided new information for further understanding of the interaction between T. canis and its definitive host, and will also facilitate the development of interventions for the disease.

Key words: Toxocara canis, Beagle dogs, liver, RNA sequencing, circRNAs

中图分类号:

S855.9

邹扬, 郑文斌, 张金鹏, 路义鑫, 朱兴全. 犬弓首蛔虫感染比格犬不同阶段肝circRNAs表达模式的分析[J]. 畜牧兽医学报, 2021, 52(12): 3524-3534.

ZOU Yang, ZHENG Wenbin, ZHANG Jinpeng, LU Yixin, ZHU Xingquan. CircRNAs Expression Patterns in the Liver of Beagle Dogs at Different Stages of Toxocara canis Infection[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(12): 3524-3534.

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