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目的 探讨脂多糖诱发急性肺炎时 ,大白鼠中性粒细胞碱性磷酸酶 (alkaline phosphatase,AL P)的超微定位及其在炎症过程中的相关功能。 方法 采用腹腔内 (intraperitoneal,IP)注射及气管内 (intratracheal,IT)滴注脂多糖 (lipopolysaccharide,L PS,一种提自大肠杆菌的内毒素 )的方式诱发大白鼠急性肺炎 ,使中性粒细胞在肺内不同部位集聚。用铈作为捕捉剂 ,以检测 AL P活性。对照组为介质中除去底物或加入 2 m mol/L左旋咪唑。电镜下行酶活性定位观察。 结果 IP注射主要导致中性粒细胞在鼠肺肺泡隔毛细血管内集聚 ,而 IT注射多致中性粒细胞聚集于肺泡腔中。在偶见的非炎区肺内中性粒细胞 ,AL P活性产物主要以小球状或管状的膜包颗粒存在于胞质中。L PS腹腔内注射组中 ,AL P活性除出现于胞质内 ,还呈现于中性粒细胞的质膜表面 ;在气管内给药组 ,与毛细血管内皮细胞或肺泡上皮细胞相接触的中性粒细胞质膜表面 AL P活性有所增强 ,而肺泡腔的中性粒细胞质膜表面 AL P活性减弱。在各组标本中 ,对照组均为 AL P阴性。 结论 大鼠肺中性粒细胞 AL P活性可经 L PS注射而增强 ,其活性可能和细胞与细胞间的粘连、细胞外环境以及与细胞的功能状态有关。
Objective To investigate the ultrastructural localization of alkaline phosphatase (ALP) in rat neutrophils induced by lipopolysaccharide (LPS) and its related functions during inflammatory process. Methods Acute pneumonia was induced in rats by intraperitoneal (IP) injection and intratracheal (IT) instillation of lipopolysaccharide (LPS), an endotoxin derived from Escherichia coli. The neutrophils Cells accumulate in different parts of the lung. Cerium was used as a capture reagent to detect ALP activity. Control group to remove the substrate or add 2 m mol / L levamisole. Electron microscopy under the position of enzyme activity. Results IP injection mainly resulted in the agglutination of neutrophils in rat pulmonary alveolar capillaries, whereas IT injection-induced neutrophil accumulation in alveolar spaces. In the occasional non-inflammatory zone of lung neutrophils, the ALP-active product is mainly present in the cytoplasm as small globular or tubular membrane-bound particles. L PS intraperitoneal injection group, AL P activity in addition to appear in the cytoplasm, also presented in the plasma membrane of neutrophils surface; intratracheal administration group, and capillary endothelial cells or alveolar epithelial cells in contact The activity of ALP on the surface of granulocyte plasma membrane was enhanced, while the ALP activity on the plasma membrane of the neutrophil in the alveolar cavity was weakened. In each group of specimens, the control group were AL P negative. Conclusion The ALP activity of rat lung PMN can be enhanced by LPS injection, and its activity may be related to the cell-cell adhesion, extracellular environment and the cell’s functional status.