Lung regeneration : implantation of fetal rat lung fragments into adult rat lung with bleomycine induced fibrosis

肺再生：将胎鼠肺碎片植入成年大鼠肺，博来霉素诱导纤维化

• 批准号: 17591474
• 负责人: SEKIYAMA Syoji
• 金额: $ 2.24万
• 依托单位: The University of Tokushima
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17591474/
• 关键词: lung regeneration; lung; rat; fetal lung tissue; fetal lung; transplantation; pulmonary fibrosis; bleomycine; 胎仔肺移植; 肺移植; ブレオマイシン; 再生; 組織修復

We previously reported that normal fetal lung tissues could survive and further grow and differentiate inside normal adult lung (J Thorac Cardiovasc Surg 2006; 131; 1148-53). In the present study, normal fetal lung tissues were implanted to bleomycine induced fibrotic adult rat lung parenchyma. GFP-LEW fetal lung tissues (day 17) were used as a donor. Normal adult rats were treated with intratracheal administration of bleomycine via a fine catheter to cause a situation of pulmonary fibrosis. A catheter (tracheotomy tube) was implanted in each rat three days before dosing. Bleomycine at a total dose of 4.5mg/kg or 3mg/kg was administered in four divided doses. Saline was administered for control rats.Day 17 fetal rat lungs tissues (GFP positive) in culture medium implanted to adult rats treated with bleomycine 3 or 4 weeks ago. To suppress immune responses to GFP antigen, recipient rats were administrated cyclosporine (10mg/kg i.m. on alternate days from day 0, dose of cyclosporine was decrease gradually). Recipient rats were killed 1, 2, 4, 8, 12 weeks after implantation. Recipient lungs implanted with fetal lung tissues were serially cut and examined under fluorescence microscope. The same specimens were stained with hematoxylin and eosin.The grafts implanted into pulmonary parenchyma with bleomycine induced fibrosis could survive and grow and differentiated with opening of the alveolar space.

我们之前报道过正常胎儿肺组织可以在正常成人肺内存活并进一步生长和分化（J Thorac cardiovascular Surg 2006; 131; 1148-53）。本研究将正常胎儿肺组织植入博来霉素诱导的纤维化成年大鼠肺实质。用GFP-LEW胎儿肺组织（第17天）作为供体。正常成年大鼠经细导管气管内给药博来霉素引起肺纤维化。给药前3天在每只大鼠体内植入一根导管（气管切开管）。博莱霉素总剂量为4.5mg/kg或3mg/kg，分4次给药。对照组大鼠给予生理盐水。第17天，胚胎大鼠肺组织（GFP阳性）在3或4周前植入博莱霉素处理的成年大鼠培养基中。为了抑制受体大鼠对GFP抗原的免疫应答，从第0天开始，隔天给药环孢素（10mg/kg），剂量逐渐减少。分别于植入后1、2、4、8、12周处死受体大鼠。连续切取胚胎肺组织植入受体肺，在荧光显微镜下观察。同样的标本用苏木精和伊红染色。植入博来霉素诱导纤维化的肺实质的移植物能够存活、生长和分化，肺泡间隙开放。