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目的了解鼻疽诺卡菌感染动物模型小鼠足垫后不同时间的组织病理进展变化以及细胞因子分泌情况,为鼻疽诺卡菌致病机制研究提供依据。方法应用对数生长期的鼻疽诺卡菌注射于小鼠足垫,建立感染模型。将感染1、3、7、14 d后小鼠分别脱臼处死,用手术刀片切取感染小鼠的足垫,制备切片,应用HE染色观察组织不同时间的病理变化,通过免疫组化研究感染过程中IL-6、IL-12、IFN-γ、TNF表达分泌情况。结果通过HE染色观察到感染后第1天中性粒细胞浸润,炎症形成;感染后第3天中性粒细胞脓肿形成,进入急性感染时期;感染后第7天肌肉组织间大量肌纤维母细胞及血管内皮细胞增生,感染转入慢性感染时期;感染后第14天肌肉组织间大量肌纤维母细胞及血管内皮细胞增生,见局灶小脓肿,周围见片状增生的泡沫样巨噬细胞,进入感染修复期。组织免疫组化结果显示,在炎症形成时期,组织中的IL-6和TNF表达水平高于对照组,差异有统计学意义(P<0.05);在急性感染期,组织中仅可见TNF表达水平高于对照组,差异有统计学意义(P<0.05);在慢性感染期,组织中IL-6、IL-12、IFN-γ和TNF表达量均高于对照组,差异有统计学意义(P<0.05);在感染修复期,组织中的IL-6、IFN-γ和TNF表达水平高于对照组,差异有统计学意义(P<0.05)。结论鼻疽诺卡菌感染小鼠足垫后出现明显的病理变化周期,在感染第3天开始进入急性感染时期,感染后第7天转入慢性感染时期,感染后第14天进入组织修复期,并且IL-6、IFN-γ、TNF参与了炎症反应,尤其是IL-6表达量最高,主要促进Th0向Th1和Th17亚群分化,激活巨噬细胞、树突状细胞和NK细胞等,增强其吞噬和杀伤功能,以杀伤胞内鼻疽诺卡菌。
Objective To understand the histopathological changes and secretion of cytokines at different time points after foot pad in nosocomial mice infected with nocardia and provide basis for the pathogenesis of nodularis nocardia. Methods The logarithmic growth stage of Nocardia Nocardia was injected into foot pad of mice to establish an infection model. The mice were sacrificed on days 1, 3, 7, and 14 after infection respectively. The foot pads of infected mice were cut with scalpel blades to prepare sections. The pathological changes at different time points were observed by HE staining and the infection was observed by immunohistochemistry IL-6, IL-12, IFN-γ, TNF expression and secretion. Results The neutrophil infiltration and inflammation were observed on the first day after infection by HE staining. Neutrophilic abscess formed on the third day after infection and entered the period of acute infection. On the 7th day after infection, a large number of myofibroblasts and Vascular endothelial cell hyperplasia, infection into chronic infection; a large number of myofibroblasts and vascular endothelial cell proliferation between the muscle tissue 14 days after infection, see the focal abscess, around the see flaky hyperplasia of foamy macrophages, enter the infection Repair period. Tissue immunohistochemistry results showed that the levels of IL-6 and TNF in the tissue during the period of inflammation were higher than those in the control group (P <0.05); in the acute infection period, only the expression of TNF was found in the tissue (P <0.05). The expression of IL-6, IL-12, IFN-γ and TNF in chronic infection group were higher than those in control group, the difference was statistically significant (P < P <0.05). The levels of IL-6, IFN-γ and TNF in the tissue during infection repair were significantly higher than those in the control group (P <0.05). Conclusions The pathological changes of Nocardia nosocomiatum in footpad of mice appeared obvious pathological change period, and began to enter the acute infection stage on the third day after infection. After infection, the mice were transferred to the chronic infection stage and entered the tissue repair stage on the fourteenth day after infection. , And IL-6, IFN-γ and TNF are involved in the inflammatory reaction, especially the highest expression of IL-6, which mainly promote differentiation of Th0 and Th17 subtypes, activate macrophages, dendritic cells and NK cells, Enhance their phagocytosis and killing function, in order to kill intracellular nocturnal Nocardia.