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分析感染奇异变形杆菌和聚集性大肠埃希菌(EAEC)菌株的 BALB/c 小鼠尿路炎症细胞因子的表达。

Analysis of inflammatory cytokine expression in the urinary tract of BALB/c mice infected with Proteus (P.) mirabilis and enteroaggregative Escherichia (E.) coli (EAEC) strains.

机构信息

Maestria en Ciencias Farmaceuticas, Universidad Autonoma Metropolitana Unidad Xochimilco, Calzada del Hueso No. 1100, CP 04960, CDMX, Mexico.

Departamento de Sistemas Biologicos, Universidad Autonoma Metropolitana Unidad Xochimilco, Calzada del Hueso No. 1100, CP 04960, CDMX, Mexico.

出版信息

Folia Microbiol (Praha). 2020 Feb;65(1):133-142. doi: 10.1007/s12223-019-00714-2. Epub 2019 May 19.

Abstract

This study aimed to analyze the proinflammatory cytokine mRNA expression in the urinary tract of BALB/c mice infected with bacterial strains with uropathogenic potential. Groups of four 6-week-old female BALB/c mice were intraurethrally inoculated with 5 × 10 colony-forming units (CFU) of P. mirabilis ATCC29906, EAEC O42, P. mirabilis RTX339, or sterile saline (control group) and then sacrificed at 0, 2, 4, 7, or 10 days post-infection (p.i.). Samples were cultured to determine the CFU/mL in urine or CFU/g in the bladders and kidneys. Cytokine expression (tumor necrosis factor (TNF)-α and interleukin (IL)-1β, -6, and -8) was evaluated in the target organs using real-time PCR and immunohistochemistry; histology was examined with hematoxylin and eosin staining. The results are presented as the means and standard deviations and were compared using one-way ANOVA, with p < 0.05 indicating significant differences. Bacteriuria was not detected in the infected groups; bacterial colonization occurred in the target organs at all time points, but was higher in mice infected with EAEC O42 or P. mirabilis RTX339 at 7 days p.i. The expression of all cytokine mRNAs was seen, but only the levels of the IL-8 protein increased in situ at 7 days p.i. in the P. mirabilis RTX339 and EAEC O42 groups in both organs. Morphological alterations, observed in all of the infected groups, were more prominent in the EAEC O42 and P. mirabilis RTX339 groups. The findings provide insights into the uropathogenicity and inflammatory cytokine expression in the urinary tract of mice infected with three previously untested bacterial strains.

摘要

本研究旨在分析具有泌尿道致病性的细菌菌株感染 BALB/c 小鼠尿路的促炎细胞因子 mRNA 表达。将 4 组每组 4 只 6 周龄雌性 BALB/c 小鼠经尿道接种 5×10 个菌落形成单位(CFU)的 P. mirabilis ATCC29906、EAEC O42、P. mirabilis RTX339 或无菌生理盐水(对照组),然后于感染后 0、2、4、7 或 10 天处死。采集尿液样本以确定 CFU/mL,采集膀胱和肾脏样本以确定 CFU/g。使用实时 PCR 和免疫组织化学评估靶器官中的细胞因子表达(肿瘤坏死因子(TNF)-α 和白细胞介素(IL)-1β、-6 和 -8);使用苏木精和伊红染色检查组织学。结果以平均值和标准差表示,并使用单向方差分析进行比较,p<0.05 表示差异有统计学意义。感染组未检测到菌尿;在所有时间点,细菌均定植于靶器官,但在感染 EAEC O42 或 P. mirabilis RTX339 的小鼠中,7 天 p.i. 时定植水平更高。所有细胞因子 mRNA 的表达均可见,但仅在 P. mirabilis RTX339 和 EAEC O42 组的 7 天 p.i. 时在两个器官中观察到原位 IL-8 蛋白水平增加。在所有感染组中观察到的形态改变在 EAEC O42 和 P. mirabilis RTX339 组更为明显。这些发现深入了解了三种未经测试的细菌菌株感染小鼠尿路的尿路致病性和炎症细胞因子表达。

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