Nomoto Y, Kuwano K, Hagimoto N, Kunitake R, Kawasaki M, Hara N
Research Institute for Diseases of the Chest, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Eur Respir J. 1997 Oct;10(10):2351-9. doi: 10.1183/09031936.97.10102351.
Deoxyribonucleic acid (DNA) strand breaks as a characteristic of apoptosis, and Fas antigen (Fas)/Fas ligand (FasL) expression may participate in acute immune complex alveolitis in mice. Male Institute for Cancer Research (ICR) mice were injected intravenously with immunoglobulin G (IgG) antibodies against ovalbumin and inhaled an aerosolized oval albumin (OA) solution. They were killed at 4, 6, 12, 24, 48 h and 7 days after aerosolization. We assessed DNA fragmentation by agarose gel electrophoresis and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate biotin nick end-labelling (TUNEL). The expression of Fas and FasL messenger ribonucleic acid (mRNA) in lung tissues was assessed by reverse transcriptase (RT) polymerase chain reaction, and by in situ hybridization (ISH) to localize Fas mRNA, and RT in situ polymerase chain reaction to localize FasL mRNA. The fragmentation of DNA extracted from lung tissue was found 6-24 h after OA inhalation. TUNEL detected positive signals in bronchial and alveolar epithelial, endothelial and inflammatory cells in the lung tissue. These positive signals had disappeared 7 days after OA inhalation. TUNEL also detected positive signals in apoptotic neutrophils in bronchoalveolar lavage fluid at 6-12 h. Fas mRNA was expressed in the alveolar epithelial and inflammatory cells, while the expression of FasL mRNA appeared to be upregulated in infiltrating inflammatory cells at 6-24 h. These results suggest that apoptosis may be associated with the resolution of inflammation and with tissue repair and also suggest the involvement of the Fas antigen/Fas ligand pathway in acute immune complex alveolitis in mice.
脱氧核糖核酸（DNA）链断裂是细胞凋亡的一个特征，而Fas抗原（Fas）/Fas配体（FasL）的表达可能参与小鼠急性免疫复合物性肺泡炎的发生。将雄性癌症研究所（ICR）小鼠静脉注射抗卵清蛋白的免疫球蛋白G（IgG）抗体，然后吸入雾化的卵清蛋白（OA）溶液。在雾化后4、6、12、24、48小时和7天处死小鼠。我们通过琼脂糖凝胶电泳和末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸生物素缺口末端标记法（TUNEL）评估DNA片段化情况。通过逆转录（RT）聚合酶链反应、原位杂交（ISH）定位Fas mRNA以及RT原位聚合酶链反应定位FasL mRNA来评估肺组织中Fas和FasL信使核糖核酸（mRNA）的表达。吸入OA后6 - 24小时发现从肺组织提取的DNA发生片段化。TUNEL在肺组织的支气管和肺泡上皮细胞以及内皮细胞和炎性细胞中检测到阳性信号。吸入OA 7天后这些阳性信号消失。TUNEL在支气管肺泡灌洗液中的凋亡中性粒细胞中于6 - 12小时也检测到阳性信号。Fas mRNA在肺泡上皮细胞和炎性细胞中表达，而FasL mRNA的表达在6 - 24小时似乎在浸润的炎性细胞中上调。这些结果表明细胞凋亡可能与炎症的消退和组织修复有关，也提示Fas抗原/Fas配体途径参与小鼠急性免疫复合物性肺泡炎的发生。
