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目的建立奇异变形杆菌感染人克隆结肠腺癌细胞Caco-2的细胞模型,比较不同来源奇异变形杆菌感染Caco-2细胞的致病性特征。方法活细胞动态观察优化奇异变形杆菌感染Caco-2细胞的感染条件,采用台盼蓝细胞膜通透性检测、稀释平板计数、吉姆萨(Giemsa)染色及透射电子显微镜观察不同来源奇异变形杆菌感染Caco-2细胞后细胞形态学及胞内感染情况的变化,评价该模型中不同来源奇异变形杆菌侵入细胞的特点和病理损伤规律。结果不同来源奇异变形杆菌感染Caco-2细胞的最佳感染复数(MOI)均为100∶1,感染时间均为2 h。致腹泻奇异变形杆菌C02011感染Caco-2细胞的细胞膜通透性改变,黏附率为(0.647±0.038)%,侵袭率为(0.261±0.005)%。健康人群体检株奇异变形杆菌B02005的细胞膜通透性出现轻微受损,黏附率为(0.975±0.030)%,侵袭率为(0.134±0.003)%。尿道分离奇异变形杆菌标准株ATCC29906的黏附率为(0.303±0.027)%,侵袭率为(0.104±0.003)%,并未发现Caco-2的细胞膜发生变化。吉姆萨染色和透射电子显微镜观察与上述方法结果一致,说明了不同来源奇异变形杆菌感染Caco-2细胞的致病性不同(P<0.05);同时,透射电子显微镜实验也验证了细胞模型的成功建立。结论成功构建奇异变形杆菌感染Caco-2的细胞模型,并分析了不同来源奇异变形杆菌的致病性特征。
Objective To establish a Caco-2 cell model of human colon adenocarcinoma cells infected by Proteus mirabilis and to compare the pathogenicity of Caco-2 cells infected by different sources of Proteus mirabilis. Methods The living conditions of Caco-2 cells infected by Proteus mirabilis were optimized by dynamic observation of living cells. Trypan blue cell membrane permeability assay, dilution plate count, Giemsa staining and transmission electron microscopy were used to observe the infection of Caco-2 -2 cell morphology and intracellular infection changes in the model to evaluate the different sources of Proteus mirabilis invasive characteristics of cells and pathological damage. Results The optimal multiplicity of infection (MOI) of Caco-2 cells from different sources of Proteus mirabilis was 100: 1 and the infection time was 2 h. The cell membrane permeability of Caco-2 cells infected with Proteus mirabilis C02011 was (0.647 ± 0.038)% and the invasion rate was (0.261 ± 0.005)%. The susceptibility of Proteus mirabilis B02005 was slightly impaired. The adhesion rate was (0.975 ± 0.030)% and the invasion rate was (0.134 ± 0.003)%. Urethral isolation of Proteus mirabilis ATCC29906 standard strain adhesion rate was (0.303 ± 0.027)%, the invasion rate was (0.104 ± 0.003)%, did not find Caco-2 cell membrane changes. The results of Giemsa staining and transmission electron microscopy were consistent with those of the above methods, indicating that the pathogenicity of Caco-2 cells infected by different sources of Proteus mirabilis was different (P <0.05). Transmission electron microscopy also verified the success of the cell model set up. Conclusion The cell model of Caco-2 infected by Proteus mirabilis was successfully constructed and the pathogenicity of Proteus mirabilis from different sources was analyzed.