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目的本研究在建立免疫抑制小鼠内毒素血症模型的基础上,观察机体肺损伤的现象以及发生机制。方法免疫抑制小鼠内毒素血症模型建立,选取Blb/c小鼠,腹腔内注射甲基强的松龙30 mg/kg连续3 d,随后静脉注射内毒素,10 mg/kg剂量,于注射后26、h时点取血浆,在此基础上,50只小鼠分为免疫抑制内毒素血症模型组(模型组)以及甲基强的松龙对照组(激素组)与空白对照组,检测指标分别:①肺部病理学改变,采用HE染色;②肺组织细胞因子表达检测:分别用免疫组化与ELISA检测肺组织中TNF-α与IL-10表达水平;采用免疫组化检测肺组织诱导型一氧化氮合酶(iNOS)表达情况,肺组织一氧化氮(NO)水平采用化学比色法检测;③是采用化学比色法与ELISA检测血浆一氧化氮、TNF-α、INF-γ、IL-10等细胞因子水平。结果免疫抑制小鼠内毒素血症模型小鼠出现急性肺损伤的病理改变,TNF-α、IL-10与iNOS在肺间质浸润的炎性细胞中阳性表达;在2 h与6 h时点,肺组织与血浆TNF-α与NO的水平明显高于激素组与空白组(P<0.05);在6 h时点,模型组小鼠肺组织与血浆IL-10的水平明显高于激素组与空白组(P<0.05)。结论免疫抑制内毒素血症模型小鼠急性肺损伤与机体巨噬细胞过度活化而释放大量炎性细胞因子有关。
OBJECTIVE: To establish a model of endotoxemia in immunosuppressed mice to observe the pathogenesis and mechanism of lung injury. Methods Immunosuppressive mouse model of endotoxemia was established. Blb / c mice were injected intraperitoneally with methylprednisolone 30 mg / kg for 3 consecutive days, followed by intravenous injection of endotoxin at a dose of 10 mg / kg. After injection After 26, h, the plasma was taken at the point of time. On this basis, 50 mice were divided into immunosuppressed endotoxemia model group (model group), methylprednisolone control group (hormone group) and blank control group, The detection indexes were: ① pathological changes of lungs, HE staining; ②the detection of cytokines in lung tissue: the expression of TNF-α and IL-10 in lung tissues were detected by immunohistochemistry and ELISA respectively; The expression of tissue-inducible nitric oxide synthase (iNOS) and the level of nitric oxide (NO) in lung tissue were detected by chemical colorimetry. ③The levels of plasma nitric oxide, TNF-α, INF -γ, IL-10 and other cytokines levels. Results The pathological changes of acute lung injury in immunosuppressed mice with endotoxemia were observed. The expression of TNF-α, IL-10 and iNOS were positive in inflammatory cells infiltrating lung interstitium. At 2 h and 6 h , And the level of TNF-α and NO in lung tissue and plasma was significantly higher than that in the hormone group and the blank group (P <0.05). At 6 h, the level of IL-10 in the lung tissue and plasma in the model group was significantly higher than that in the hormone group And blank group (P <0.05). Conclusion The acute lung injury induced by immunosuppressed endotoxemia mice is related to the excessive activation of macrophages and the release of a large number of inflammatory cytokines.