摘要
采用腹腔注射的方式构建小鼠感染多杀性巴氏杆菌(Pasteurella multitocida,Pm)HN01模型,通过半数致死量(LD_(50))、组织载菌量和组织病理学变化评估Pm HN01的毒力。对小鼠肺脏进行转录组高通量测序,并利用qPCR、Western blot和免疫组织化学方法验证测序结果。结果表明,Pm HN01主要引起小鼠肺脏和脾脏的充血及基础结构的损伤。转录组测序结果显示,Pm HN01感染引起小鼠肺脏1465个基因显著差异上调,1096个基因显著差异下调,差异表达基因显著富集于细胞因子-细胞因子受体互作通路、癌症通路、IL-17信号通路等多条免疫相关通路。随机选择10个与免疫相关的差异表达基因进行qPCR验证,结果显示其中9个基因表达趋势与转录组测序一致。Western blot和免疫组化结果表明,感染小鼠肺脏的SAA3蛋白表达量显著升高,并且主要分布于支气管腔内壁。本研究对Pm HN01的毒力进行了初步评估,通过转录组数据揭示了小鼠在感染Pm HN01后肺脏的基因表达差异,筛选出可能在抗Pm HN01感染中发挥重要功能的免疫通路和关键蛋白,为进一步研究Pm HN01的致病机制和宿主免疫应答提供参考。
The experiment established a mouse model infected with Pasteurella multocida(Pm)HN01 intraperitoneally,evaluated the virulence of Pm HN01 through median lethal dose,tissue bacterial load,and histopathological changes.The mouse lungs were then used for transcriptome sequencing.qPCR,western blot,and immunohistochemistry methods were used for verifying the sequencing results.The results showed that Pm HN01 mainly caused congestion and structural damage of the lung and spleen of mice.Transcriptome sequencing results showed that there were total of 1465 up-regulated genes and 1096 down-regulated genes in mouse lungs caused by Pm HN01 infection.Differentially expressed genes were significantly enriched in immune-related pathways such as cytokine-cytokine receptor interaction,cancer pathway,and IL-17 signaling pathway etc.Ten differentially expressed genes related to immunity were randomly selected and verified by qPCR and the results showed that expression trends of the nine genes were consistent with transcriptome sequencing.Western blot and immunohistochemistry confirmed the expression of SAA3 protein was significantly increased and mainly distributed in the inner wall of bronchial lumen in infected mouse lungs.This study provided a preliminary assessment of the virulence of Pm HN01,revealed gene expression pattern in mouse lungs after Pm infection through transcriptome data,screened out immune pathways and key important proteins that may play crucial roles in anti-Pm infection,and offered theoretical basis for further research on pathogenic mechanisms of Pm and host immune responses.
作者
程逸文
张振兴
李昊洋
何美荣
陈思
满初日嘎
杜丽
王凤阳
陈巧玲
CHENG Yi-wen;ZHANG Zhen-xing;LI Hao-yang;HE Mei-rong;CHEN Si;MAN Chu-ri-ga;DU Li;WANG Feng-yang;CHEN Qiao-ling(College of Tropical Agriculture and Forestry,Hainan Key Laboratory of Tropical Animal Reproduction and Breeding and Epidemic Disease Research,Hainan University,Haikou 570228)
出处
《微生物学杂志》
北大核心
2025年第6期74-83,共10页
Journal of Microbiology
基金
海南省自然科学基金项目(324RC447)
海南省院士创新平台科研专项(YSPTZX202013)。
