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脂多糖诱发急性肺炎时中性粒细胞碱性磷酸酶细胞化学定位观察 |
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r septum,while the IT-injection of LPS resulted in a high accumulation in the alveolar spaces mainly.The ALP activity was localized predominantly in small spherical and tubular membrane-bounded granules in neutrophils and observed occasionally in the non-inflamed regions of the rat lung.Following the IP-injected.Prominent ALP activity was found both on the plasma membrane surface and the cytoplasm of the neutrophils.In the IT-injected rat,the ALP activity was significantly increased on the membrane surface of the neutrophils contacting to the endothelial cells of the capillaries or the alveolar epithelial cells in the pneumonia region,while showed a weak reaction on the surface of the neutrophils which suspended in the alveolar spaces.No enzyme activity was detected at the endothelial plasma membranes in all of the samples.Negative reactions were showed in the controls. Conclusion ALP activity of neutrophils in the rat lung can be increased by LPS stimulation and its activity may be related to cell-cell interaction,the cell surroundings and the different functional status of the neutrophils.
Key words：Cytochemistry; Neutrophil; Alkaline phosphatase; Lipopolysaccharide; Rat lung.▲
长期以来，对被激活中性粒细胞质膜上的ALP活性意义不清。在Lopez和Yong的研究中［1］，曾以气管内滴注LPS刺激中性粒细胞大量聚于大鼠肺内，在支气管肺泡灌洗液中检出有高ALP活性。然而，一直未能对ALP精确定位，亦未对其增高的ALP活性意义作详尽的讨论。这里，我们利用一种实验性的急性炎性模型来研究大鼠肺内中性粒细胞ALP活性变化及其与中性粒细胞分布间的相互关系。
材料和方法
1. 试剂
购自Sigma Chemical Co.(St.Louis,MO,USA)的试剂：Lipopolysaccharide (LPS),p-nitrophenyl phosphate(p-NPP),N-tris［hydroxymethyl］ methylglycine(Tricine),N-tris［hydroxymetryl］ methyl-3-amino propane sulfonic acid (TAPS),saponin,Triton X-100,Levamisole;购自Nacalai Tesque Inc.(Kyoto,Japan)的试剂：Cerium chloride(CeCl3)，demethyl sulfoxide(DMSO);Osmium tetroxide(OsO4)购于PGM Chemicals(PTY)Ltd.(Germany,3620 R.S.A.);生物胶购自Sankyo Co.Ltd.(Tokyo,Japan)。
2. 动物处理
取体重250～350g的健康雄性SD系成年大白鼠(Japan SLC Inc.,Hamamatsu,Japan)24只，经LPS实验性诱发急性肺炎。分为腹腔内(intra-peritoneal,IP)给上一页 [1] [2] [3] 下一页
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